沉默GTSE1通过Hippo-YAP/TAZ通路调控上皮间质转化对卵巢癌卡铂耐药的影响OA
Silencing GTSE1 regulates epithelial-mesenchymal transition via the Hippo-YAP/TAZ pathway to influence carboplatin resistance in ovarian cancer
目的 探究 G2/S 期应答相关蛋白 1(GTSE1)对卵巢癌卡铂耐药的作用及其机制.方法 (1)Western blot检测人卵巢癌细胞株OVCAR3、OV3R-CBP 和COC1 中GTSE1 表达量.(2)取OV3R-CBP细胞,转染si-NC或si-GTSE1,RT-qPCR及Western blot检测细胞中GTSE1 mRNA及蛋白表达量.分别给予 0、10、20、40、80 和 160 μmol/L卡铂处理,比较si-NC和si-GTSE1 的半数抑制浓度(IC50)和耐药指数.(3)分别给予 OV3R-CBP 细胞 70 μmol/L 卡铂(CBP 组)、70 μmol/L 卡铂+10 μmol/L LY2109761(CBP+LY2109761 组)、70 μmol/L卡铂+转染si-NC(CBP+si-NC组)、70 μmol/L卡铂+转染si-GTSE1(CBP+si-GTSE1组)、70 μmol/L卡铂+转染si-GTSE1+1 μmol/L XMU-MP-1(CBP+si-GTSE1+XMU-MP-1组)处理.Western blot检测E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、Yes相关蛋白(YAP)和转录共激活因子PDZ结合基序(TAZ)蛋白表达量.CCK-8及流式细胞术检测细胞增殖抑制及凋亡情况.结果 (1)人卵巢癌卡铂耐药细胞株OV3R-CBP组中GTSE1 表达量高于OVCAR3 组和COC1 组(P<0.05).(2)si-GTSE1 组的GTSE1 mRNA及蛋白表达量均低于si-NC组(P<0.05).si-GTSE1 组的卡铂的IC50 值和耐药指数均低于si-NC组(P<0.05).(3)与CBP+si-NC组相比,CBP+si-GTSE1 组的YAP、TAZ和Vimentin表达量均降低,细胞增殖抑制率、凋亡率、pYAP/YAP和pTAZ/TAZ值及E-cadherin表达量均升高(P<0.05).与CBP+si-GTSE1 组相比,CBP+si-GTSE1+XMU-MP-1组的YAP、TAZ和Vimentin表达量均升高,细胞增殖抑制率、凋亡率、pYAP/YAP和pTAZ/TAZ值及E-cadherin表达量均降低(P<0.05).结论 沉默GTSE1 可通过激活Hippo-YAP/TAZ通路抑制上皮间质转化,逆转OV3R-CBP细胞对卡铂的耐药性.
Objective To elucidate the role of G2/S phase expressed 1(GTSE1)in carboplatin resistance mechanisms in ovarian cancer.Methods(1)GTSE1 protein expression was compared between carboplatin-sensitive(OVCAR3,OV3R-CBP,COC1)cell lines by Western blot.(2)GTSE1 was then silenced in OV3R-CBP cells using small interfering(si)RNA transfection si-GTSE1 or si-normal control(NC),and the knockdown efficiency was confirmed by quantitative reverse transcription-polymerase chain reaction and Western blot.Carboplatin dose-response assays(0,10,20,40,80 and 160 μmol/L)were conducted to determine the half-maximal inhibitory concentration(IC50)and drug resistance index.(3)OV3R-CBP cells were treated with various combinations:carboplatin alone(70 μmol/L,CBP group),carboplatin plus LY2109761(10 μmol/L,CBP+si+LY2109261 group),carboplatin plus si-NC(CBP+si-NC group),carboplatin plus si-GTSE1(CBP+si-GTSE1 group),and a triple combination of carboplatin,si-GTSE1 and XMU-MP-1(1 μmol/L,CBP+si-GTSE1+XMU-MP-1 group).Expression levels of E-cadherin,Vimentin,Yes-associated protein(YAP)and transcriptional co-activator with PDZ-binding motif(TAZ)were detected by Western blot.Cell proliferation inhibition and apoptosis were assessed by Cell Counting Kit-8 assay and flow cytometry.Results(1)GTSE1 expression was significantly elevated in OV3R-CBP relative to OVCAR3/COC1 cells(P<0.05).(2)Knockdown of GTSE1 significantly decreased GTSE1 mRNA/protein levels and carboplatin IC50 and resistance index(P<0.05).(3)Silencing GTSE1 in the presence of carboplatin suppressed YAP,TAZ,and Vimentin expression,and enhanced E-cadherin expression,YAP/TAZ phosphorylation,cell proliferation inhibition,and apoptosis(P<0.05).Hippo pathway inhibition using XMU-MP-1 reversed these effects,indicating pathway-specific modulation.Conclusions Silencing GTSE1 reverses carboplatin resistance in ovarian cancer cells by activating the Hippo-YAP/TAZ pathway and inhibiting epithelial-mesenchymal transition.Targeting GTSE1 restores chemosensitivity via reactivation of the Hippo pathway,offering a potential therapeutic strategy for overcoming platinum resistance in ovarian cancer.
李娅茹;孙丽妍;宋永祯;林萍;陈欢
衡水市人民医院妇科,河北 衡水 053000衡水市人民医院妇科,河北 衡水 053000衡水市人民医院妇科,河北 衡水 053000衡水市人民医院妇科,河北 衡水 053000衡水市人民医院妇科,河北 衡水 053000
医药卫生
卵巢癌GTSE1YAPTAZ上皮间质转化卡铂耐药
ovarian cancerGTSE1YAPTAZepithelial-mesenchymal transitioncarboplatinresistance
《中国比较医学杂志》 2026 (1)
34-42,9
河北省医学科学研究课题计划项目(20211492).
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