BACKGROUND Hepatocellular carcinoma(HCC)is at the forefront of the global spectrum of cancer incidence and mortality,with conventional therapies like tyrosine kinase inhibitors limited by resistance.Recent studies hav...BACKGROUND Hepatocellular carcinoma(HCC)is at the forefront of the global spectrum of cancer incidence and mortality,with conventional therapies like tyrosine kinase inhibitors limited by resistance.Recent studies have highlighted the promising anticancer effects of nitidine chloride(NC)against HCC.SAC3 domain containing 1(SAC3D1)is critical for centrosome replication and spindle formation.However,research on SAC3D1 in HCC and NC remains limited.AIM To investigate the mechanisms underlying SAC3D1’s role in HCC progression and evaluated its potential as a therapeutic target of NC.METHODS RNA sequencing(RNA-seq)identified SAC3D1 expression changes in HCC cells after NC treatment.Molecular docking was further employed to validate the direct binding between NC and SAC3D1.Additionally,HCC multicenter data(The Cancer Genome Atlas_GTEx,ArrayExpress),pathway analysis,Pearson correlation analysis and SAC3D1 in vitro knockdown experiments were integrated to explore the molecular mechanisms underlying SAC3D1's involvement in HCC progression.RESULTS RNA-seq showed that NC treatment significantly downregulated SAC3D1 expression[log2(fold change)=-0.95,P<0.05],with molecular docking revealing that NC directly bound to SAC3D1 proteins(binding energy:-9.7 kcal/mol).Enrichment analysis showed that most pathways were closely related to the cell cycle.Pearson correlation analysis indicated that SAC3D1 and cell cycle genes were significantly positively correlated(correlation coefficient≥0.3,P<0.05).SAC3D1 knockdown inhibited HCC cell invasion,migration,and proliferation by arresting cells in the S and G2/M phases.Flow cytometry confirmed that after SAC3D1 knockdown,the early and total apoptosis percentage of HCC cells decreased,while the late apoptosis percentage increased.CONCLUSION As a potential target of NC,SAC3D1 may inhibit HCC progression through cell cycle regulation following its downregulation by NC.展开更多
BACKGROUND The prevalence of colorectal cancer(CRC)in younger people is increasing.Despite advances in precision medicine,the challenges of drug resistance and high costs persist.Nitidine chloride(NC)has pharmacologic...BACKGROUND The prevalence of colorectal cancer(CRC)in younger people is increasing.Despite advances in precision medicine,the challenges of drug resistance and high costs persist.Nitidine chloride(NC)has pharmacological potential,and kinesin family member 20A(KIF20A)is overexpressed in various tumors;however,their interaction in CRC remains unexplored.AIM To investigate the KIF20A expression characteristics in CRC cells and determine whether it is a potential target gene for NC in inhibiting CRC treatment.METHODS Single-cell RNA sequencing(scRNA-seq),spatial transcriptomics,and mRNA expression profiling were used to analyze KIF20A expression in CRC cells.Immunohistochemical staining was used to verify KIF20A expression in 416 clinical samples(208 CRC tissue samples and 208 noncancerous control tissue samples).Clustered regularly interspaced short palindromic repeats(CRISPR)technology was used to evaluate the impact of knocking out KIF20A on CRC cell growth.Molecular docking was applied to analyze NC–KIF20A binding.Finally,RNA sequencing and functional enrichment analysis were performed to explore the mechanism of action of NC in CRC cells.RESULTS Treating HCT116 cells with NC was found to significantly downregulate KIF20A(P<0.05),and the molecular docking analysis revealed high-affinity binding between NC and KIF20A(binding energy=-9.6 kcal/mol).The scRNA-seq,spatial transcriptomics,and mRNA expression profiling results confirmed the significantly high expression of KIF20A in CRC tissues(standardized mean difference=1.33,95%confidence interval:0.885-1.77,summary receiver operating characteristic curve area=0.94).The immunohistochemical analysis of the clinical samples showed high KIF20A expression in the CRC tissues(P<0.05),with significant correlation between the level of expression and gender,tumor size,and tumor grade(P<0.05).Knocking out KIF20A significantly inhibited the growth of various CRC cell lines(CRISPR score<-0.3).The functional enrichment analysis indicated that NC may inhibit CRC by disrupting several biological processes,such as mitotic nuclear division,chromosome segregation,and microtubule binding.CONCLUSION Our results indicate that NC binds to KIF20A with high affinity and downregulates its expression in CRC cells,leading to reduced proliferation.Hence,NC has promise as a therapeutic agent in the treatment of CRC,and targeting KIF20A also has potential as a therapeutic strategy.Further KIF20A knockout studies are needed to confirm the binding specificity and mechanistic roles of NC in CRC.展开更多
Evaluating toxicity and decoding the underlying mechanisms of active compounds are crucial for drug development.In this study,we present an innovative,integrated approach that combines air flowassisted desorption elec...Evaluating toxicity and decoding the underlying mechanisms of active compounds are crucial for drug development.In this study,we present an innovative,integrated approach that combines air flowassisted desorption electrospray ionization mass spectrometry imaging(AFADESI-MSI),time-of-flight secondary ion mass spectrometry(ToF-SIMS),and spatial metabolomics to comprehensively investigate the nephrotoxicity and underlying mechanisms of nitidine chloride(NC),a promising anti-tumor drug candidate.Our quantitive AFADESI-MSI analysis unveiled the region specific of accumulation of NC in the kidney,particularly within the inner cortex(IC)region,following single and repeated dose of NC.High spatial resolution ToF-SIMS analysis further allowed us to precisely map the localization of NC within the renal tubule.Employing spatial metabolomics based on AFADESI-MSI,we identified over 70 discriminating endogenous metabolites associated with chronic NC exposure.These findings suggest the renal tubule as the primary target of NC toxicity and implicate renal transporters(organic cation transporters,multidrug and toxin extrusion,and organic cation transporter 2(OCT2)),metabolic enzymes(protein arginine N-methyltransferase(PRMT)and nitric oxide synthase),mitochondria,oxidative stress,and inflammation in NC-induced nephrotoxicity.This study offers novel insights into NC-induced renal damage,representing a crucial step towards devising strategies to mitigate renal damage caused by this compound.展开更多
Toddalia asiatica Linn. is an important medicinal plant belonging to the family Rutaceae. The plant is well known for its antimalarial activity, which has been attributed to the presence of benzophenanthridine alkaloi...Toddalia asiatica Linn. is an important medicinal plant belonging to the family Rutaceae. The plant is well known for its antimalarial activity, which has been attributed to the presence of benzophenanthridine alkaloid nitidine in the roots of plants. A simple, rapid, sensitive, accurate, repeatable and robust HPTLC method has been developed and validated for the quantitative determination of nitidine in the dried roots and plant tissue culture extracts of T. asiatica. Nitidine was estimated at 332 nm by densitometry using Silica gel 60 F254 as stationary phase and chloroform:methanol (7:1, v/v), and as mobile phase. Linearity was observed in the concentration range of 25 -200 ng/spot for nitidine. The limit of detection and limit of quantitation were found to be 0.026 and 0.086 ng/spot respectively for nitidine. Developed method was validated according to the ICH guidelines with respect to precision, accuracy, specificity and robustness. The technique has been applied for the first time for the estimation of nitidine in roots and plant tissue culture extracts of T. asiatica. Statistical analysis data indicate the accuracy and reliability of the method.展开更多
Nitidine, Chelerythrine and Sanguinarine, all these three alkaloids are benzophenanthridine alkaloids. Nitidine was used as an anti-HIV, anti-malarial and anti-cancer. Chelerythrine had anti-cancer and anti-inflammato...Nitidine, Chelerythrine and Sanguinarine, all these three alkaloids are benzophenanthridine alkaloids. Nitidine was used as an anti-HIV, anti-malarial and anti-cancer. Chelerythrine had anti-cancer and anti-inflammatory activities. Sanguinarine was widely used as an anti-plaquestic and anti-cancer. High performance thin layer chromatography (HPTLC) method was used for simultaneous quantification of Nitidine, Chelerythrine and Sanguinarine in callus extract of Zanthoxylum rhetsa by using Silica gel 60 F254 as stationary phase and ethyl acetate:methanol:water:diethylamine (30:5:2:0.5 v/v) as mobile phase at 280 nm. The linearity concentration range was 5 - 160 μg/band of each alkaloid. The Rf values of Nitidine, Chelerythrine and Sanguinarine were found to be 0.28, 0.49 and 0.73. The limit of detection and limit of quantification were found to be 0.026, 0.088 μg/spot and 0.010 and 0.033 μg/spot, 0.0104 and 0.035 μg/spot respectively for Nitidine, Chelerythrine and Sanguinarine. HPTLC method was developed and validated according to ICH guidelines for simultaneous estimation of Nitidine, Chelerythrine and Sanguinarine and proved to be simple, specific, accurate, robust and rapid.展开更多
BACKGROUND Colorectal cancer(CRC)is the third most common cancer globally,causing over 900000 deaths annually.Risk factors include aging,diet,obesity,sedentary lifestyle,tobacco use,genetic predisposition,and inflamma...BACKGROUND Colorectal cancer(CRC)is the third most common cancer globally,causing over 900000 deaths annually.Risk factors include aging,diet,obesity,sedentary lifestyle,tobacco use,genetic predisposition,and inflammatory bowel disease.Despite current treatments,survival rates for advanced CRC remain low,highlighting the need for better therapeutic strategies.AIM To evaluate both the clinical significance and the pathological implications of the Kinesin family member 14(KIF14)expression within CRC specimens.Additionally,this study aims to investigate the interaction between nitidine chloride(NC)and KIF14,considering their potential as therapeutic targets.METHODS The expression of the KIF14 protein in CRC was analyzed using immunohistochemical staining.The integration of multicenter high-throughput data facilitated the calculation of the standardized mean difference(SMD)for KIF14 mRNA levels.The assessment of clinical and pathological impact was enhanced by analyzing combined receiver operating characteristic curves,along with measures of sensitivity,specificity,and likelihood ratios.Additionally,clustered regularly interspaced short palindromic repeats knockout screening for cell growth and single-cell sequencing were employed to validate the significance of KIF14 expression in CRC.Survival analysis established the prognostic value of KIF14 in CRC.The molecular mechanism of NC against CRC was elucidated through whole-genome sequencing and enrichment analysis,and molecular docking was utilized to explore the targeting affinity between NC and KIF14.RESULTS KIF14 was highly expressed in 208 CRC patients.Data from 17 platforms involving 2436 CRC samples and 1320 noncancerous colorectal tissue controls indicated that KIF14 expression was significantly higher in CRC samples,with an SMD of 1.92(95%CI:1.49-2.35).The area under the curve was 0.94(95%CI:0.92-0.96),with a sensitivity of 0.85(95%CI:0.78-0.90)and a specificity of 0.90(95%CI:0.85-0.93).The positive and negative likelihood ratios were 8.38(95%CI:5.39-13.02)and 0.17(95%CI:0.11-0.26),respectively.At the single-cell level,significant overexpression of KIF14 was observed in CRC cells(P<0.001),with 35 CRC cell lines dependent on KIF14 for growth.The K-M plots demonstrated that KIF14 possesses prognostic value in CRC patients within the GSE71187 and GSE103679 datasets(P<0.05).Binding energy calculations indicated that KIF14 is a potential target for NC(binding energy:10.3 kcal/mol).CONCLUSION KIF14 promotes the growth of CRC cells and acts as an oncogenic factor,potentially serving as a therapeutic target for NC in the treatment of CRC.展开更多
This editorial provides insights into the pivotal role of checkpoint kinase 1(CHEK1)as both a biomarker and therapeutic target in colorectal cancer(CRC),based on findings from a recent study by Pang et al.Using single...This editorial provides insights into the pivotal role of checkpoint kinase 1(CHEK1)as both a biomarker and therapeutic target in colorectal cancer(CRC),based on findings from a recent study by Pang et al.Using single-cell RNA sequencing and immunohistochemistry,the study demonstrates significant CHEK1 overexpression in CRC tissues and identifies nitidine chloride as a potent CHEK1 inhibitor that disrupts DNA damage repair pathways.These findings underscore the therapeutic potential of CHEK1 inhibition and highlight the need for further research to address gaps in CRC treatment.展开更多
BACKGROUND Although transmembrane protein 106C(TMEM106C)has been elucidated to be overexpressed in cancers,its underlying mechanisms have not yet been fully understood.AIM To investigate the expression levels and mole...BACKGROUND Although transmembrane protein 106C(TMEM106C)has been elucidated to be overexpressed in cancers,its underlying mechanisms have not yet been fully understood.AIM To investigate the expression levels and molecular mechanisms of TMEM106C across 34 different cancer types,including liver hepatocellular carcinoma(LIHC).METHODS We analyzed TMEM106C expression patterns in pan-cancers using microenvironment cell populations counter to evaluate its association with the tumor microenvironment.Gene set enrichment analysis was conducted to identify molecular pathways related to TMEM106C.Chromatin immunoprecipitation followed by sequencing(ChIP-seq)analysis was conducted to identify upstream transcriptional regulators of TMEM106C.In LIHC,we examined mRNA profiles,performed in-house quantitative polymerase chain reaction,immunohistochemistry,and constructed a co-expression gene network.Functional assays,including cell counting kit-8,cell cycle,apoptosis,migration,and invasion,were conducted.The effect of nitidine chloride(NC)on LIHC xenograft was evaluated through RNA sequencing and molecular docking.Finally,potential therapeutic agents targeting TMEM106C were predicted.RESULTS TMEM106C was significantly overexpressed in 27 different cancer types and presaged poor prognosis in four of these types,including LIHC.Across pan-cancers,TMEM106C was inversely correlated to the abundances of immune and stromal cells.Furthermore,TMEM106C was significantly linked to cell cycle and DNA replication pathways in pan-cancers.ChIP-seq analysis predicted CCCTC-binding factor as a pivotal transcriptional factor targeting the TMEM106C gene in pan-cancers.Integrated analysis showed that TMEM106C was upregulated in 4657 LIHC compared with 3652 normal liver tissue[combined standardized mean difference=1.31(1.09,1.52)].Inhouse LIHC samples verified the expression status of TMEM106C.Higher TMEM106C expression signified worse survival conditions in LIHC patients treated with sorafenib,a tyrosine kinase inhibitor(TKI).Co-expressed analysis revealed that TMEM106C were significantly enriched in the cell cycle pathway.Knockout experiments demonstrated that TMEM106C plays a crucial role in LIHC cell proliferation,migration,and invasion,with cell cycle arrest occurring at the DNA synthesis phase,and increased apoptosis.Notably,TMEM106C upregulation was attenuated by NC treatment.Finally,TMEM106C expression levels were significantly correlated with the drug sensitivity of anti-hepatocellular carcinoma agents,including JNJ-42756493,a TKI agent.CONCLUSION Overexpressed TMEM106C was predicted as an oncogene in pan-cancers,which may serve as a promising therapeutic target for various cancers,including LIHC.Targeting TMEM106C could potentially offer a novel direction in overcoming TKI resistance specifically in LIHC.Future research directions include in-depth experimental validation and exploration of TMEM106C’s role in other cancer types.展开更多
BACKGROUND The stearoyl-coenzyme A desaturase(SCD)gene influences colorectal cancer(CRC)pathogenesis,with its expression linked to tumor cell survival and resistance,necessitating further investigation into its role i...BACKGROUND The stearoyl-coenzyme A desaturase(SCD)gene influences colorectal cancer(CRC)pathogenesis,with its expression linked to tumor cell survival and resistance,necessitating further investigation into its role in CRC.AIM To explore the clinical and pathological significance of SCD expression in CRC tissues and to evaluate the affinity between nitidine chloride(NC)and SCD as a target.METHODS Multi-center high-throughput data related to CRC were integrated to calculate the standardized mean difference of SCD mRNA expression levels.Immunohistochemical staining results,Clustered Regularly Interspaced Short Palindromic Repeats knockout screening results of cell growth,and single-cell sequencing were employed to verify the significance of SCD expression in CRC.The clinical and pathological significance of SCD was assessed using pooled receiver operating characteristic curves,sensitivity,specificity,and likelihood ratios.The molecular mechanism of NC against CRC was clarified using the SwissTarget Prediction and functional enrichment,and molecular docking techniques were utilized to explore the targeting affinity between NC and SCD.RESULTS Data from 18 platforms,including 2482 CRC samples and 1334 non-cancerous colorectal tissue controls.SCD expression was significantly upregulated in CRC,with a standardized mean difference of 2.05[95%confidence interval(CI):1.69-2.41].The area under the pooled receiver operating characteristic curve was 0.95(95%CI:0.92-0.96),with a sensitivity of 0.86(95%CI:0.81-0.90)and a specificity of 0.90(95%CI:0.87-0.93).Positive and negative likelihood ratios were 9.02(95%CI:6.49-12.51)and 0.15(95%CI:0.10-0.22),respectively.High SCD protein expression was noted in 208 CRC patients,significantly associated with vascular invasion(P<0.001).At the singlecell level,SCD was significantly overexpressed in CRC cells(P<0.001).A total of 33 CRC cell lines depended on SCD for growth.The potential mechanism of NC against CRC might involve modulation of the cell cycle,positioning SCD as a potential target for NC.CONCLUSION SCD promotes CRC cell growth and thus acts as an oncogenic factor,making it a potential therapeutic target for NC in CRC treatment.展开更多
Objective To study the pharmacokinetics of nitidine chloride（NC） in rat plasma after intragastrical（i.g.） administration. Methods A liquid chromatography-electrospray ionization-mass/mass sprectrometry（LC-ESI-MS...Objective To study the pharmacokinetics of nitidine chloride（NC） in rat plasma after intragastrical（i.g.） administration. Methods A liquid chromatography-electrospray ionization-mass/mass sprectrometry（LC-ESI-MS/MS） was used and carbamazepine was used as an intermal standard（I.S.）. The rat plasma samples were deproteinized with acetonitrile and the resultant supernatant was assayed on an analytical Diamonsil ^（TM）ODS C_（18） column（2.1 mm × 150 mm） equipped with a C_（18） guard column（4 mm × 20 mm） with a mobile phase of acetonitrile–10 mM ammonium acetate buffer–formic acid（35： 65： 0.2, v/v/v） at the flow rate of 0.25 mL/min. The LC–MS was carried out on a triple-quadrupole mass spectrometry equipped with an ESI and positive selected-ion monitoring. Target ions were monitored at [M-Cl]~＋ m/z 348.2 for NC and [M ＋ H]~＋ m/z237.2 for I.S., respectively. Results The simple one step deproteinize and rapid analysis method were successfully used in pharmacokinetic study on NC after i.g. administration. The linear relationship was good over the range of 2.5 – 1000.0 ng/ml（r^2 = 0.999 2） in rat plasma. The lower limit of quantification and detection were 2.5 and 1.6 ng/ml, respectively. The extraction recovery was in the range of 86.54 – 98.60%. The intra-and inter-day precisions（relative standard deviation） were less than 6.00%, with accuracies deviation between 89.40 to 95.57%. A two-compartment pharmacokinetic open model was proposed and validated to explain the apparent biphasic disposition of NC in rat plasma after i.g. administration. Conclusion This study was successfully applied to a pharmacokinetic study of NC in rats plasma following i.g. administration and could be used for preclinical and clinical pharmacokinetic evaluation of NC.展开更多
Objective:Alveolar macrophages(AMs)are involved in the development and progression of a variety of lung diseases.It is of great significance to explore the pathogenesis of diseases and evaluate the efficacy of drugs.H...Objective:Alveolar macrophages(AMs)are involved in the development and progression of a variety of lung diseases.It is of great significance to explore the pathogenesis of diseases and evaluate the efficacy of drugs.However,there is no standard process for extracting primary AM.Nitidine chloride(NC)is an alkaloid extracted from Zanthoxylum nitidum(Roxb.)DC.,which has an antitumour pharmacological effect.However,there is no evidence that NC has a direct effect on colorectal cancer cell lung metastasis.The purpose of this study was to establish a standard for the extraction of primary AM from mice and to investigate the pharmacodynamics of NC in mice with lung metastases to colorectal cancer.Methods:The standard for the extraction of mouse primary AM by lavage was established.Western blot and polymerase chain reaction were used to detect the regulatory mechanism of NC in the treatment of lung metastasis in mice by macrophage phenotype and glycolysis level.Results:The results showed that sufficient quantity and quality of primary AM could be obtained by optimizing extraction steps,and AM obtained by this method could accurately reflect disease progression.At the same time,NC can effectively reduce colorectal cancer lung metastasis in mice.From the mechanism,NC can inhibit the expression of M2 macrophage markers and the levels of mRNA and proteins of the glycolysis-limiting enzyme.Conclusions:Our results show that primary AM that accurately reflects disease and assesses pharmacological effects can be obtained using our established criteria.The inhibitory effect of NC on colorectal cancer lung metastasis may be attributed to its regulation of macrophage phenotype and glycolysis.展开更多
基金Supported by National Natural Science Foundation of China,No.82160762 and No.82460783Guangxi Medical University“Four New”Project,No.SX202403+2 种基金Innovation Project of Guangxi Graduate Education,No.JGY2023068Guangxi Higher Education Undergraduate Teaching Reform Project,No.2022JGA146China Undergraduate Innovation and Entrepreneurship Training Program,No.202310598045.
文摘BACKGROUND Hepatocellular carcinoma(HCC)is at the forefront of the global spectrum of cancer incidence and mortality,with conventional therapies like tyrosine kinase inhibitors limited by resistance.Recent studies have highlighted the promising anticancer effects of nitidine chloride(NC)against HCC.SAC3 domain containing 1(SAC3D1)is critical for centrosome replication and spindle formation.However,research on SAC3D1 in HCC and NC remains limited.AIM To investigate the mechanisms underlying SAC3D1’s role in HCC progression and evaluated its potential as a therapeutic target of NC.METHODS RNA sequencing(RNA-seq)identified SAC3D1 expression changes in HCC cells after NC treatment.Molecular docking was further employed to validate the direct binding between NC and SAC3D1.Additionally,HCC multicenter data(The Cancer Genome Atlas_GTEx,ArrayExpress),pathway analysis,Pearson correlation analysis and SAC3D1 in vitro knockdown experiments were integrated to explore the molecular mechanisms underlying SAC3D1's involvement in HCC progression.RESULTS RNA-seq showed that NC treatment significantly downregulated SAC3D1 expression[log2(fold change)=-0.95,P<0.05],with molecular docking revealing that NC directly bound to SAC3D1 proteins(binding energy:-9.7 kcal/mol).Enrichment analysis showed that most pathways were closely related to the cell cycle.Pearson correlation analysis indicated that SAC3D1 and cell cycle genes were significantly positively correlated(correlation coefficient≥0.3,P<0.05).SAC3D1 knockdown inhibited HCC cell invasion,migration,and proliferation by arresting cells in the S and G2/M phases.Flow cytometry confirmed that after SAC3D1 knockdown,the early and total apoptosis percentage of HCC cells decreased,while the late apoptosis percentage increased.CONCLUSION As a potential target of NC,SAC3D1 may inhibit HCC progression through cell cycle regulation following its downregulation by NC.
基金Supported by the Promoting Project of Basic Capacity for Young and Middle-aged University Teachers in Guangxi,No.2025KY0164Youth Science Foundation of Guangxi Medical University,No.GXMUYSF202423Guangxi Zhuang Autonomous Region Health Commission Scientific Research Project,No.Z-A20220415 and No.Z20210442.
文摘BACKGROUND The prevalence of colorectal cancer(CRC)in younger people is increasing.Despite advances in precision medicine,the challenges of drug resistance and high costs persist.Nitidine chloride(NC)has pharmacological potential,and kinesin family member 20A(KIF20A)is overexpressed in various tumors;however,their interaction in CRC remains unexplored.AIM To investigate the KIF20A expression characteristics in CRC cells and determine whether it is a potential target gene for NC in inhibiting CRC treatment.METHODS Single-cell RNA sequencing(scRNA-seq),spatial transcriptomics,and mRNA expression profiling were used to analyze KIF20A expression in CRC cells.Immunohistochemical staining was used to verify KIF20A expression in 416 clinical samples(208 CRC tissue samples and 208 noncancerous control tissue samples).Clustered regularly interspaced short palindromic repeats(CRISPR)technology was used to evaluate the impact of knocking out KIF20A on CRC cell growth.Molecular docking was applied to analyze NC–KIF20A binding.Finally,RNA sequencing and functional enrichment analysis were performed to explore the mechanism of action of NC in CRC cells.RESULTS Treating HCT116 cells with NC was found to significantly downregulate KIF20A(P<0.05),and the molecular docking analysis revealed high-affinity binding between NC and KIF20A(binding energy=-9.6 kcal/mol).The scRNA-seq,spatial transcriptomics,and mRNA expression profiling results confirmed the significantly high expression of KIF20A in CRC tissues(standardized mean difference=1.33,95%confidence interval:0.885-1.77,summary receiver operating characteristic curve area=0.94).The immunohistochemical analysis of the clinical samples showed high KIF20A expression in the CRC tissues(P<0.05),with significant correlation between the level of expression and gender,tumor size,and tumor grade(P<0.05).Knocking out KIF20A significantly inhibited the growth of various CRC cell lines(CRISPR score<-0.3).The functional enrichment analysis indicated that NC may inhibit CRC by disrupting several biological processes,such as mitotic nuclear division,chromosome segregation,and microtubule binding.CONCLUSION Our results indicate that NC binds to KIF20A with high affinity and downregulates its expression in CRC cells,leading to reduced proliferation.Hence,NC has promise as a therapeutic agent in the treatment of CRC,and targeting KIF20A also has potential as a therapeutic strategy.Further KIF20A knockout studies are needed to confirm the binding specificity and mechanistic roles of NC in CRC.
基金supported by the National Natural Science Foundation of China(Grant No.:21927808)the National Key Research and Development Program of China(Grant No.:2017YFC1704006).
文摘Evaluating toxicity and decoding the underlying mechanisms of active compounds are crucial for drug development.In this study,we present an innovative,integrated approach that combines air flowassisted desorption electrospray ionization mass spectrometry imaging(AFADESI-MSI),time-of-flight secondary ion mass spectrometry(ToF-SIMS),and spatial metabolomics to comprehensively investigate the nephrotoxicity and underlying mechanisms of nitidine chloride(NC),a promising anti-tumor drug candidate.Our quantitive AFADESI-MSI analysis unveiled the region specific of accumulation of NC in the kidney,particularly within the inner cortex(IC)region,following single and repeated dose of NC.High spatial resolution ToF-SIMS analysis further allowed us to precisely map the localization of NC within the renal tubule.Employing spatial metabolomics based on AFADESI-MSI,we identified over 70 discriminating endogenous metabolites associated with chronic NC exposure.These findings suggest the renal tubule as the primary target of NC toxicity and implicate renal transporters(organic cation transporters,multidrug and toxin extrusion,and organic cation transporter 2(OCT2)),metabolic enzymes(protein arginine N-methyltransferase(PRMT)and nitric oxide synthase),mitochondria,oxidative stress,and inflammation in NC-induced nephrotoxicity.This study offers novel insights into NC-induced renal damage,representing a crucial step towards devising strategies to mitigate renal damage caused by this compound.
文摘Toddalia asiatica Linn. is an important medicinal plant belonging to the family Rutaceae. The plant is well known for its antimalarial activity, which has been attributed to the presence of benzophenanthridine alkaloid nitidine in the roots of plants. A simple, rapid, sensitive, accurate, repeatable and robust HPTLC method has been developed and validated for the quantitative determination of nitidine in the dried roots and plant tissue culture extracts of T. asiatica. Nitidine was estimated at 332 nm by densitometry using Silica gel 60 F254 as stationary phase and chloroform:methanol (7:1, v/v), and as mobile phase. Linearity was observed in the concentration range of 25 -200 ng/spot for nitidine. The limit of detection and limit of quantitation were found to be 0.026 and 0.086 ng/spot respectively for nitidine. Developed method was validated according to the ICH guidelines with respect to precision, accuracy, specificity and robustness. The technique has been applied for the first time for the estimation of nitidine in roots and plant tissue culture extracts of T. asiatica. Statistical analysis data indicate the accuracy and reliability of the method.
文摘Nitidine, Chelerythrine and Sanguinarine, all these three alkaloids are benzophenanthridine alkaloids. Nitidine was used as an anti-HIV, anti-malarial and anti-cancer. Chelerythrine had anti-cancer and anti-inflammatory activities. Sanguinarine was widely used as an anti-plaquestic and anti-cancer. High performance thin layer chromatography (HPTLC) method was used for simultaneous quantification of Nitidine, Chelerythrine and Sanguinarine in callus extract of Zanthoxylum rhetsa by using Silica gel 60 F254 as stationary phase and ethyl acetate:methanol:water:diethylamine (30:5:2:0.5 v/v) as mobile phase at 280 nm. The linearity concentration range was 5 - 160 μg/band of each alkaloid. The Rf values of Nitidine, Chelerythrine and Sanguinarine were found to be 0.28, 0.49 and 0.73. The limit of detection and limit of quantification were found to be 0.026, 0.088 μg/spot and 0.010 and 0.033 μg/spot, 0.0104 and 0.035 μg/spot respectively for Nitidine, Chelerythrine and Sanguinarine. HPTLC method was developed and validated according to ICH guidelines for simultaneous estimation of Nitidine, Chelerythrine and Sanguinarine and proved to be simple, specific, accurate, robust and rapid.
基金Natural Science Foundation of Shandong Province,No.ZR2020QH185Scientific Research Nurturing Fund of The First Affiliated Hospital of Shandong First Medical University&Shandong Provincial Qianfoshan Hospital,No.QYPY2020NSFC0803+2 种基金Guangxi Zhuang Autonomous Region Health Commission Scientific Research Project,No.Z-A20220415Guangxi Medical University Teacher Teaching Ability Development Project,No.2022JFA02Guangxi Medical University Undergraduate Education and Teaching Reform Project,No.2023Y05.
文摘BACKGROUND Colorectal cancer(CRC)is the third most common cancer globally,causing over 900000 deaths annually.Risk factors include aging,diet,obesity,sedentary lifestyle,tobacco use,genetic predisposition,and inflammatory bowel disease.Despite current treatments,survival rates for advanced CRC remain low,highlighting the need for better therapeutic strategies.AIM To evaluate both the clinical significance and the pathological implications of the Kinesin family member 14(KIF14)expression within CRC specimens.Additionally,this study aims to investigate the interaction between nitidine chloride(NC)and KIF14,considering their potential as therapeutic targets.METHODS The expression of the KIF14 protein in CRC was analyzed using immunohistochemical staining.The integration of multicenter high-throughput data facilitated the calculation of the standardized mean difference(SMD)for KIF14 mRNA levels.The assessment of clinical and pathological impact was enhanced by analyzing combined receiver operating characteristic curves,along with measures of sensitivity,specificity,and likelihood ratios.Additionally,clustered regularly interspaced short palindromic repeats knockout screening for cell growth and single-cell sequencing were employed to validate the significance of KIF14 expression in CRC.Survival analysis established the prognostic value of KIF14 in CRC.The molecular mechanism of NC against CRC was elucidated through whole-genome sequencing and enrichment analysis,and molecular docking was utilized to explore the targeting affinity between NC and KIF14.RESULTS KIF14 was highly expressed in 208 CRC patients.Data from 17 platforms involving 2436 CRC samples and 1320 noncancerous colorectal tissue controls indicated that KIF14 expression was significantly higher in CRC samples,with an SMD of 1.92(95%CI:1.49-2.35).The area under the curve was 0.94(95%CI:0.92-0.96),with a sensitivity of 0.85(95%CI:0.78-0.90)and a specificity of 0.90(95%CI:0.85-0.93).The positive and negative likelihood ratios were 8.38(95%CI:5.39-13.02)and 0.17(95%CI:0.11-0.26),respectively.At the single-cell level,significant overexpression of KIF14 was observed in CRC cells(P<0.001),with 35 CRC cell lines dependent on KIF14 for growth.The K-M plots demonstrated that KIF14 possesses prognostic value in CRC patients within the GSE71187 and GSE103679 datasets(P<0.05).Binding energy calculations indicated that KIF14 is a potential target for NC(binding energy:10.3 kcal/mol).CONCLUSION KIF14 promotes the growth of CRC cells and acts as an oncogenic factor,potentially serving as a therapeutic target for NC in the treatment of CRC.
文摘This editorial provides insights into the pivotal role of checkpoint kinase 1(CHEK1)as both a biomarker and therapeutic target in colorectal cancer(CRC),based on findings from a recent study by Pang et al.Using single-cell RNA sequencing and immunohistochemistry,the study demonstrates significant CHEK1 overexpression in CRC tissues and identifies nitidine chloride as a potent CHEK1 inhibitor that disrupts DNA damage repair pathways.These findings underscore the therapeutic potential of CHEK1 inhibition and highlight the need for further research to address gaps in CRC treatment.
基金Supported by the National Natural Science Foundation of China,No.NSFC82160762,No.NSFC82460783Natural Science Foundation of Guangxi,No.2022GXNSFBA035657Innovation Project of Guangxi Graduate Education,No.JGY2023068,No.YCSW2023220.
文摘BACKGROUND Although transmembrane protein 106C(TMEM106C)has been elucidated to be overexpressed in cancers,its underlying mechanisms have not yet been fully understood.AIM To investigate the expression levels and molecular mechanisms of TMEM106C across 34 different cancer types,including liver hepatocellular carcinoma(LIHC).METHODS We analyzed TMEM106C expression patterns in pan-cancers using microenvironment cell populations counter to evaluate its association with the tumor microenvironment.Gene set enrichment analysis was conducted to identify molecular pathways related to TMEM106C.Chromatin immunoprecipitation followed by sequencing(ChIP-seq)analysis was conducted to identify upstream transcriptional regulators of TMEM106C.In LIHC,we examined mRNA profiles,performed in-house quantitative polymerase chain reaction,immunohistochemistry,and constructed a co-expression gene network.Functional assays,including cell counting kit-8,cell cycle,apoptosis,migration,and invasion,were conducted.The effect of nitidine chloride(NC)on LIHC xenograft was evaluated through RNA sequencing and molecular docking.Finally,potential therapeutic agents targeting TMEM106C were predicted.RESULTS TMEM106C was significantly overexpressed in 27 different cancer types and presaged poor prognosis in four of these types,including LIHC.Across pan-cancers,TMEM106C was inversely correlated to the abundances of immune and stromal cells.Furthermore,TMEM106C was significantly linked to cell cycle and DNA replication pathways in pan-cancers.ChIP-seq analysis predicted CCCTC-binding factor as a pivotal transcriptional factor targeting the TMEM106C gene in pan-cancers.Integrated analysis showed that TMEM106C was upregulated in 4657 LIHC compared with 3652 normal liver tissue[combined standardized mean difference=1.31(1.09,1.52)].Inhouse LIHC samples verified the expression status of TMEM106C.Higher TMEM106C expression signified worse survival conditions in LIHC patients treated with sorafenib,a tyrosine kinase inhibitor(TKI).Co-expressed analysis revealed that TMEM106C were significantly enriched in the cell cycle pathway.Knockout experiments demonstrated that TMEM106C plays a crucial role in LIHC cell proliferation,migration,and invasion,with cell cycle arrest occurring at the DNA synthesis phase,and increased apoptosis.Notably,TMEM106C upregulation was attenuated by NC treatment.Finally,TMEM106C expression levels were significantly correlated with the drug sensitivity of anti-hepatocellular carcinoma agents,including JNJ-42756493,a TKI agent.CONCLUSION Overexpressed TMEM106C was predicted as an oncogene in pan-cancers,which may serve as a promising therapeutic target for various cancers,including LIHC.Targeting TMEM106C could potentially offer a novel direction in overcoming TKI resistance specifically in LIHC.Future research directions include in-depth experimental validation and exploration of TMEM106C’s role in other cancer types.
基金Supported by Natural Science Foundation of Shandong Province,No.ZR2020QH185Scientific Research Nurturing Fund of the First Affiliated Hospital of Shandong First Medical University&Shandong Provincial Qianfoshan Hospital,No.QYPY2020NSFC0803Guangxi Zhuang Autonomous Region Health Commission Scientific Research Project,No.Z20210442.
文摘BACKGROUND The stearoyl-coenzyme A desaturase(SCD)gene influences colorectal cancer(CRC)pathogenesis,with its expression linked to tumor cell survival and resistance,necessitating further investigation into its role in CRC.AIM To explore the clinical and pathological significance of SCD expression in CRC tissues and to evaluate the affinity between nitidine chloride(NC)and SCD as a target.METHODS Multi-center high-throughput data related to CRC were integrated to calculate the standardized mean difference of SCD mRNA expression levels.Immunohistochemical staining results,Clustered Regularly Interspaced Short Palindromic Repeats knockout screening results of cell growth,and single-cell sequencing were employed to verify the significance of SCD expression in CRC.The clinical and pathological significance of SCD was assessed using pooled receiver operating characteristic curves,sensitivity,specificity,and likelihood ratios.The molecular mechanism of NC against CRC was clarified using the SwissTarget Prediction and functional enrichment,and molecular docking techniques were utilized to explore the targeting affinity between NC and SCD.RESULTS Data from 18 platforms,including 2482 CRC samples and 1334 non-cancerous colorectal tissue controls.SCD expression was significantly upregulated in CRC,with a standardized mean difference of 2.05[95%confidence interval(CI):1.69-2.41].The area under the pooled receiver operating characteristic curve was 0.95(95%CI:0.92-0.96),with a sensitivity of 0.86(95%CI:0.81-0.90)and a specificity of 0.90(95%CI:0.87-0.93).Positive and negative likelihood ratios were 9.02(95%CI:6.49-12.51)and 0.15(95%CI:0.10-0.22),respectively.High SCD protein expression was noted in 208 CRC patients,significantly associated with vascular invasion(P<0.001).At the singlecell level,SCD was significantly overexpressed in CRC cells(P<0.001).A total of 33 CRC cell lines depended on SCD for growth.The potential mechanism of NC against CRC might involve modulation of the cell cycle,positioning SCD as a potential target for NC.CONCLUSION SCD promotes CRC cell growth and thus acts as an oncogenic factor,making it a potential therapeutic target for NC in CRC treatment.
基金Natural Science Foundation of Guangxi Province of China(2010GXNSFA013187)Chinese Medicine Science and Technology Development of Guangxi Administration of TCM Project(GZKZ1143)+1 种基金Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources(Guangxi Normal University)Ministry of Education of China(CMEMR2012-B01)
文摘Objective To study the pharmacokinetics of nitidine chloride（NC） in rat plasma after intragastrical（i.g.） administration. Methods A liquid chromatography-electrospray ionization-mass/mass sprectrometry（LC-ESI-MS/MS） was used and carbamazepine was used as an intermal standard（I.S.）. The rat plasma samples were deproteinized with acetonitrile and the resultant supernatant was assayed on an analytical Diamonsil ^（TM）ODS C_（18） column（2.1 mm × 150 mm） equipped with a C_（18） guard column（4 mm × 20 mm） with a mobile phase of acetonitrile–10 mM ammonium acetate buffer–formic acid（35： 65： 0.2, v/v/v） at the flow rate of 0.25 mL/min. The LC–MS was carried out on a triple-quadrupole mass spectrometry equipped with an ESI and positive selected-ion monitoring. Target ions were monitored at [M-Cl]~＋ m/z 348.2 for NC and [M ＋ H]~＋ m/z237.2 for I.S., respectively. Results The simple one step deproteinize and rapid analysis method were successfully used in pharmacokinetic study on NC after i.g. administration. The linear relationship was good over the range of 2.5 – 1000.0 ng/ml（r^2 = 0.999 2） in rat plasma. The lower limit of quantification and detection were 2.5 and 1.6 ng/ml, respectively. The extraction recovery was in the range of 86.54 – 98.60%. The intra-and inter-day precisions（relative standard deviation） were less than 6.00%, with accuracies deviation between 89.40 to 95.57%. A two-compartment pharmacokinetic open model was proposed and validated to explain the apparent biphasic disposition of NC in rat plasma after i.g. administration. Conclusion This study was successfully applied to a pharmacokinetic study of NC in rats plasma following i.g. administration and could be used for preclinical and clinical pharmacokinetic evaluation of NC.
基金China Postdoctoral Science Foundation(No.2023M732627 and No.2024T170657)Natural Science Foundation of Tianjin Municipality(No:23JCZXJC00150 and 23JCJQJC00040)supported this work。
文摘Objective:Alveolar macrophages(AMs)are involved in the development and progression of a variety of lung diseases.It is of great significance to explore the pathogenesis of diseases and evaluate the efficacy of drugs.However,there is no standard process for extracting primary AM.Nitidine chloride(NC)is an alkaloid extracted from Zanthoxylum nitidum(Roxb.)DC.,which has an antitumour pharmacological effect.However,there is no evidence that NC has a direct effect on colorectal cancer cell lung metastasis.The purpose of this study was to establish a standard for the extraction of primary AM from mice and to investigate the pharmacodynamics of NC in mice with lung metastases to colorectal cancer.Methods:The standard for the extraction of mouse primary AM by lavage was established.Western blot and polymerase chain reaction were used to detect the regulatory mechanism of NC in the treatment of lung metastasis in mice by macrophage phenotype and glycolysis level.Results:The results showed that sufficient quantity and quality of primary AM could be obtained by optimizing extraction steps,and AM obtained by this method could accurately reflect disease progression.At the same time,NC can effectively reduce colorectal cancer lung metastasis in mice.From the mechanism,NC can inhibit the expression of M2 macrophage markers and the levels of mRNA and proteins of the glycolysis-limiting enzyme.Conclusions:Our results show that primary AM that accurately reflects disease and assesses pharmacological effects can be obtained using our established criteria.The inhibitory effect of NC on colorectal cancer lung metastasis may be attributed to its regulation of macrophage phenotype and glycolysis.
