摘要
目的探讨长链非编码RNA DLX6-AS1通过靶基因miR-103a-3p调控宫颈癌SiHa细胞增殖、迁移、侵袭和凋亡的影响及其作用机制。方法培养正常宫颈细胞Ect1/E6E7和宫颈癌细胞株HeLa、SiHa、C33a、Caski。将SiHa随机分为对照(NC)组、si-con组、si-DLX6-AS1组、PCDNA组、PCDNA-DLX6-AS1组、miR-con组、miR-103a-3p组、si-DLX6-AS1+anti-miR-con组、si-DLX6-AS1+anti-miR-103a-3p组。RT-qPCR检测各组细胞中miR-103a-3p和DLX6-AS1表达水平;蛋白质印迹(Western Blot)法检测蛋白表达水平;四甲基偶氮唑盐比色法(MTT)检测细胞活性;流式细胞术检测细胞凋亡;Transwell检测细胞迁移和侵袭;双荧光素酶报告实验检测DLX6-AS1和miR-103a-3p的靶向关系。结果与正常宫颈细胞Ect1/E6E7相比,宫颈癌细胞HeLa、SiHa、C33a、Caski中DLX6-AS1表达水平升高,miR-103a-3p表达水平降低,差异均有统计学意义(P<0.05)。干扰DLX6-AS1表达或miR-103a-3p过表达后,细胞活性降低,细胞凋亡率升高,迁移、侵袭数量减少,CyclinD1、Bcl-2、MMP-2、MMP-9表达水平降低,Bax、Cleaved caspase-3表达水平升高,差异均有统计学意义(均P<0.05)。同时抑制miR-103a-3p和DLX6-AS1表达后,宫颈癌SiHa中MMP-2、MMP-9、CyclinD1、Bcl-2表达水平升高,Bax、Cleaved caspase-3表达水平降低,细胞活性升高,细胞凋亡率降低,差异均有统计学意义(均P<0.05)。DLX6-AS1靶向调控miR-103a-3p的表达。结论干扰DLX6-AS1表达可抑制宫颈癌细胞增殖、迁移、侵袭,促进细胞凋亡,其机制可能与miR-103a-3p表达有关,可能为宫颈癌的治疗提供新思路和新靶点。
Objective To investigate the effects of lncRNA DLX6-AS1 on proliferation,migration,invasion and apoptosis of cervical cancer cells and its mechanism.Methods The normal cervical cells(ect1/E6E7)and cervical cancer cell lines HeLa,SiHa,c33a and CaSki were cultured.SiHa was randomly divided into control(NC)group,Si-CON group,Si-dlx6-AS1 group,pcDNA group,pcDNA-dlx6-AS1 group,miR-CON group,miR-103a-3p group,Si-dlx6-AS1+anti-miR-CON group,Si-dlx6-AS1+anti-miR-103a-3p group.The expression levels of miR-103a-3p and dlx6-AS1 were detected by-RT-qPCR.Western blot was used to detect the protein expression.MTT assay was used to detect cell viability and flow cytometry was used to detect apoptosis.Transwell was used to detect cell migration and invasion.Double luciferase reporter assay was used to detect the targeting relationship between dlx6-AS1 and miR-103a-3p.ResultsCompared with the normal cervical cell ect1/e6e7,the expression level of dlx6-AS1 in HeLa,SiHa,c33a and CaSki increased,and the expression level of miR-103a-3p decreased,and the difference was statistically significant(P<0.05).After interfering with dlx6-AS1 expression or miR-103a-3p overexpression,the cell activity decreased,apoptosis rate increased,migration and invasion decreased,CyclinD1,BCL-2,MMP-2,MMP-9 expression decreased,Bax and cleaved caspase-3 were increased,and the difference was statistically significant(all P<0.05).After inhibiting the expression of miR-103a 3P and dlx6-AS1,the expression levels of MMP-2,MMP-9,CyclinD1 and Bcl 2 in SiHa were increased,the expression levels of Bax and cleaved caspase-3 decreased,cell activity increased,apoptosis rate decreased,and the difference was statistically significant(all P<0.05).Dlx6-AS1 targets the expression of miR-103a-3p.Conclusion Interfering the expression of DLX6-AS1 can inhibit the proliferation,migration and invasion of cervical cancer cells and promote apoptosis.The mechanism may be related to the expression of miR-103a-3p,which will provide new ideas and targets for the treatment of cervical c
作者
周倩
郝硕月
张海燕
ZHOU Qian;HAO Shuoyue;ZHANG Haiyan(Department of Obstetrics and Gynecology, Liaoyang Central Hospital, Liaoyang 111000, Liaoning, China)
出处
《西部医学》
2021年第1期44-49,共6页
Medical Journal of West China
