OBJECTIVE To examine the possibility of human sodium iodide symporter （hNIS） protein expression in lung cancer cells. METHODS Human lung A549 cancer cells were thawed and cultured in vitro. The cells were divided in...OBJECTIVE To examine the possibility of human sodium iodide symporter （hNIS） protein expression in lung cancer cells. METHODS Human lung A549 cancer cells were thawed and cultured in vitro. The cells were divided into an experimental group transfected with a recombinant pcDNA3-hNIS plasmid and a control group transfected only with a pcDNA3 plasmid. The recombinant plasmid vector encoding the hNIS gene （pcDNA3-hNIS） was amplified, purified and identified. The hNIS gene was followed by DNA sequencing. A Western blot and an immunohistochemical assay were applied to detect the hNIS protein expression in the transfected human lung A549 cancer cells. RESULTS Restriction enzyme digestion and DNA sequencing results showed the size and direction of the inserted gene in the recombinant pcD- NA3-hNIS plasmid was correct. The Western blot method and immunohistochemical analysis showed a positive NIS protein expression in the experimental group. The NIS protein was detected mainly in the cell membranes showing a positive rate up to 70.6% with no expression of the NIS protein in the control group. There was a significant difference between two groups （P=0.000）. CONCLUSION The hNIS gene was transfected effectively into human lung A549 cancer cells mediated by Lipofectamine 2000, and was expressed with its protein in vitro.展开更多
Radioiodine therapy,the most effective form of systemic radiotherapy available,is currently useful only for thyroid cancer because of the thyroid-specific expression of the human sodium iodide symporter(hNIS).Here,we ...Radioiodine therapy,the most effective form of systemic radiotherapy available,is currently useful only for thyroid cancer because of the thyroid-specific expression of the human sodium iodide symporter(hNIS).Here,we explore the efficacy of a novel form of gene therapy using prostate-specific membrane antigen(PSMA)promoter-mediated hNIS gene transfer followed by radioiodine administration for the treatment of castration-resistant prostate cancer(CRPC).The androgen-dependent C33 LNCaP cell line and the androgen-independent C81 LNCaP cell line were transfected by adenovirus.PSMA promoter-hNIS(Ad.PSMApro-hNIS)or adenovirus.cytomegalovirus-hNIS containing the cytomegalovirus promoter(Ad.CMM-hNIS)or a control virus.The iodide uptake was measured in vitro.The in vivo iodide uptake by C81 cell xenografts in nude mice injected with an adenovirus carrying the hNIS gene linked to PSMA and the corresponding tumor volume fluctuation were assessed.Iodide accumulation was shown in different LNCaP cell lines after Ad.PSMApro-hNIS and Ad.CMV-hNIS infection,but not in different LNCaP cell lines after adenovirus.cytomegalovirus(Ad.CMV)infection.At each time point,higher iodide uptake was shown in the C81 cells infected with Ad.PSMApro-hNIS than in the C33 cells(P〈0.05).An in vivo animal model showed a significant difference in 1311 radioiodine uptake in the tumors infected with Ad.PSMApro-hNIS,Ad.CMV-hNIS and control virus(P〈0.05)and a maximum reduction of tumor volume in mice infected with Ad.PSMApro-hNIS.These results show prostate-specific expression of the hNIS gene delivered by the PSMA promoter and effective radioiodine therapy of CRPC by the PSMA promoter-driven hNIS transfection.展开更多
Objective We investigated the correlation between the expression of the sodium-iodide symporter(NIS) and the detection of circulating tumor cells(CTCs) in differentiated thyroid carcinoma(DTC).Methods NIS expression i...Objective We investigated the correlation between the expression of the sodium-iodide symporter(NIS) and the detection of circulating tumor cells(CTCs) in differentiated thyroid carcinoma(DTC).Methods NIS expression in differentiated thyroid and the positive rate of CTCs in the peripheral blood were determined by immunohistochemistry S-P and flow cytometry from the records of 172 cases of differentiated thyroid carcinoma.Results Seventy-six cases(44.2%) expressed NIS in the differentiated thyroid and 63 cases(36.6%) were positive for CTCs in the peripheral blood. There was a significant difference between N0 and N1 in the expression of NIS(χ~2 = 6.015, P = 0.014) and the positive rate of CTCs(χ~2 = 14.035, P = 0.001). N0 and N1 also differed significantly in the expression of NIS(r =-0.383,-0.610, P = 0.002, < 0.001). The differences in the NIS expression, but not in the positive rate of CTCs, were significant among the different pathological subtypes(χ~2 = 7.897, P = 0.005; χ~2 = 1.455, P = 0.228, respectively). There was a significant negative correlation between the highly differentiated type and intermediate differentiation type both in the expression of NIS and positive rate of CTCs(r =-0.591,-0.443, P < 0.001, P = 0.002). Conclusion There was a significant negative correlation between the expression of tissue NIS and positive rate of CTCs in the peripheral blood in DTC. The malignancy level and lymph node metastasis in differentiated thyroid carcinoma were negatively correlated with NIS expression and positively correlated with the positive rate of CTC.展开更多
Objective: To investigate the effect of cytokines (TNF-α,IFN-γ and IL-6) on the expression of sodium-iodide symporter(NIS)gene in breast cancer cell(MCF-7). Methods:The breast cancer cell was cultureds with negativ...Objective: To investigate the effect of cytokines (TNF-α,IFN-γ and IL-6) on the expression of sodium-iodide symporter(NIS)gene in breast cancer cell(MCF-7). Methods:The breast cancer cell was cultureds with negative control culture or culture with different concentrations of cytokines for 72 h.NIS gene mRNA in breast cancer cells cultured was determined by reverse transcriptase-polymerase chain reaction(RT-PCR). Results:Expression of sodium-iodide symporter mRNA can be found decreasing along with increasing the concentration of cytokine dose-dependently. Conclusion: Cytokine may play a role in iodide-uptake modulating in breast cancer cells by their effect on NIS gene expression.展开更多
Objective; The aim of this study was to investigate the effects of radiofrequency treatment on sodium/iodide symporter expression of thyroid cancer ceils. Methods： In 29 thyroid cancer patients with low or no express...Objective; The aim of this study was to investigate the effects of radiofrequency treatment on sodium/iodide symporter expression of thyroid cancer ceils. Methods： In 29 thyroid cancer patients with low or no expression of soda ／ iodide symporter, the radio frequency combined 1311 therapy was used, the whole-body scintigraphy and serum Ig were detected before and after the radiofrequency treatment. Results： The whole-body scintigraphy showed that 4 cases （4/29） before radiofrequenc_y treatment had positive iodine uptake, 19 cases （19/29） two weeks after radiofrequency treatment had the positive iodine uptake, 12 cases （12/29） four weeks after radiofrequency treatment had the positive iodine uptake. Four weeks after radiofrequency treatment, 5 cases had increased serum Ig levels, 17 cases had decreased serum Ig levels, 7 cases showed no change. 25 cases （25/29） were effective, 15 cases （15/29） were cured. Conclusion： The radiofrequency induced the non-expressed the sodium/iodide symporter of thyroid cancer cells regain the iodine intake ability, it improved the clinical efficacy of 131I therapy in dedifferentiated thyroid cancer.展开更多
Objective： The aim of our study was to investigate the correlation between the expression of sodium/iodide symporter, serum levels of β2-MG and prognosis of thyroid carcinoma patients. Methods： Ninty-five cases wit...Objective： The aim of our study was to investigate the correlation between the expression of sodium/iodide symporter, serum levels of β2-MG and prognosis of thyroid carcinoma patients. Methods： Ninty-five cases with thyroid carcinoma, using enzyme-linked immunosorbent assay with double-antibody sandwich to detect the serum β2-MG levels and immunehistochemistry to detect NIS expression of thyroid cancer tissue. Results： Thirty-seven cases showed positive expression of sodium/iodide symporter （38.9%） and 30 cases showed positive expression of β2-MG （31.57%）. There were significant differences of NIS expression （X2 = 8.207, P = 0.017） and β2-MG expression （X2 = 10.121, P = 0.006） between different pathological types of thyroid carcinoma, but there was no correlation between the positive rate of the two research groups （r = -0.546, P = 0.633）. The significant differences was observed in expression of sodium/iodide symporter （X2 = 9.272, P=0.002） and expression of β2-MG （X2 = 4.441, P = 0.035） between the group with neck lymph node metastasis and the group without neck lymph node metastasis and both positive rate was significantly negatively correlated （r = -1.000, P = 0.000）. The significant differences was observed in expression of sodium/iodide symporter （X2 = 9.272, P = 0.002） and expression of β2-MG （X2 = 3.867, P = 0.043） between the group with distant organ metastasis and the group without distant organ metastasis （X2 = 11.985, P = 0.001） and both positive rate was significantly negatively correlated （r = -1.000, P = 0.000）. Conclusion： There are significantly negatively correlated between neck lymph node metastasis, distant organ metastasis and expression of sodium/iodide symporter and expression of β2-MG. Thyroid cancer lymph node and distant organ metastasis, the tumor tissue NIS expression and serum levels of β2-MG is significantly negatively correlated. The detection of serum β2-MG provides clinical reference value for the effects on radionuclide therapy and prognosis assessment of thyroid carcinoma. Serum β2-MG levels is negatively correlated with prognosis in thyroid cancer patients.展开更多
Radioiodine ablation（RIA） therapy is one of the most important treatments for papillary thyroid carcinoma（PTC）, but some patients who received 131 I have radioiodine-refractory disease caused by the decreased expr...Radioiodine ablation（RIA） therapy is one of the most important treatments for papillary thyroid carcinoma（PTC）, but some patients who received 131 I have radioiodine-refractory disease caused by the decreased expression of the Na^＋/I^- symporter（NIS）. BRAF^V600E mutation is one possible risk factor that can disturb the NIS expression, but the roles are unclear in clinical practice. This research discussed the association of BRAF^V600E mutation and NIS expression in PTC tissue and the clinical implications in RIA therapy. 134 PTC samples were collected between June 2013 and June 2014 from Tongji Hospital affiliated to Tongji Medical College, and their clinical characteristics were analyzed. RT-PCR was used to detect the BRAF^V600E mutation from formalin-fixed paraffin-embedded samples, and immunohistochemistry was applied to detect the NIS expression. IPP software was used to calculate the relative expression quantity of NIS. We found that there was no significant correlation between the absorbance（A） values of NIS and clinicopathologic features in these cases, even thyroid stimulating hormone. BRAF^V600E mutation showed inhibitory effect on the NIS expression without statistically significant difference in all PTC cases（β=–0.0195, P=0.085）, but in the subgroup without hashimoto＇s thyroiditis（HT）, BRAF^V600E mutation could significantly inhibit the NIS expression（β=–0.0257, P=0.046）. The results indicate that BRAF^V600E mutation is correlated with a lower expression of NIS in PTCs without HT, suggesting the radioiodine-refractory effects during RIA therapy in these patients.展开更多
To obtain human sodium/iodide symporter gene cDNA for studying its potential ability as a radioiodide treatment for melanoma, the hNIS gene cDNA was amplified with total RNA from human thyroid tissue by RT-PCR. The hN...To obtain human sodium/iodide symporter gene cDNA for studying its potential ability as a radioiodide treatment for melanoma, the hNIS gene cDNA was amplified with total RNA from human thyroid tissue by RT-PCR. The hNIS cDNA was inserted into cloning vector pUCm-T and subcloned into eukaryotic expression vector pc-DNA3. The pc-DNA3-hNIS and pc-DNA3 were transduced into melanoma cells (B16) by electroporation, and two cell lines termed B16-A and B16-B respectively were established. The uptake and efflux of iodide was examined in vitro. The three cell lines (B16-A, B16-B, B16) were injected subcutaneously into the right flank of C57 mice. Biodistribution study and tumor imaging were performed when the tumor reached approximately 10mm in diameter. The cloned hNIS cDNA sequence was identical with the published sequence. Two novel cell lines named 16-A containing pc-DNA3-hNIS and B16-B containing pc-DNA3 only were established. The resultant cell line B16-A accumulated 17 and 19 times more radioiodide in vitro than B16 and B16-B respectively. The iodide uptake reached the half-maximal level within 10 min, and reached a plateau at 30 min. The efflux of iodide was also rapid (T1/2eff=10min). The imaging shows in vivo uptake in expected sites including the salivary glands, thyroid, stomach, and hNIS-transduced tumor, whereas the nontransduced tumor was not visualized. The %ID/g of B16-A tumors at 1, 2, 4, 12, and 24h after injec- tion of 125I were 12.22±0.71, 10.91±0.72, 8.73±0.99, 1.24±0.29, and 0.19±0.03, respectively, which were signifi- cantly higher percentages than those for controlling tumors, p<0.01. However, biologic T1/2 was about 6 h. Our pre- liminary data indicate that the transduction of the hNIS gene per se is sufficient to induce iodide transport in mela- noma cells both in vitro and in vivo, but T1/2eff is short.展开更多
To investigate human sodium/iodide symporter (hNIS) induced iodine uptake in human lung adenocarcinoma via baculovirus, a recombinant baculovirus encoding hNIS gene was constructed under the control of CMV promoter (B...To investigate human sodium/iodide symporter (hNIS) induced iodine uptake in human lung adenocarcinoma via baculovirus, a recombinant baculovirus encoding hNIS gene was constructed under the control of CMV promoter (Bac-CMV-hNIS). In vitro, baculovirus infected A549 cells accumulated about 27 times more 125I than that of noninfected cells. The 125I uptake was maximal after 30-min incubation of the cells, and efflux of the radioactivity was rapid, with 50% lost during the first 2 min after 125I-containing medium had been replaced by nonradioactive medium. Competition experiments in the presence of sodium perchlorate revealed a dose-dependent decrease of 125I uptake. Bac-CMV-hNIS infected tumor cells were selectively killed by exposure to 131I, as revealed by clonogenic assays. In nude mice, Bac-CMV-hNIS infected A549 cells accumulated more 131I than that of the control monitored by 1-h scintigraphy after 131I administration. The transduction of hNIS gene through baculovirus is sufficient to induce iodine transporting in A549 cells in vitro and in vivo, outlining the potential of this novel tumor gene imaging approach. But a rapid efflux of radioactivity from the tumor was shown in vivo and the in vivo therapy test showed no sign of effect.展开更多
Background The sodium-iodide symporter （NIS） protein can mediate the active radioiodine uptake.The human telomerase reverse transcriptase （hTERT） promoter is known to be selectively reactivated in majority of tumo...Background The sodium-iodide symporter （NIS） protein can mediate the active radioiodine uptake.The human telomerase reverse transcriptase （hTERT） promoter is known to be selectively reactivated in majority of tumors and hence could be used for tumor targeting.We constructed a recombinant adenovirus containing the human sodium iodide symporter （hNIS） gene directed by the hTERT promoter, characterized the ability of infected cells in uptaking iodide, and explored the therapeutic efficacy of 131I in a lung cancer cell line in vitro.Methods The hTERT promoter was amplified by PCR from DNA isolated from log-phase HepG2 cells, subcloned into lineralized FL＊-hNIS/pcDNA3, and then the hTERT-hNIS sequence was subcloned into the shuttle plasmid pAdTrack.The recombinant adenovirus Ad-hTERT-hNIS was constructed by AdEasy system.A positive control adenovirusAd-CMV-hNIS and a negative control adenovirus Ad-CMV were created similarly.A549 cells were transduced with recombinant adenoviruses.125I uptake studies and sodium perchlorate suppression studies were used to confirm hNIS expression and function.Toxic effects of 131I on tumor cells were studied by in vitro clonogenic assay.Results We first successfully constructed an adenovirus mediated transgene expression system of the hNIS under the control of hTERT promoter.When infected with recombinant adenovirus constructs expressing hNIS directed by hTERTand CMV-promoters （Ad-hTERT-hNIS and Ad-CMV-hNIS, respectively）, the lung cancer cell line A549 had increased ability to uptake radioiodide up to 23- and 30- fold compared to the control parental cells, respectively.The radioiodide uptake ability of both the Ad-CMV-hNIS and Ad-hTERT-hNIS transduced cell lines were repressed 11-fold by sodium perchlorate （NaCIO4）.The subsequent in vitro clonogenic assay of the infected A549 cell line was further repressed to 23% （Ad-CMV-hNIS） and 30% （Ad-hTERT-hNIS） of the control group after receiving radioiodide for 7 hours （P 〈0.001）.Conclusion Our preliminary study indicates that an adenovirus mediated transgene expression system of the hNIS under the control of hTERT promoter has the potential to become an effective wide-spectrum yet highly specific anti-cancer strategy.展开更多
Anaplastic thyroid cancer(ATC)is a rare but highly lethal disease.ATCs are resistant to standard therapies and are extremely difficult to manage.The stepwise cell dedifferentiation results in the impairment of the iod...Anaplastic thyroid cancer(ATC)is a rare but highly lethal disease.ATCs are resistant to standard therapies and are extremely difficult to manage.The stepwise cell dedifferentiation results in the impairment of the iodine-metabolizing machinery and the infeasibility of radioiodine treatment in ATC.Hence,reinducing iodine-metabolizing gene expression to restore radioiodine avidity is considered as a promising strategy to fight against ATC.In the present study,capsaicin(CAP),a natural potent transient receptor potential vanilloid type 1(TRPV1)agonist,was discovered to reinduce ATC cell differentiation and to increase the expression of thyroid transcription factors(TTFs including TTF-1,TTF-2,and PAX8)and iodine-metabolizing proteins,including thyroidstimulating hormone receptor(TSHR),thyroid peroxidase,and sodium iodine symporter(NIS),in two ATC cell lines,8505C and FRO.Strikingly,CAP treatment promoted NIS glycosylation and its membrane trafficking,resulting in a significant enhancement of radioiodine uptake of ATC cells in vitro.Mechanistically,CAP-activated TRPV1 channel and subsequently triggered Ca2þinflux,cyclic adenosine monophosphate(cAMP)generation,and cAMP-responsive element-binding protein(CREB)signal activation.Next,CREB recognized and bound to the promoter of SLC5A5 to facilitate its transcription.Moreover,the TRPV1 antagonist CPZ,the calcium chelator BAPTA,and the PKA inhibitor H-89 effectively alleviated the redifferentiation exerted by CAP,demonstrating that CAP might improve radioiodine avidity through the activation of the TRPV1–Ca2þ/cAMP/PKA/CREB signaling pathway.In addition,our study indicated that CAP might trigger a novel cascade to redifferentiate ATC cells and provide unprecedented opportunities for radioiodine therapy in ATC,bypassing canonical TSH–TSHR pathway.展开更多
Synthetic molecules that can mediate the coupled transport of Cl^(-) with K^(+) and/or Na+across the lipid bilayers have aroused great interest due to their potential as a novel therapeutic strategy by disrupting cell...Synthetic molecules that can mediate the coupled transport of Cl^(-) with K^(+) and/or Na+across the lipid bilayers have aroused great interest due to their potential as a novel therapeutic strategy by disrupting cellular ion homeostasis.Based on the structural advantages of molecular rotaxanes,we herein show that two rotaxane-based transporters[2]R and[3]R induce coupled K^(+)/Cl^(-) channel transport by introducing Cl^(-) recognition sites in the thread and K^(+) binding group in the wheel,respectively.The welldesigned molecular structures allow the insertion of unimolecular rotaxanes into the lipid bilayer,thus achieving effective ion transport by means of thermodynamically controlled movement and driven by the difference in ion concentration inside and outside the vesicles.In addition,the use of a three-component rotaxane can accelerate ion transport through a cooperative shuttlerelay mechanism in which two wheels move along the thread in the lipid membrane,thereby enabling[3]R to have higher ion transport capacity.This work represents a major advance in the use of rotaxane molecules to accomplish more complex and effective tasks.展开更多
Objective The present study investigated the sodium/iodide symporter mRNA expression in mouse lactating mammary gland cells under different iodine levels and the effects of thyroid-stimulating hormone(TSH),estradiol(E...Objective The present study investigated the sodium/iodide symporter mRNA expression in mouse lactating mammary gland cells under different iodine levels and the effects of thyroid-stimulating hormone(TSH),estradiol(E2)and prolactin(PRL)on NIS mRNA expression in mouse lactating mammary gland cells.展开更多
Radioactive iodine(RAI)therapy remains a cornerstone in the management of differentiated thyroid cancer(DTC).The therapeutic efficacy of RAI depends on thyroid-stimulating hormone(TSH)-driven uptake via the sodium-iod...Radioactive iodine(RAI)therapy remains a cornerstone in the management of differentiated thyroid cancer(DTC).The therapeutic efficacy of RAI depends on thyroid-stimulating hormone(TSH)-driven uptake via the sodium-iodide symporter(NIS).Although elevated TSH levels are routinely achieved before RAI administration,the circadian rhythm of TSH,which peaks overnight,is largely overlooked in treatment protocols.Aligning RAI administration with this physiological peak,particularly through evening dosing,may enhance iodine uptake,improve therapeutic efficacy,and reduce systemic exposure.Preclinical data and the known circadian regulation of TSH and NIS support this approach,although clinical validation is lacking.Evening dosing may also improve patient convenience and clinic workflow.However,logistical challenges and the absence of human trials present barriers to implementation.In this narrative review,we synthesize current evidence and tentatively propose the chronotherapeutic alignment of RAI timing as a novel,potentially impactful optimization strategy in DTC management.Prospective studies are needed to evaluate its clinical benefits and feasibility.展开更多
The symbiotic association between reef-building corals and Symbiodiniaceae is pivotal for coral reef ecosystems,yet remains susceptible to environmental factors.Currently,there is a dearth of research examining season...The symbiotic association between reef-building corals and Symbiodiniaceae is pivotal for coral reef ecosystems,yet remains susceptible to environmental factors.Currently,there is a dearth of research examining seasonal fluctuations in coral-associated Symbiodiniaceae communities.In this study,we investigated the seasonal dynamics of Symbiodiniaceae communities associated with coral species in the Luhuitou coral reef using high-throughput sequencing techniques and SymPortal analytical framework.The results indicated that the genus Cladocopium exhibited dominance(averaging 82%),followed by Durusdinium(18%)and Breviolum(0.01%)within the examined coral species.Among the 521 Symbiodiniaceae ITS2 sequence types,C15 emerged as the prevalent type(13.24%),trailed by C3u(9.51%)and D1(8.57%).Interestingly,Symbiodiniaceae communities varied among different coral species.Pocillopora damicornis displayed a predominant association with Durusdinium,while Porites lutea,Goniastrea retiformis,Montipora truncata,Montipora aequituberculata,and Acropora divaricata were entirely dominated by the genus Cladocopium(100%),showcasing distinct host specificity.In the cases of Hydnophora exesa,Acropora latistella,Acropora digitifera,and seawater,both Cladocopium and Durusdinium were concurrently detected.Moreover,the diversity of Symbiodiniaceae associated with P.damicornis,P.lutea,G.retiformis,M.truncata,M.aequituberculata,and A.digitifera exhibited significant variations across different seasons.Notably,the results revealed that the alterations in Symbiodiniaceae community compositions were primarily driven by nutrient concentrations and seawater temperature.The network analysis of Symbiodiniaceae revealed the dominant Symbiodiniaceae types C15,C17f,C3u,C3,and D4 were exclusive.This study provided the seasonal variation characteristics of Symbiodiniaceae communities among different coral species,which may be a potential adaptive mechanism to environmental conditions.展开更多
文摘OBJECTIVE To examine the possibility of human sodium iodide symporter （hNIS） protein expression in lung cancer cells. METHODS Human lung A549 cancer cells were thawed and cultured in vitro. The cells were divided into an experimental group transfected with a recombinant pcDNA3-hNIS plasmid and a control group transfected only with a pcDNA3 plasmid. The recombinant plasmid vector encoding the hNIS gene （pcDNA3-hNIS） was amplified, purified and identified. The hNIS gene was followed by DNA sequencing. A Western blot and an immunohistochemical assay were applied to detect the hNIS protein expression in the transfected human lung A549 cancer cells. RESULTS Restriction enzyme digestion and DNA sequencing results showed the size and direction of the inserted gene in the recombinant pcD- NA3-hNIS plasmid was correct. The Western blot method and immunohistochemical analysis showed a positive NIS protein expression in the experimental group. The NIS protein was detected mainly in the cell membranes showing a positive rate up to 70.6% with no expression of the NIS protein in the control group. There was a significant difference between two groups （P=0.000）. CONCLUSION The hNIS gene was transfected effectively into human lung A549 cancer cells mediated by Lipofectamine 2000, and was expressed with its protein in vitro.
文摘Radioiodine therapy,the most effective form of systemic radiotherapy available,is currently useful only for thyroid cancer because of the thyroid-specific expression of the human sodium iodide symporter(hNIS).Here,we explore the efficacy of a novel form of gene therapy using prostate-specific membrane antigen(PSMA)promoter-mediated hNIS gene transfer followed by radioiodine administration for the treatment of castration-resistant prostate cancer(CRPC).The androgen-dependent C33 LNCaP cell line and the androgen-independent C81 LNCaP cell line were transfected by adenovirus.PSMA promoter-hNIS(Ad.PSMApro-hNIS)or adenovirus.cytomegalovirus-hNIS containing the cytomegalovirus promoter(Ad.CMM-hNIS)or a control virus.The iodide uptake was measured in vitro.The in vivo iodide uptake by C81 cell xenografts in nude mice injected with an adenovirus carrying the hNIS gene linked to PSMA and the corresponding tumor volume fluctuation were assessed.Iodide accumulation was shown in different LNCaP cell lines after Ad.PSMApro-hNIS and Ad.CMV-hNIS infection,but not in different LNCaP cell lines after adenovirus.cytomegalovirus(Ad.CMV)infection.At each time point,higher iodide uptake was shown in the C81 cells infected with Ad.PSMApro-hNIS than in the C33 cells(P〈0.05).An in vivo animal model showed a significant difference in 1311 radioiodine uptake in the tumors infected with Ad.PSMApro-hNIS,Ad.CMV-hNIS and control virus(P〈0.05)and a maximum reduction of tumor volume in mice infected with Ad.PSMApro-hNIS.These results show prostate-specific expression of the hNIS gene delivered by the PSMA promoter and effective radioiodine therapy of CRPC by the PSMA promoter-driven hNIS transfection.
基金Supported by a grant from the Gansu Province Key Traditional Chinese Medicine Project(No.GZK-2010-Z9)
文摘Objective We investigated the correlation between the expression of the sodium-iodide symporter(NIS) and the detection of circulating tumor cells(CTCs) in differentiated thyroid carcinoma(DTC).Methods NIS expression in differentiated thyroid and the positive rate of CTCs in the peripheral blood were determined by immunohistochemistry S-P and flow cytometry from the records of 172 cases of differentiated thyroid carcinoma.Results Seventy-six cases(44.2%) expressed NIS in the differentiated thyroid and 63 cases(36.6%) were positive for CTCs in the peripheral blood. There was a significant difference between N0 and N1 in the expression of NIS(χ~2 = 6.015, P = 0.014) and the positive rate of CTCs(χ~2 = 14.035, P = 0.001). N0 and N1 also differed significantly in the expression of NIS(r =-0.383,-0.610, P = 0.002, < 0.001). The differences in the NIS expression, but not in the positive rate of CTCs, were significant among the different pathological subtypes(χ~2 = 7.897, P = 0.005; χ~2 = 1.455, P = 0.228, respectively). There was a significant negative correlation between the highly differentiated type and intermediate differentiation type both in the expression of NIS and positive rate of CTCs(r =-0.591,-0.443, P < 0.001, P = 0.002). Conclusion There was a significant negative correlation between the expression of tissue NIS and positive rate of CTCs in the peripheral blood in DTC. The malignancy level and lymph node metastasis in differentiated thyroid carcinoma were negatively correlated with NIS expression and positively correlated with the positive rate of CTC.
文摘Objective: To investigate the effect of cytokines (TNF-α,IFN-γ and IL-6) on the expression of sodium-iodide symporter(NIS)gene in breast cancer cell(MCF-7). Methods:The breast cancer cell was cultureds with negative control culture or culture with different concentrations of cytokines for 72 h.NIS gene mRNA in breast cancer cells cultured was determined by reverse transcriptase-polymerase chain reaction(RT-PCR). Results:Expression of sodium-iodide symporter mRNA can be found decreasing along with increasing the concentration of cytokine dose-dependently. Conclusion: Cytokine may play a role in iodide-uptake modulating in breast cancer cells by their effect on NIS gene expression.
文摘Objective; The aim of this study was to investigate the effects of radiofrequency treatment on sodium/iodide symporter expression of thyroid cancer ceils. Methods： In 29 thyroid cancer patients with low or no expression of soda ／ iodide symporter, the radio frequency combined 1311 therapy was used, the whole-body scintigraphy and serum Ig were detected before and after the radiofrequency treatment. Results： The whole-body scintigraphy showed that 4 cases （4/29） before radiofrequenc_y treatment had positive iodine uptake, 19 cases （19/29） two weeks after radiofrequency treatment had the positive iodine uptake, 12 cases （12/29） four weeks after radiofrequency treatment had the positive iodine uptake. Four weeks after radiofrequency treatment, 5 cases had increased serum Ig levels, 17 cases had decreased serum Ig levels, 7 cases showed no change. 25 cases （25/29） were effective, 15 cases （15/29） were cured. Conclusion： The radiofrequency induced the non-expressed the sodium/iodide symporter of thyroid cancer cells regain the iodine intake ability, it improved the clinical efficacy of 131I therapy in dedifferentiated thyroid cancer.
文摘Objective： The aim of our study was to investigate the correlation between the expression of sodium/iodide symporter, serum levels of β2-MG and prognosis of thyroid carcinoma patients. Methods： Ninty-five cases with thyroid carcinoma, using enzyme-linked immunosorbent assay with double-antibody sandwich to detect the serum β2-MG levels and immunehistochemistry to detect NIS expression of thyroid cancer tissue. Results： Thirty-seven cases showed positive expression of sodium/iodide symporter （38.9%） and 30 cases showed positive expression of β2-MG （31.57%）. There were significant differences of NIS expression （X2 = 8.207, P = 0.017） and β2-MG expression （X2 = 10.121, P = 0.006） between different pathological types of thyroid carcinoma, but there was no correlation between the positive rate of the two research groups （r = -0.546, P = 0.633）. The significant differences was observed in expression of sodium/iodide symporter （X2 = 9.272, P=0.002） and expression of β2-MG （X2 = 4.441, P = 0.035） between the group with neck lymph node metastasis and the group without neck lymph node metastasis and both positive rate was significantly negatively correlated （r = -1.000, P = 0.000）. The significant differences was observed in expression of sodium/iodide symporter （X2 = 9.272, P = 0.002） and expression of β2-MG （X2 = 3.867, P = 0.043） between the group with distant organ metastasis and the group without distant organ metastasis （X2 = 11.985, P = 0.001） and both positive rate was significantly negatively correlated （r = -1.000, P = 0.000）. Conclusion： There are significantly negatively correlated between neck lymph node metastasis, distant organ metastasis and expression of sodium/iodide symporter and expression of β2-MG. Thyroid cancer lymph node and distant organ metastasis, the tumor tissue NIS expression and serum levels of β2-MG is significantly negatively correlated. The detection of serum β2-MG provides clinical reference value for the effects on radionuclide therapy and prognosis assessment of thyroid carcinoma. Serum β2-MG levels is negatively correlated with prognosis in thyroid cancer patients.
基金supported by Hubei Natural Science Foundation of China(No.2014CKB519)
文摘Radioiodine ablation（RIA） therapy is one of the most important treatments for papillary thyroid carcinoma（PTC）, but some patients who received 131 I have radioiodine-refractory disease caused by the decreased expression of the Na^＋/I^- symporter（NIS）. BRAF^V600E mutation is one possible risk factor that can disturb the NIS expression, but the roles are unclear in clinical practice. This research discussed the association of BRAF^V600E mutation and NIS expression in PTC tissue and the clinical implications in RIA therapy. 134 PTC samples were collected between June 2013 and June 2014 from Tongji Hospital affiliated to Tongji Medical College, and their clinical characteristics were analyzed. RT-PCR was used to detect the BRAF^V600E mutation from formalin-fixed paraffin-embedded samples, and immunohistochemistry was applied to detect the NIS expression. IPP software was used to calculate the relative expression quantity of NIS. We found that there was no significant correlation between the absorbance（A） values of NIS and clinicopathologic features in these cases, even thyroid stimulating hormone. BRAF^V600E mutation showed inhibitory effect on the NIS expression without statistically significant difference in all PTC cases（β=–0.0195, P=0.085）, but in the subgroup without hashimoto＇s thyroiditis（HT）, BRAF^V600E mutation could significantly inhibit the NIS expression（β=–0.0257, P=0.046）. The results indicate that BRAF^V600E mutation is correlated with a lower expression of NIS in PTCs without HT, suggesting the radioiodine-refractory effects during RIA therapy in these patients.
文摘To obtain human sodium/iodide symporter gene cDNA for studying its potential ability as a radioiodide treatment for melanoma, the hNIS gene cDNA was amplified with total RNA from human thyroid tissue by RT-PCR. The hNIS cDNA was inserted into cloning vector pUCm-T and subcloned into eukaryotic expression vector pc-DNA3. The pc-DNA3-hNIS and pc-DNA3 were transduced into melanoma cells (B16) by electroporation, and two cell lines termed B16-A and B16-B respectively were established. The uptake and efflux of iodide was examined in vitro. The three cell lines (B16-A, B16-B, B16) were injected subcutaneously into the right flank of C57 mice. Biodistribution study and tumor imaging were performed when the tumor reached approximately 10mm in diameter. The cloned hNIS cDNA sequence was identical with the published sequence. Two novel cell lines named 16-A containing pc-DNA3-hNIS and B16-B containing pc-DNA3 only were established. The resultant cell line B16-A accumulated 17 and 19 times more radioiodide in vitro than B16 and B16-B respectively. The iodide uptake reached the half-maximal level within 10 min, and reached a plateau at 30 min. The efflux of iodide was also rapid (T1/2eff=10min). The imaging shows in vivo uptake in expected sites including the salivary glands, thyroid, stomach, and hNIS-transduced tumor, whereas the nontransduced tumor was not visualized. The %ID/g of B16-A tumors at 1, 2, 4, 12, and 24h after injec- tion of 125I were 12.22±0.71, 10.91±0.72, 8.73±0.99, 1.24±0.29, and 0.19±0.03, respectively, which were signifi- cantly higher percentages than those for controlling tumors, p<0.01. However, biologic T1/2 was about 6 h. Our pre- liminary data indicate that the transduction of the hNIS gene per se is sufficient to induce iodide transport in mela- noma cells both in vitro and in vivo, but T1/2eff is short.
基金Supported by National Natural Science Foundation of China (No.30900375)Shanghai Leading Academic Discipline Project (No.S30203)Medical Engineering (Science) Cross micro PET Special Foundation of Shanghai Jiaotong University (No.YG08PETZD01)
文摘To investigate human sodium/iodide symporter (hNIS) induced iodine uptake in human lung adenocarcinoma via baculovirus, a recombinant baculovirus encoding hNIS gene was constructed under the control of CMV promoter (Bac-CMV-hNIS). In vitro, baculovirus infected A549 cells accumulated about 27 times more 125I than that of noninfected cells. The 125I uptake was maximal after 30-min incubation of the cells, and efflux of the radioactivity was rapid, with 50% lost during the first 2 min after 125I-containing medium had been replaced by nonradioactive medium. Competition experiments in the presence of sodium perchlorate revealed a dose-dependent decrease of 125I uptake. Bac-CMV-hNIS infected tumor cells were selectively killed by exposure to 131I, as revealed by clonogenic assays. In nude mice, Bac-CMV-hNIS infected A549 cells accumulated more 131I than that of the control monitored by 1-h scintigraphy after 131I administration. The transduction of hNIS gene through baculovirus is sufficient to induce iodine transporting in A549 cells in vitro and in vivo, outlining the potential of this novel tumor gene imaging approach. But a rapid efflux of radioactivity from the tumor was shown in vivo and the in vivo therapy test showed no sign of effect.
基金This work was supported by Project of Natural Science Foundation of Jiangsu Province （NO. BK2008164）.
文摘Background The sodium-iodide symporter （NIS） protein can mediate the active radioiodine uptake.The human telomerase reverse transcriptase （hTERT） promoter is known to be selectively reactivated in majority of tumors and hence could be used for tumor targeting.We constructed a recombinant adenovirus containing the human sodium iodide symporter （hNIS） gene directed by the hTERT promoter, characterized the ability of infected cells in uptaking iodide, and explored the therapeutic efficacy of 131I in a lung cancer cell line in vitro.Methods The hTERT promoter was amplified by PCR from DNA isolated from log-phase HepG2 cells, subcloned into lineralized FL＊-hNIS/pcDNA3, and then the hTERT-hNIS sequence was subcloned into the shuttle plasmid pAdTrack.The recombinant adenovirus Ad-hTERT-hNIS was constructed by AdEasy system.A positive control adenovirusAd-CMV-hNIS and a negative control adenovirus Ad-CMV were created similarly.A549 cells were transduced with recombinant adenoviruses.125I uptake studies and sodium perchlorate suppression studies were used to confirm hNIS expression and function.Toxic effects of 131I on tumor cells were studied by in vitro clonogenic assay.Results We first successfully constructed an adenovirus mediated transgene expression system of the hNIS under the control of hTERT promoter.When infected with recombinant adenovirus constructs expressing hNIS directed by hTERTand CMV-promoters （Ad-hTERT-hNIS and Ad-CMV-hNIS, respectively）, the lung cancer cell line A549 had increased ability to uptake radioiodide up to 23- and 30- fold compared to the control parental cells, respectively.The radioiodide uptake ability of both the Ad-CMV-hNIS and Ad-hTERT-hNIS transduced cell lines were repressed 11-fold by sodium perchlorate （NaCIO4）.The subsequent in vitro clonogenic assay of the infected A549 cell line was further repressed to 23% （Ad-CMV-hNIS） and 30% （Ad-hTERT-hNIS） of the control group after receiving radioiodide for 7 hours （P 〈0.001）.Conclusion Our preliminary study indicates that an adenovirus mediated transgene expression system of the hNIS under the control of hTERT promoter has the potential to become an effective wide-spectrum yet highly specific anti-cancer strategy.
基金supported by grants from the National Natural Science Foundation of China(81972503 and 82103656).
文摘Anaplastic thyroid cancer(ATC)is a rare but highly lethal disease.ATCs are resistant to standard therapies and are extremely difficult to manage.The stepwise cell dedifferentiation results in the impairment of the iodine-metabolizing machinery and the infeasibility of radioiodine treatment in ATC.Hence,reinducing iodine-metabolizing gene expression to restore radioiodine avidity is considered as a promising strategy to fight against ATC.In the present study,capsaicin(CAP),a natural potent transient receptor potential vanilloid type 1(TRPV1)agonist,was discovered to reinduce ATC cell differentiation and to increase the expression of thyroid transcription factors(TTFs including TTF-1,TTF-2,and PAX8)and iodine-metabolizing proteins,including thyroidstimulating hormone receptor(TSHR),thyroid peroxidase,and sodium iodine symporter(NIS),in two ATC cell lines,8505C and FRO.Strikingly,CAP treatment promoted NIS glycosylation and its membrane trafficking,resulting in a significant enhancement of radioiodine uptake of ATC cells in vitro.Mechanistically,CAP-activated TRPV1 channel and subsequently triggered Ca2þinflux,cyclic adenosine monophosphate(cAMP)generation,and cAMP-responsive element-binding protein(CREB)signal activation.Next,CREB recognized and bound to the promoter of SLC5A5 to facilitate its transcription.Moreover,the TRPV1 antagonist CPZ,the calcium chelator BAPTA,and the PKA inhibitor H-89 effectively alleviated the redifferentiation exerted by CAP,demonstrating that CAP might improve radioiodine avidity through the activation of the TRPV1–Ca2þ/cAMP/PKA/CREB signaling pathway.In addition,our study indicated that CAP might trigger a novel cascade to redifferentiate ATC cells and provide unprecedented opportunities for radioiodine therapy in ATC,bypassing canonical TSH–TSHR pathway.
基金supported by the National Natural Science Foundation of China(22171085)the Shanghai Science Technology Communication(21ZR1415500)Shanghai Frontier Science Research Base of Optogenetic Techniques for Cell Metabolism(Shanghai Municipal Education Commission,Grant 2021 Sci&Tech 03-28)。
文摘Synthetic molecules that can mediate the coupled transport of Cl^(-) with K^(+) and/or Na+across the lipid bilayers have aroused great interest due to their potential as a novel therapeutic strategy by disrupting cellular ion homeostasis.Based on the structural advantages of molecular rotaxanes,we herein show that two rotaxane-based transporters[2]R and[3]R induce coupled K^(+)/Cl^(-) channel transport by introducing Cl^(-) recognition sites in the thread and K^(+) binding group in the wheel,respectively.The welldesigned molecular structures allow the insertion of unimolecular rotaxanes into the lipid bilayer,thus achieving effective ion transport by means of thermodynamically controlled movement and driven by the difference in ion concentration inside and outside the vesicles.In addition,the use of a three-component rotaxane can accelerate ion transport through a cooperative shuttlerelay mechanism in which two wheels move along the thread in the lipid membrane,thereby enabling[3]R to have higher ion transport capacity.This work represents a major advance in the use of rotaxane molecules to accomplish more complex and effective tasks.
文摘Objective The present study investigated the sodium/iodide symporter mRNA expression in mouse lactating mammary gland cells under different iodine levels and the effects of thyroid-stimulating hormone(TSH),estradiol(E2)and prolactin(PRL)on NIS mRNA expression in mouse lactating mammary gland cells.
文摘Radioactive iodine(RAI)therapy remains a cornerstone in the management of differentiated thyroid cancer(DTC).The therapeutic efficacy of RAI depends on thyroid-stimulating hormone(TSH)-driven uptake via the sodium-iodide symporter(NIS).Although elevated TSH levels are routinely achieved before RAI administration,the circadian rhythm of TSH,which peaks overnight,is largely overlooked in treatment protocols.Aligning RAI administration with this physiological peak,particularly through evening dosing,may enhance iodine uptake,improve therapeutic efficacy,and reduce systemic exposure.Preclinical data and the known circadian regulation of TSH and NIS support this approach,although clinical validation is lacking.Evening dosing may also improve patient convenience and clinic workflow.However,logistical challenges and the absence of human trials present barriers to implementation.In this narrative review,we synthesize current evidence and tentatively propose the chronotherapeutic alignment of RAI timing as a novel,potentially impactful optimization strategy in DTC management.Prospective studies are needed to evaluate its clinical benefits and feasibility.
文摘The symbiotic association between reef-building corals and Symbiodiniaceae is pivotal for coral reef ecosystems,yet remains susceptible to environmental factors.Currently,there is a dearth of research examining seasonal fluctuations in coral-associated Symbiodiniaceae communities.In this study,we investigated the seasonal dynamics of Symbiodiniaceae communities associated with coral species in the Luhuitou coral reef using high-throughput sequencing techniques and SymPortal analytical framework.The results indicated that the genus Cladocopium exhibited dominance(averaging 82%),followed by Durusdinium(18%)and Breviolum(0.01%)within the examined coral species.Among the 521 Symbiodiniaceae ITS2 sequence types,C15 emerged as the prevalent type(13.24%),trailed by C3u(9.51%)and D1(8.57%).Interestingly,Symbiodiniaceae communities varied among different coral species.Pocillopora damicornis displayed a predominant association with Durusdinium,while Porites lutea,Goniastrea retiformis,Montipora truncata,Montipora aequituberculata,and Acropora divaricata were entirely dominated by the genus Cladocopium(100%),showcasing distinct host specificity.In the cases of Hydnophora exesa,Acropora latistella,Acropora digitifera,and seawater,both Cladocopium and Durusdinium were concurrently detected.Moreover,the diversity of Symbiodiniaceae associated with P.damicornis,P.lutea,G.retiformis,M.truncata,M.aequituberculata,and A.digitifera exhibited significant variations across different seasons.Notably,the results revealed that the alterations in Symbiodiniaceae community compositions were primarily driven by nutrient concentrations and seawater temperature.The network analysis of Symbiodiniaceae revealed the dominant Symbiodiniaceae types C15,C17f,C3u,C3,and D4 were exclusive.This study provided the seasonal variation characteristics of Symbiodiniaceae communities among different coral species,which may be a potential adaptive mechanism to environmental conditions.
