miR-27a-3p靶向调节突触膜胞吐2调控宫颈癌细胞增殖凋亡的机制研究  

Mechanism of miR-27a-3p regulating proliferation and apoptosis of cervical cancer cells by targeting RIMS2

作　　者：邱娜[1] 潘长清[1] 郑茜文[1] 赵菊惠 王丹[1] QIU Na;PAN Changqing;ZHENG Qianwen;ZHAO Juhui;WANG Dan(Department of Gynaecolo-gy and Obstetrics,Mianyang Central Hospital,Mianyang 621000,Sichuan,China)

机构地区：[1]绵阳市中心医院妇产科,四川绵阳621000

出　　处：《中国性科学》2022年第1期89-93,共5页Chinese Journal of Human Sexuality

基　　金：四川省卫生和计划生育科研课题资助项目(16PJ185)。

摘　　要：目的观察微小RNA-27a-3p(miR-27a-3p)、调节突触膜胞吐2(RIMS2)对宫颈癌细胞增殖、凋亡的影响,并探究其作用机制。方法选取2014年1月至2016年6月绵阳市中心医院收治的145例行手术切除的宫颈癌患者的癌组织及癌旁组织作为研究对象。运用实时荧光定量聚合酶链反应(RT-qPCR)法检测宫颈癌组织、癌旁组织、Ect1/E6E7、HeLa细胞中miR-27a-3p、RIMS2的mRNA表达;Kaplan-Meier法绘制宫颈癌患者的生存曲线;脂质体法将anti-miR-con、anti-miR-27a-3p、pcDNA、pcDNA-RIMS2、anti-miR-27a-3p+si-con和anti-miR-27a-3p+si-RIMS2转染至HeLa细胞。细胞计数试剂盒(CCK-8)、膜联蛋白V-异硫氰酸荧光素-碘化丙锭(Annexin V-FITC/PI)双染法、蛋白免疫印迹(WB)实验检测细胞的增殖率、细胞凋亡率和RIMS2、激活的半胱氨酸天冬氨酸蛋白酶(c-Caspase)-3、c-Caspase-9的蛋白表达。双荧光素酶报告实验检测细胞的荧光活性。结果与癌旁组织或Ect1/E6E7细胞相比,癌组织、HeLa细胞中miR-27a-3p的表达显著升高,RIMS2的表达均显著降低,miR-27a-3p低表达患者的生存率显著升高(P<0.05)。抑制miR-27a-3p的HeLa细胞增殖率显著降低,凋亡率显著升高,c-Caspase-3、c-Caspase-9蛋白表达显著升高。miR-27a-3p靶向RIMS2,过表达RIMS2具有与抑制miR-27a-3p相似的功能。敲减RIMS2部分逆转了抑制miR-27a-3p对HeLa细胞的增殖抑制和凋亡促进作用。结论 miR-27a-3p促进HeLa细胞增殖,抑制凋亡,其机制与靶向RIMS2有关。Objective To observe the effects of miR-27 a-3 p and regulating synaptic membrane exocytosis 2(RIMS2) on the proliferation and apoptosis of cervical cancer cells and explore its mechanism. Methods The cancer tissues and adjacent tissues of 145 patients with cervical cancer who underwent surgical resection in Mianyang Central Hospital from January 2014 to June 2016 were selected as the research objects. Real time quantitative reverse transcription polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression of miR-27 a-3 p and RIMS2 in cervical cancer tissues, adjacent tissues, Ect1/E6 E7, HeLa cells. Kaplan-Meier method was used to draw the survival curve of patients with cervical cancer. Anti-miR-con, anti-miR-27 a-3 p, pcDNA, pcDNA-RIMS2, anti-miR-27 a-3 p+si-con, anti-miR-27 a-3 p+si-RIMS2 were all transfected into HeLa cells by liposome method. Cell Counting Kit(CCK-8), Annexin V-fluorescein isothiocyanate propidium iodide(Annexin V-FITC/PI) double staining method, Western blot(WB) test were used to detect the cell proliferation rate, apoptosis rate and protein expression of RIMS2, c-Caspase-3 and c-Caspase-9. Double luciferase reporter assay was used to detect the fluorescence activity of the cells. Results The expression of miR-27 a-3 p in cervical cancer tissues and cells was significantly higher than that in adjacent tissues or Ect1/E6 E7 cells, while the expression of RIMS2 mRNA and protein was significantly lower, and the survival rate of patients with low expression of miR-27 a-3 p was significantly higher(P<0.05). Inhibition of miR-27 a-3 p significantly reduced the proliferation rate, increased the apoptosis rate and the expression of c-Caspase-3 and c-Caspase-9 of HeLa cells. miR-27 a-3 p targeted RIMS2, overexpression RIMS2 had the same function as inhibition of miR-27 a-3 p. Knock-down RIMS2 reversed the inhibitory effect of miR-27 a-3 p on proliferation and apoptosis in HeLa cell. Conclusions miR-27 a-3 p can promote the proliferation and inhibit apoptosis of cervical cancer cells, a

关 键 词：宫颈癌 微小RNA-27a-3p RIMS2 靶向 

分 类 号：R711[医药卫生—妇产科学]