摘要
目的探讨GNAS-AS1靶向miR-204-5p对宫颈癌SiHa细胞活性凋亡和迁移的影响及分子机制。方法选取本院2016年5月至2020年5月期间收治的35例宫颈癌患者癌组织及癌旁组织;宫颈癌细胞SiHa分为si-NC组、si-GNAS-AS1组、si-GNAS-AS1+anti-miR-204-5p组、si-GNAS-AS1+anti-miR-NC组。实时荧光定量PCR(RT-qPCR)检测GNAS-AS1和miR-204-5p的表达水平;噻唑蓝(MTT)检测细胞增殖抑制率;细胞划痕实验检测细胞划痕愈合率;流式细胞术检测细胞凋亡;双荧光素酶报告实验验证GNAS-AS1和miR-204-5p的靶向关系;蛋白质印迹(Western blot)法检测蛋白表达。结果宫颈癌组织中GNAS-AS1表达水平显著升高,miR-204-5p表达水平显著降低(P<0.05)。转染GNAS-AS1干扰表达载体后,miR-204-5p表达水平升高,SiHa细胞增殖抑制率升高,SiHa细胞划痕愈合率降低,SiHa细胞凋亡率升高,SiHa细胞中Bax表达水平升高,Bcl-2表达水平降低,差异均具有统计学意义(P<0.05)。GNAS-AS1靶向调控miR-204-5p,同时抑制miR-204-5p和GNAS-AS1表达,SiHa细胞增殖抑制率降低,SiHa细胞划痕愈合率升高,SiHa细胞凋亡率降低,差异均具有统计学意义(P<0.05)。结论干扰GNAS-AS1可通过调控miR-204-5p抑制宫颈癌SiHa细胞增殖和迁移,促进细胞凋亡。
Objective To explore the effect of GNAS-AS1 targeting miR-204-5p on the apoptosis and migration of cervical cancer SiHa cells and its molecular mechanism. Methods 35 cases of cervical cancer patients with cancer tissues and adjacent tissues from May 2016 to May 2020 in our hospital were selected. Cervical cancer SiHa cells were divided into si-NC group, si-GNAS-AS1 group, si-GNAS-AS1+anti-miR-204-5p group, si-GNAS-AS1+anti-miR-NC group. Real-time fluorescent quantitative PCR(RT-qPCR) was used to detect the expression levels of GNAS-AS1 and miR-204-5p. Thiazole blue(MTT) was used to detect cell proliferation inhibition rate. Cell scratch test was to detect cell scratch healing rate. Flow cytometry was to detect cell apoptosis. The dual luciferase report experiment verified the targeting relationship between GNAS-AS1 and miR-204-5p. Western blot was used to detect protein expression. Results The expression level of GNAS-AS1 in cervical cancer tissue increased, while the expression level of miR-204-5p decreased significantly(P<0.05). After transfection of GNAS-AS1 interference expression vector, the expression of miR-204-5p increased, the inhibition rate of SiHa cells proliferation increased, the rate of SiHa cells scratch healing decreased, the apoptosis rate of SiHa cells increased, and the expression of Bax in SiHa cells increased, Bcl-2 expression decreased significantly(P<0.05). GNAS-AS1 can target and regulates miR-204-5p, and the expression of miR-204-5p and GNAS-AS1 were inhibited at the same time. And the inhibition rate of SiHa cells proliferation decreased, the rate of SiHa cells scratch healing increased, and the apoptosis rate of SiHa cells decreased significantly(P<0.05). Conclusion Interference with GNAS-AS1 can inhibit the proliferation and migration of cervical cancer SiHa cells and promote cell apoptosis by regulating miR-204-5p.
作者
晁亚萍
乔世萌
华金凤
CHAO Yaping;QIAO Shimeng;HUA Jinfeng(Department of Gynecology,Qinghai Red Cross Hospital,Xining,Qinghai 810000,China;Women’s Accounting Center of Tongde County,Qinghai Province,Tongde,Qinghai 813299,China)
出处
《中国优生与遗传杂志》
2021年第9期1208-1212,共5页
Chinese Journal of Birth Health & Heredity
