奥美拉唑抑制miR-214-3p介导自噬提高上皮性卵巢癌细胞对顺铂的敏感性  

Omeprazole Enhances Cisplatin Sensitivity Through Inhibition of miR-214-3p Mediated Autophagy in Epithelial Ovarian Cancer Cells

作　　者：蔡文 季维雪 肖兰 江飞云 陈文刚 陶云松 Cai Wen;Ji Weixue;Xiao Lan(Department of Gynecology,Wuhu Hospital,East China Normal University(The Second People’s Hospital of Wuhu),Wuhu 241001,China;Department of Gynecology and Obstetrics,the First Affiliated Hospital of University of Science and Technology of China,Hefei 230022,China)

机构地区：[1]华东师范大学附属芜湖医院(芜湖市第二人民医院)妇科,芜湖241001 [2]中国科学技术大学附属第一医院妇产科,合肥230022 [3]皖南医学院第二附属医院药剂科,芜湖241001 [4]华东师范大学附属芜湖医院(芜湖市第二人民医院)药剂科,芜湖241001

出　　处：《华中科技大学学报（医学版）》2024年第1期90-93,109,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：安徽省卫生健康委员会科研项目(No.AHWJ2022a028)。

摘　　要：目的 探讨奥美拉唑(omeprazole, OME)能否通过抑制自噬提高人卵巢上皮性癌(卵巢癌,EOC)细胞对顺铂的敏感性及其可能机制。方法 原位杂交及免疫组化检测卵巢癌组织中miR-214-3p及自噬标记物p62表达;Pearson相关分析探讨R-214-3p和p62在EOC组织中表达量的相关性;CCK-8法检测各组细胞顺铂半数抑制浓度(IC50);实时定量PCR(qRT-PCR)分析miR-214-3p及多药耐药基因-1(MDR1)表达;蛋白质印迹法检测p62及耐药蛋白p-gp表达。结果 在43例卵巢癌患者的样本中,23例(53.5%)miR-214-3p表达阳性,26例(60.5%)p62表达阳性,miR-214-3p在铂相对耐药EOC中表达低于铂敏感EOC(P<0.05);相反,p62在铂相对耐药EOC中表达明显高于铂敏感EOC(P<0.01)。miR-214-3p和p62在卵巢癌中表达呈负相关(r=-0.387,P=0.005);OME(150μmol/L)预处理后,OV2008及C13K细胞对顺铂IC50均有不同程度降低,尤以顺铂耐药株C13K更为显著(P<0.01)。与空白对照组C13K及OV2008细胞比较,顺铂作用后,C13K及OV2008细胞中miR-214-3p相对表达下降,MDR1基因在mRNA及蛋白水平表达均升高,p62蛋白表达升高(均P<0.01);相反,OME(150μmol/L)预处理可使C13K及OV2008细胞对顺铂敏感性增加,细胞中miR-214-3p相对表达明显升高、MDR1在蛋白及mRNA水平表达降低、p62蛋白表达下降(均P<0.05)。结论 OME预处理可提高卵巢癌细胞对顺铂敏感性,其机制与抑制miR-214-3p介导的自噬有关。Objective To investigate whether omeprazole(OME)can enhance the sensitivity of epithelial ovarian cancer(EOC)cells to cisplatin(DDP)by inhibition of autophagy and to elucidate its possible mechanism.Methods Color in situ hybridization(CISH)and immunohistochemistry were applied to detect the expression of miR-214-3p and autophagy specific markers p62in EOC tissues,respectively.Pearson analysis showed the correlation between miR-214-3p and p62expression levels in EOC.The half concentration(IC50)of DDP was determined by CCK-8method.The mRNA expressions of miR-214-3p and multidrug resistance gene 1(MDR1),the protein levels of p-gp and p62were measured by using real-time quantitative PCR(qRTPCR)and Western blot,respectively.Results In 43cases,the expressions of miR-214-3p and p62were 53.5%(23/43)and 60.5%(26/43)in patients with ovarian carcinoma,respectively.miR-214-3p was downregulated in platinum-relatively resistant OC tissue(P<0.05).On the contrary,p62was upregulated in platinum-relatively resistant OC tissue(P<0.01).In ovarian cancer,the negative expression of miR-214-3p was closely related with p62(r=0.238,P<0.05).After OME(150μmol/L)pretreatment,varying degrees of decrease was observed in cisplatin IC50OV2008and C13Kcells,especially cisplatin resistant strain C13K(P<0.01).After DDP treatment,qRT-PCR results revealed that the expression of miR-214-3p was decreased,the mRNA and protein expressions of MDR1were greatly increased,and the protein levels of p62were increased in C13Kand OV2008 cells,compared to the blank control C13Kand OV2008cells(all P<0.01).Compared with the blank control C13Kand OV2008 cells,the IC50of DDP was decreased after pretreatment with OME(150μmol/L).The sensitivity of C13Kand OV2008cells to DDP was increased after OME(150μmol/L)pretreatment,the relative expression of miR-214-3p was significantly increased,the expression of MDR1protein and mRNA was decreased,and the expression of p62protein was decreased(all P<0.05).Conclusion OME pretreatment might enhance the sensitivity of ovarian can

关 键 词：miR-214-3p 自噬 奥美拉唑 卵巢癌 顺铂耐药 

分 类 号：R737.31[医药卫生—肿瘤]