miR-875-5p靶向基质抗原2对宫颈癌细胞迁移及侵袭行为的影响  

Effects of miR-875-5p targeting STAG2 on the migration and invasion behavior of cervical cancer cells

作　　者：何德娇[1] 梁华[2] 欧阳晓飞[2] 凌娜[1] 李小丽 梁轶岚 HE Dejiao;LIANG Hua;OUYANG Xiaofei;LING Na;LI Xiaoli;LIANG Yilan(Department of Internal Medicine-Oncology,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei,China;Department of Gynaecology,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei,China)

机构地区：[1]武汉大学人民医院肿瘤内科,武汉430060 [2]武汉大学人民医院妇科,武汉430060

出　　处：《中国性科学》2021年第11期89-93,共5页Chinese Journal of Human Sexuality

摘　　要：目的探讨miR-875-5p与基质抗原2(STAG2)的靶向关系,并分析两者对宫颈癌HeLa 229细胞侵袭及迁移的影响。方法选取2018年10月至2020年10月武汉大学人民医院收治的120例宫颈癌患者作为研究对象,留取其组织标本作为宫颈癌组。另选取同期武汉大学人民医院收治的120例子宫肌瘤患者,留取其宫颈组织标准本作为对照组。体外培养HeLa 229细胞,分为空白对照组(NG组)、阴性转染组(inhibitor-NC组)、抑制miR-875-5p表达组(miR-875-5p-inhibitor组)、共转染组(miR-875-5p-inhibitor+STAG2-siRNA组)。采用实时荧光定量聚合酶链式反应法(qRT-PCR)检测组织及各组HeLa 229细胞中miR-875-5p、STAG2 mRNA水平;划痕实验、Transwell实验分别检测各组HeLa 229细胞迁移、侵袭能力;免疫印迹法法检测STAG2蛋白及侵袭迁移相关蛋白[E-钙粘附蛋白(E-cadherin)、N-钙粘附蛋白(N-cadherin)、波形蛋白(Vimentin)]表达情况;采用TargetScan数据库预测miR-875-5p与STAG2靶向关系并用双荧光素酶报告基因实验验证。结果与对照组比较,宫颈癌组miR-875-5p水平升高,STAG2 mRNA水平降低,差异具有统计学意义(P<0.05)。与NG组、inhibitor-NC组比较,miR-875-5p-inhibitor组HeLa 229细胞miR-875-5p水平、划痕愈合率、侵袭细胞数、N-Cadherin、Vimentin蛋白表达均降低,STAG2 mRNA及其蛋白水平、E-Cadherin蛋白表达均升高,差异具有统计学意义(P<0.05)。结论抑制miR-875-5p可能靶向上调STAG2表达,抑制宫颈癌HeLa 229细胞迁移及侵袭,其有望成为宫颈癌新的潜在治疗靶点。Objective To investigate the targeting relationship between miR-875-5 p and stromal antigen 2(STAG2), and analyze their effects on the invasion and migration of cervical cancer HeLa 229 cells. Methods 120 patients with cervical cancer admitted to Renmin Hospital of Wuhan University from October 2018 to October 2020 were selected as the cervical cancer group. Another 120 patients with uterine fibroids admitted to the hospital during the same period were selected as the control group. Both group tissue specimens were collected. HeLa 229 cells were cultured in vitro, and divided into blank control group(NG group), negative transfection group(inhibitor-NC group), inhibiting miR-875-5 p expression group(miR-875-5 p-inhibitor group), and co-transfection group(miR-875-5 p-inhibitor+STAG2-siRNA group). Quantitative real-time fluorescence polymerase chain reaction(qRT-PCR) was used to detect miR-875-5 p and STAG2 mRNA levels of HeLa 229 cells in all groups. The migration and invasion abilities of HeLa 229 cells in each group were detected by scratch test and Transwell test. The expression of STAG2 protein and invasion and migration related proteins(E-cadherin, N-cadherin, Vimentin) were detected by western blotting. The targeting relationship between miR-875-5 p and STAG2 was predicted using TargetScan database and verified by dual luciferase reporter assay. Results Compared with the control group, miR-875-5 p level was increased and STAG2 mRNA level was decreased in cervical cancer group, with statistical significance(P<0.05). Compared with NG group and inhibitor-NC group, miR-875-5 p level, scratch healing rate, invasive cell number, N-cadherin and Vimentin protein expression of HeLa 229 cells in miR-875-5 p-inhibitor group were decreased. STAG2 mRNA and protein levels and E-cadherin protein expression were increased, and the differences were statistically significant(P<0.05). Conclusions Inhibition of miR-875-5 p may up-regulate STAG2 expression and inhibit the migration and invasion of HeLa 229 cells, which is expecte

关 键 词：miR-875-5p 基质抗原2 宫颈癌细胞 迁移 侵袭 

分 类 号：R711[医药卫生—妇产科学]