circVAPA靶向miR-628-5p调控宫颈癌HeLa细胞的增殖和凋亡  

The proliferation and apoptosis regulations of cervical cancer HeLa cells by circVAPA targeting miR-628-5p

作　　者：魏杏茹 庞旭姣 吕向坤 WEI Xingru;PANG Xujiao;LYU Xiangkun(Baoding Maternal and Child Health Hospital for Gynecology,Baoding,Hebei 071000,China;Obstetrics and Gynecology of the 7th People’s Hospital of Hebei Province,Dingzhou,Hebei 073000,China;Obstetrics and Gynecology,Hospital of the 82nd Group Army of PLA,Baoding,Hebei 071000,China)

机构地区：[1]保定市妇幼保健院妇科,河北保定071000 [2]河北省第七人民医院妇产科,河北定州073000 [3]中国人民解放军陆军第八十二集团军医院妇产科,河北保定071000

出　　处：《中国优生与遗传杂志》2022年第3期391-396,共6页Chinese Journal of Birth Health & Heredity

摘　　要：目的 探讨环状RNA(circRNA)囊泡相关膜蛋白相关蛋白A(circVAPA)对宫颈癌细胞HeLa增殖、凋亡的影响及分子机制。方法 实时定量PCR(RT-qPCR)检测宫颈癌组织和癌旁组织中circVAPA和miR-628-5p表达。生物信息学分析、双荧光素酶报告基因实验和RT-qPCR用于阐明circVAPA和miR-628-5p潜在相关性。脂质体法将circVAPA小干扰RNA(si-circVAPA)、miR-628-5p模拟物分别转染HeLa,运用四甲基偶氮唑蓝(MTT)法、流式细胞术检测干扰circVAPA或过表达miR-628-5p对HeLa细胞增殖和凋亡的影响。免疫印迹法(Western blot)检测Ki-67、B细胞淋巴瘤2(Bcl-2)和Bcl相关X蛋白(Bax)表达水平。结果 宫颈癌组织中circVAPA表达较癌旁组织显著升高,而miR-628-5p表达较癌旁组织显著降低(P<0.05)。miR-424-5p与LINC00210直接结合,干扰LINC00210后miR-424-5p表达水平显著升高,过表达LINC00210后miR-424-5p表达水平显著降低(P<0.05)。干扰circVAPA或过表达miR-628-5p后,HeLa细胞活力、Ki-67和Bcl-2蛋白表达显著降低,细胞凋亡率、Bax蛋白表达显著升高(P<0.05)。抑制miR-628-5p表达能够显著逆转干扰circVAPA对HeLa细胞增殖、凋亡以及相关蛋白表达的影响(P<0.05)。结论 宫颈癌中circVAPA呈高表达,干扰circVAPA通过靶向miR-628-5p能够抑制宫颈癌HeLa细胞增殖并诱导细胞凋亡。Objective To investigate the effect of circular RNA(circRNA) vesicle-associated membrane protein-associated protein A(circVAPA) on the proliferation and apoptosis of cervical cancer cells HeLa,and further to explore its molecular mechanism.Methods The expression of circVAPA and miR-628-5p in cervical cancer tissues and adjacent tissues was measured by real-time quantitative PCR(RT-qPCR).Bioinformatics,dual luciferase report experiment and RT-qPCR were performed to clarify the potential correlation between circVAPA and miR-628-5p.The small interfering circVAPA(si-circVAPA) and miR-628-5p mimics were transfected into HeLa by liposome method,and the effect of interfering with circVAPA or overexpressing miR-628-5p on proliferation and apoptosis of HeLa cells was detected by the methyl thiazolyl tetrazolium(MTT) and flow cytometry.The expression levels of Ki-67,B-cell lymphoma 2(Bcl-2) and Bcl-related X protein(Bax) were detected by Western blot.Results The expression of circVAPA in cervical cancer tissues was significantly higher than that in adjacent tissues,while the expression of miR-628-5p was significantly lower than that in adjacent tissues(P<0.05).miR-424-5p directly bound to LINC00210,and the expression level of miR-424-5p increased significantly after interfering with LINC00210,while the expression level of miR-424-5p decreased significantly after LINC00210 overexpression(P<0.05).After interfering with circVAPA or overexpressing miR-628-5p,cell viability,expression of Ki-67 and Bcl-2 proteins significantly reduced,whereas apoptosis rate and Bax protein expression significantly increased(P<0.05).miR-628-5p inhibition could reverse the effect of interfering with circVAPA on the proliferation,apoptosis and related protein expression HeLa cells(P<0.05).Conclusion circVAPA highly expressed in cervical cancer.Interfering with circVAPA can inhibit the proliferation and induce apoptosis of cervical cancer HeLa cells by targeting miR-628-5p.

关 键 词：环状RNA囊泡相关膜蛋白相关蛋白A 宫颈癌 增殖 凋亡 miR-628-5p 

分 类 号：R737.33[医药卫生—肿瘤]