AIM:To develop a molecular therapy for pancreatic cancer, the insulin-like growth factor-I (IGF-I) signaling pathway was analyzed.METHODS: Pancreatic cancer cell lines (MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 a...AIM:To develop a molecular therapy for pancreatic cancer, the insulin-like growth factor-I (IGF-I) signaling pathway was analyzed.METHODS: Pancreatic cancer cell lines (MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 and KP-4) were cultured in media with 10 mL/L fetal bovine serum. Western blotting analysis was performed to clarify the expression of IGF-I receptor (IGF-IR). Picropodophyllin (PPP), a specific inhibitor of IGF-IR, LY294002, a specific inhibitor of phosphatidylinositol3 kinase (PI3K), and PD98059, a specific inhibitor of mitogen-activated protein kinase, were added to the media. After 72 h, a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay was performed to analyze cell proliferation. A wound assay was performed to analyze cell motility with hematoxylin and eosin (HE) staining 48 h after addition of each inhibitor. RESULTS: All cell lines clearly expressed not only IGF-IR but also phosphorylated IGF-IR. PPP significantly suppressed proliferation of MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 and KP-4 cells to 36.9% ± 2.4% (mean ± SD), 30.9% ± 5.5%, 23.8% ± 3.9%, 37.1% ± 5.3%, 10.4% ± 4.5%, 52.5% ± 4.5% and 22.6% ± 0.4%, at 2 μmol/L, respectively (P < 0.05). LY294002 significantly suppressed proliferation of MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 and KP-4 cells to 44.4% ± 7.6%, 32.9% ± 8.2%, 53.9% ± 8.0%, 52.8% ± 4.0%, 32.3% ± 4.2%, 51.8% ± 4.5%, and 30.6% ± 9.4%, at 50 μmol/L, respectively (P < 0.05). PD98059 did not significantly suppress cell proliferation. PPP at 2 μmol/L suppressed motility of MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 and KP-4 cells to 3.0% ± 0.2%, 0%, 0%, 2.0% ± 0.1%, 5.0% ± 0.2%, 3.0% ± 0.1%, and 5.0% ± 0.2%, respectively (P < 0.05). LY294002 at 50 μmol/L suppressed motility of MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 and KP-4 to 3.0% ± 0.2%, 0%, 3.0% ± 0.2%, 0%, 0%, 0% and 3% ± 0.1%, respectively (P < 0.05). PD980509 at 20 μmol/L did not suppress motility. Cells were observed by microscopy to analyze the morphological changes induced by the inhibitors. Cells in medium treated with 2 μmol/L PPP or 50 μmol/L LY294002 had pyknotic nuclei, whereas those in medium with 20 μmol/L PD98059 did not show apoptosis.CONCLUSION: IGF-IR and PI3K are good candidates for molecular therapy of pancreatic cancer.展开更多
Background Hyperinsulinemia, insulin-like growth factor （IGF）-I and -Ⅱ （IGF-Ⅱ） are associated with increased risk of endometrial carcinoma. Insulin receptor isoform A （IR-A） is more frequently expressed in end...Background Hyperinsulinemia, insulin-like growth factor （IGF）-I and -Ⅱ （IGF-Ⅱ） are associated with increased risk of endometrial carcinoma. Insulin receptor isoform A （IR-A） is more frequently expressed in endombtrial carcinoma than in normal endometrial tissues. To better understand their roles in endometrial carcinoma, we investigated the effects of insulin, IGF-I, and IGF-II in endometrial carcinomas cells with different IR-A expression levels. Methods To explore the role of IR-A in mediating the activity of IGF-I, IGF-II, and insulin, we investigate the cellular proliferation of endometrial carcinoma cell lines RL95-2 and RL95-2-1R-A by MTS assays. Then we examined the protein kinase Akt phosphorylation and extracellular signal-regulated kinase （ERK） 1/2 phosphorylation in both cell lines by Western blotting. The effect of IGF-II and AG1024 on cell cycle progression and apoptosis was assessed by fiowcytometry. To examine whether the effects of IGFs were mediated by IR-A, we blocked IGF-I receptor （IGF-IR） in both cell lines using AG1024, an IGF-IR-specific inhibitor. Results IGF-I and IGF-II significantly enhanced proliferation of both cell lines （P 〈0.05）. By contrast, insulin significantly increased proliferation of RL95-2-1R-A cells only （P 〈0.05）. IGF-I and IGF-II significantly increased pAkt levels in RL95-2 cells and pERK1/2 levels in RL95-2-1R-A cells （all, P 〈0.05）. Insulin increased pERK1/2 levels in RL95-2-1R-A cells only （P 〈0.05）. LY294002 and PD98059 inhibited the specific signaling activities and cellular proliferation. After AG1024 pretreatment, neither IGF-I nor IGF-II affected pAkt levels in RL95-2 cells. IGF-II, but not IGF-I, increased pERK1/2 levels in RL95-2-1R-A cells. After AG1024 pretreatment, the proliferation rate and DNA content corresponding to the S phase increased and apoptosis decreased significantly in IGF-II-treated RL95-2-1R-A cells only （P 〈0.05）. Conclusions The proliferation effect of insulin is mediated by IR-A. When IR-A dominates in a cell line, IGF-II activated cell proliferation mainly through the ERKI/2 pathway. On the other hand, IGF-II activated cell proliferation mainly through the Akt pathway. IR-A can at least partly mediate the proliferative and anti-apoptotic effects of IGF-II through the ERKI/2 pathway.展开更多
目的了解长三角地区耐多药/利福平耐药结核病(multidrug/rifampicin-resistant tuberculosis,MDR/RRTB)患者灾难性卫生支出(catastrophic health expenditure,CHE)的发生情况,识别发生CHE的高风险人群及风险时点。方法采用分层整群抽样...目的了解长三角地区耐多药/利福平耐药结核病(multidrug/rifampicin-resistant tuberculosis,MDR/RRTB)患者灾难性卫生支出(catastrophic health expenditure,CHE)的发生情况,识别发生CHE的高风险人群及风险时点。方法采用分层整群抽样的方法,选取长三角地区三省一市的10家耐药结核病定点医院为研究现场,研究对象为2017年7月—2019年6月在选定医院登记治疗的MDR/RR-TB患者。通过自行设计的调查问卷收集患者的基本人口学信息,导出医院医疗信息系统和收费系统中结核病诊疗相关信息及费用。以直接医疗总费用、报销费用及CHE发生率等描述MDR/RR-TB患者的疾病经济负担,并采用二元Logistic回归模型分析CHE的影响因素,逐月计算累计CHE发生率以确定风险点。结果共发放问卷387份,回收有效问卷344份,有效回收率88.89%。参与调查的患者中CHE发生率为62.79%,C地(OR=4.50,95%CI:1.08~18.76)和D地(OR=10.22,95%CI:1.70~61.39)的患者相较于A地更容易发生CHE。家庭年总收入高(OR=0.01,95%CI:0.01~0.05)、有医疗保险(OR=0.17,95%CI:0.04~0.62)是CHE发生的保护因素,而治疗期间住院(OR=8.06,95%CI:3.26~22.68)则是CHE发生的风险因素。调查对象接受抗结核治疗的前4个月CHE发生率上升最快,4个月末累计CHE发生率为60.47%,5~6个月上升到61.63%,6个月后上升速度逐渐变缓并趋于平稳。结论MDR/RR-TB患者仍承受沉重的疾病经济负担,低文化水平、家庭年总收入低、无医保及住院患者为CHE高风险人群。需加强MDR/RRTB患者治疗期间尤其是治疗前4个月的经济保护,针对不同患者优化医保与减免政策,以降低CHE发生率。展开更多
文摘AIM:To develop a molecular therapy for pancreatic cancer, the insulin-like growth factor-I (IGF-I) signaling pathway was analyzed.METHODS: Pancreatic cancer cell lines (MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 and KP-4) were cultured in media with 10 mL/L fetal bovine serum. Western blotting analysis was performed to clarify the expression of IGF-I receptor (IGF-IR). Picropodophyllin (PPP), a specific inhibitor of IGF-IR, LY294002, a specific inhibitor of phosphatidylinositol3 kinase (PI3K), and PD98059, a specific inhibitor of mitogen-activated protein kinase, were added to the media. After 72 h, a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay was performed to analyze cell proliferation. A wound assay was performed to analyze cell motility with hematoxylin and eosin (HE) staining 48 h after addition of each inhibitor. RESULTS: All cell lines clearly expressed not only IGF-IR but also phosphorylated IGF-IR. PPP significantly suppressed proliferation of MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 and KP-4 cells to 36.9% ± 2.4% (mean ± SD), 30.9% ± 5.5%, 23.8% ± 3.9%, 37.1% ± 5.3%, 10.4% ± 4.5%, 52.5% ± 4.5% and 22.6% ± 0.4%, at 2 μmol/L, respectively (P < 0.05). LY294002 significantly suppressed proliferation of MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 and KP-4 cells to 44.4% ± 7.6%, 32.9% ± 8.2%, 53.9% ± 8.0%, 52.8% ± 4.0%, 32.3% ± 4.2%, 51.8% ± 4.5%, and 30.6% ± 9.4%, at 50 μmol/L, respectively (P < 0.05). PD98059 did not significantly suppress cell proliferation. PPP at 2 μmol/L suppressed motility of MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 and KP-4 cells to 3.0% ± 0.2%, 0%, 0%, 2.0% ± 0.1%, 5.0% ± 0.2%, 3.0% ± 0.1%, and 5.0% ± 0.2%, respectively (P < 0.05). LY294002 at 50 μmol/L suppressed motility of MIA-Paca2, NOR-P1, PANC-1, PK-45H, PK-1, PK-59 and KP-4 to 3.0% ± 0.2%, 0%, 3.0% ± 0.2%, 0%, 0%, 0% and 3% ± 0.1%, respectively (P < 0.05). PD980509 at 20 μmol/L did not suppress motility. Cells were observed by microscopy to analyze the morphological changes induced by the inhibitors. Cells in medium treated with 2 μmol/L PPP or 50 μmol/L LY294002 had pyknotic nuclei, whereas those in medium with 20 μmol/L PD98059 did not show apoptosis.CONCLUSION: IGF-IR and PI3K are good candidates for molecular therapy of pancreatic cancer.
基金This work was supported by grants from the Specialized Research Fund for the Doctoral Program of Higher Education （No. 200800010095） and the National Natural Science Foundation of China （No. 30973181）.
文摘Background Hyperinsulinemia, insulin-like growth factor （IGF）-I and -Ⅱ （IGF-Ⅱ） are associated with increased risk of endometrial carcinoma. Insulin receptor isoform A （IR-A） is more frequently expressed in endombtrial carcinoma than in normal endometrial tissues. To better understand their roles in endometrial carcinoma, we investigated the effects of insulin, IGF-I, and IGF-II in endometrial carcinomas cells with different IR-A expression levels. Methods To explore the role of IR-A in mediating the activity of IGF-I, IGF-II, and insulin, we investigate the cellular proliferation of endometrial carcinoma cell lines RL95-2 and RL95-2-1R-A by MTS assays. Then we examined the protein kinase Akt phosphorylation and extracellular signal-regulated kinase （ERK） 1/2 phosphorylation in both cell lines by Western blotting. The effect of IGF-II and AG1024 on cell cycle progression and apoptosis was assessed by fiowcytometry. To examine whether the effects of IGFs were mediated by IR-A, we blocked IGF-I receptor （IGF-IR） in both cell lines using AG1024, an IGF-IR-specific inhibitor. Results IGF-I and IGF-II significantly enhanced proliferation of both cell lines （P 〈0.05）. By contrast, insulin significantly increased proliferation of RL95-2-1R-A cells only （P 〈0.05）. IGF-I and IGF-II significantly increased pAkt levels in RL95-2 cells and pERK1/2 levels in RL95-2-1R-A cells （all, P 〈0.05）. Insulin increased pERK1/2 levels in RL95-2-1R-A cells only （P 〈0.05）. LY294002 and PD98059 inhibited the specific signaling activities and cellular proliferation. After AG1024 pretreatment, neither IGF-I nor IGF-II affected pAkt levels in RL95-2 cells. IGF-II, but not IGF-I, increased pERK1/2 levels in RL95-2-1R-A cells. After AG1024 pretreatment, the proliferation rate and DNA content corresponding to the S phase increased and apoptosis decreased significantly in IGF-II-treated RL95-2-1R-A cells only （P 〈0.05）. Conclusions The proliferation effect of insulin is mediated by IR-A. When IR-A dominates in a cell line, IGF-II activated cell proliferation mainly through the ERKI/2 pathway. On the other hand, IGF-II activated cell proliferation mainly through the Akt pathway. IR-A can at least partly mediate the proliferative and anti-apoptotic effects of IGF-II through the ERKI/2 pathway.
文摘目的了解长三角地区耐多药/利福平耐药结核病(multidrug/rifampicin-resistant tuberculosis,MDR/RRTB)患者灾难性卫生支出(catastrophic health expenditure,CHE)的发生情况,识别发生CHE的高风险人群及风险时点。方法采用分层整群抽样的方法,选取长三角地区三省一市的10家耐药结核病定点医院为研究现场,研究对象为2017年7月—2019年6月在选定医院登记治疗的MDR/RR-TB患者。通过自行设计的调查问卷收集患者的基本人口学信息,导出医院医疗信息系统和收费系统中结核病诊疗相关信息及费用。以直接医疗总费用、报销费用及CHE发生率等描述MDR/RR-TB患者的疾病经济负担,并采用二元Logistic回归模型分析CHE的影响因素,逐月计算累计CHE发生率以确定风险点。结果共发放问卷387份,回收有效问卷344份,有效回收率88.89%。参与调查的患者中CHE发生率为62.79%,C地(OR=4.50,95%CI:1.08~18.76)和D地(OR=10.22,95%CI:1.70~61.39)的患者相较于A地更容易发生CHE。家庭年总收入高(OR=0.01,95%CI:0.01~0.05)、有医疗保险(OR=0.17,95%CI:0.04~0.62)是CHE发生的保护因素,而治疗期间住院(OR=8.06,95%CI:3.26~22.68)则是CHE发生的风险因素。调查对象接受抗结核治疗的前4个月CHE发生率上升最快,4个月末累计CHE发生率为60.47%,5~6个月上升到61.63%,6个月后上升速度逐渐变缓并趋于平稳。结论MDR/RR-TB患者仍承受沉重的疾病经济负担,低文化水平、家庭年总收入低、无医保及住院患者为CHE高风险人群。需加强MDR/RRTB患者治疗期间尤其是治疗前4个月的经济保护,针对不同患者优化医保与减免政策,以降低CHE发生率。
