摘要
目的:研究miR-874-3p是否通过靶向GALNT4影响宫颈癌细胞增殖和凋亡。方法:qRT-PCR和Western blot检测宫颈癌细胞系miR-874-3p、GALNT4 mRNA和GALNT4蛋白表达。HeLa细胞转染miR-874-3p或si-GALNT4,Western blot检测GALNT4、细胞周期蛋白D1(CyclinD1)、Cleaved-Caspase-3、β-catenin表达,CCK-8检测细胞增殖,流式细胞术检测细胞凋亡。starBase预测与双荧光素酶活性检测分析miR-874-3p和GALNT4的靶向关系。共转染miR-874-3p和pcDNA-GALNT4,观察过表达GALNT4对miR-874-3p过表达诱导的HeLa细胞增殖和凋亡的影响。结果:与宫颈上皮永生化细胞H8相比,宫颈癌SiHa、HeLa、Caski细胞miR-874-3p表达降低,GALNT4 mRNA和蛋白表达增加(P<0.05)。过表达miR-874-3p降低HeLa细胞CyclinD1、β-catenin蛋白表达及细胞增殖率,提高Cleaved-Caspase-3蛋白表达和凋亡率(P<0.05)。抑制GALNT4表达降低HeLa细胞CyclinD1蛋白表达及增殖率,提高Cleaved-Caspase-3蛋白表达和细胞凋亡率(P<0.05)。miR-874-3p靶向调控GALNT4表达。过表达GALNT4可部分反转miR-874-3p过表达对宫颈癌HeLa细胞增殖、CyclinD1、β-catenin蛋白表达的抑制作用,及对细胞凋亡、Cleaved-Caspase-3蛋白表达的促进作用。结论:miR-874-3p通过靶基因GALNT4调控Wnt/β-catenin信号通路,抑制宫颈癌细胞增殖,并诱导其凋亡。
Objective:To investigate whether miR-874-3p affects proliferation and apoptosis of cervical cancer cells by targeting GALNT4.Methods:Expressions of miR-874-3p,GALNT4 mRNA and protein in cervical cancer cell lines were detected by qRTPCR and Western blot. miR-874-3p or si-GALNT4 was transfected into HeLa cells. Western blot was used to determine GALNT4,CyclinD1,Cleaved-Caspase-3 and β-catenin protein expressions. CCK-8 was employed to measure cell proliferation. Flow cytometry was used to detect cell apoptosis. starBase prediction and detection of dual luciferase activity were used to analyze targeting relationship between miR-874-3p and GALNT4. miR-874-3p and pcDNA-GALNT4 were co-transfected,and effects of over-expression of GALNT4 on proliferation and apoptosis of HeLa cells induced by over-expression of miR-874-3p were observed.Results:Compared with cervical epithelial immortal cells H8,expression of miR-874-3p in cervical cancer cells SiHa,HeLa and Caski was decreased,while expressions of GALNT4 mRNA and protein were increased(P<0.05). Over-expression of miR-874-3p decreased CyclinD1,β-catenin protein levels and proliferation rate,but increased Cleaved-Caspase-3 protein expression and apoptosis rate in HeLa cells(P<0.05). Inhibiting expression of GALNT4 decreased CyclinD1 protein expression and proliferation rate,increased Cleaved-Caspase-3protein level and apoptotic rate in HeLa cells(P<0.05). miR-874-3p targeting regulated GALNT4 expression. Over-expression of GALNT4 could partially reverse inhibitory effect of miR-874-3p over-expression on proliferation,CyclinD1,β-catenin protein expressions and promotion of apoptosis and Cleaved-Caspase-3 protein expression in cervical cancer cell HeLa.Conclusion:miR-874-3p regulates Wnt/β-catenin signaling pathway through target gene GALNT4,inhibits proliferation of cervical cancer cells,and induces cell apoptosis.
作者
李娜
李焱
高雪梅
朱一麟
贺漪(指导)
LI Na;LI Yan;GAO Xuemei;ZHU Yilin;HE Yi(Department of Obstetrics and Gynecology,Wuhan First Hospital,Wuhan 430022,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第16期1983-1987,共5页
Chinese Journal of Immunology
