宫颈癌组织中circDENND4C的表达对宫颈癌HeLa细胞增殖、迁移和侵袭的影响  

Effects of circDENND4C in cervical cancer tissues on the proliferation, migration and invasion of cervical cancer HeLa cells

作　　者：闫雪[1] 李春梅[2] 陈燕娥[2] 王圣坦 YAN Xue;LI Chunmei;CHEN Yan'e;WANG Shengtan(Department of Traditional Chinese Medicine,Haikou Hospital of the Maternal and Child Health,Haikou,Hainan 570203,China;Department of Gynecology,Haikou Hospital of the Maternal and Child Health,Haikou,Hainan 570203,China)

机构地区：[1]海口市妇幼保健院中医科,海南海口570203 [2]海口市妇幼保健院妇科,海南海口570203

出　　处：《中国优生与遗传杂志》2022年第4期609-614,共6页Chinese Journal of Birth Health & Heredity

基　　金：海南省自然科学基金面上项目(821MS133)。

摘　　要：目的 探讨环状RNADENN/MADD域内含蛋白4C(circDENND4C)在宫颈癌组织中的表达及其对宫颈癌HeLa细胞增殖、迁移、侵袭的影响和机制。方法 收集41例宫颈癌患者的癌组织及其癌旁组织,实时定量聚合酶链反应(RT-qPCR)检测组织中circDENND4C和miR-4465表达。体外培养HeLa细胞,双荧光素酶报告基因实验验证circDENND4C和miR-4465的调控关系。HeLa细胞分为正常对照(NC)组、小干扰RNA阴性对照(si-NC)组、circDENND4C小干扰RNA(si-circDENND4C)组、模拟对照(miR-NC)组、miR-4465组、si-circDENND4C+抑制剂阴性序列(anti-miR-NC)组和si-circDENND4C+miR-4465抑制剂(anti-miR-4465)组,通过四噻唑蓝(MTT)法检测细胞增殖,Transwell检测细胞迁移和侵袭,Westernblot检测细胞中Ki-67、基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)蛋白表达。结果 宫颈癌组织中circDENND4C的表达显著高于癌旁组织(P<0.05),而miR-4465的表达显著低于癌旁组织(P<0.05)。circDENND4C在HeLa细胞中靶向负调控miR-4465表达。si-circDENND4C组HeLa细胞光密度(OD)值、迁移数、侵袭数及细胞中Ki-67、MMP-2和MMP-9蛋白表达量显著低于si-NC组(P<0.05)。miR-4465组HeLa细胞OD值、迁移数、侵袭数及Ki-67、MMP-2和MMP-9蛋白表达量显著低于miR-NC组(P<0.05)。si-circDENND4C+anti-miR-4465组HeLa细胞OD值、迁移数、侵袭数及Ki-67、MMP-2和MMP-9蛋白表达量显著高于si-circDENND4C+anti-miR-NC组(P<0.05)。结论 circDENND4C在宫颈癌组织中表达升高,其可通过负调控miR-4465促进宫颈癌细胞增殖、迁移和侵袭。Objective To investigate the expression of circular RNA DENN/MADD domain containing 4C(circDENND4C) in cervical cancer tissues and the mechanism and effects on the proliferation, migration and invasion of cervical cancer HeLa cells. Methods The cancerous tissues and adjacent tissues of 41 patients with cervical cancer were collected, and then real-time quantitative polymerase chain reaction(RT-qPCR) method was used to detect the expression of circDENND4C and miR-4465 in the tissues. HeLa cells were cultured in vitro, and the dual luciferase reporter gene experiment verified the regulatory relationship between circDENND4C and miR-4465. HeLa cells were divided into normal control(NC) group, si-NC group,si-circDENND4C group, miR-NC group, miR-4465 group, si-circDENND4C+anti-miR-NC group and si-circDENND4C+anti-miR-4465 group. Methylthiazoletrazolium(MTT) method was used to detect cell proliferation. Cell migration and invasion were detected by Transwell. Western blot was used to detect the expression of protein Ki-67, matrix metalloproteinase 2(MMP-2) and matrix metalloproteinase 9(MMP-9) in cells. Results The expression of circDENND4C in cervical cancer tissues was significantly higher than that in adjacent tissues(P<0.05), while the expression of miR-4465 was significantly lower than that in adjacent tissues(P<0.05). circDENND4C negatively regulated the expression of miR-4465 in HeLa cells. The optical density(OD) value, the number of migration and invasion of HeLa cells, and the protein expression of Ki-67, MMP-2and MMP-9 in the si-circDENND4C group were significantly lower than those in the si-NC group(P<0.05). The OD value,the number of migration and invasion of HeLa cells, and the protein expression of Ki-67, MMP-2 and MMP-9 in the miR-4465group were significantly lower than those in the miR-NC group(P<0.05). The OD value, the number of migration and invasion of HeLa cells, and the protein expression of Ki-67, MMP-2 and MMP-9 in the si-circDENND4C+anti-miR-4465 group were significantly higher than those in the

关 键 词：宫颈癌 circDENND4C miR-4465 细胞增殖 迁移 

分 类 号：R737.33[医药卫生—肿瘤]