miR-496靶向调控DCUN1D1抑制宫颈癌发生发展机制研究  

miR-496 inhibits the occurrence and development of cervical cancer via targeting DCUN1D1

作　　者：方艳惠 康晓丽 朱巧英 刘婷[1] 刘胜冬 FANG Yan-hui;KANG Xiaoli;ZHU Qiaoying;LIU Ting;LIU Shengdong(Department of Gynecology,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,China)

机构地区：[1]河北医科大学第四医院妇科,石家庄050011

出　　处：《实用医学杂志》2022年第2期139-144,共6页The Journal of Practical Medicine

基　　金：河北省医学科学研究重点课题资助项目(编号:20210082)。

摘　　要：目的研究miR-496调控DCUN1D1对宫颈癌发生发展的影响,揭示miR-496在宫颈癌中的作用。方法免疫荧光和Western blot检测DCUN1D1在宫颈癌组织和癌旁正常组织中的表达水平;采用qRT-PCR检测DCUN1D1和miR-496在不同分期宫颈癌组织中的表达水平;筛选并验证miR-496的靶基因;q RT-PCR和Western blot检测miR-496 mimics对DCUN1D1表达的影响;CCK8和Transwell检测miR-496和DCUN1D1对宫颈癌HeLa细胞增殖、迁移和侵袭的影响;构建宫颈癌裸鼠移植瘤模型,检测miR-496对移植瘤生长的影响。结果 miR-496在癌组织中表达水平较癌旁组织低表达,其中宫颈癌的临床分期越高,DCUN1D1的mRNA表达水平越高,而miR-496的表达水平越低(P <0.05);miR-496能通过结合DCUN1D1的3′-UTR抑制DCUN1D1的表达,miR-496过表达细胞DCUN1D1 mRNA和蛋白表达水平明显降低(P <0.05);相比对照组,miR-496组能抑制HeLa细胞的增殖、迁移和侵袭能力,但相比miR-496组,miR-496+DCUN1D1组细胞的迁移和侵袭能力增强(P <0.05);在宫颈癌裸鼠移植瘤模型中,miR-496能下调DCUN1D1并抑制移植瘤生长(P <0.05)。结论miR-496和DCUN1D1异常表达与宫颈癌发病机制相关,miR-496通过调控DCUN1D1抑制宫颈癌发生发展。Objective To explore the effects of miR-496/DCUN1D1 axis on cervical cancer development to reveal the role of miR-496 in cervical cancer. Methods Immunofluorescence and Western blot were used to detect DCUN1D1 expression in cervical cancer and normal adjacent tissues. QRT-PCR was used to detect the expression of DCUN1D1 and miR-496 in cervical cancer tissues of different stages. Target gene of miR-496 was screened and verified. q RT-PCR and Western blot were used to detect the effects of miR-496 mimics on DCUN1D1 and CCK8 assay and Transwell the effects of miR-496 and DCUN1D1 on HeLa cell proliferation,migration and invasion. A nude mouse xenograft model of cervical cancer was constructed to examine the effects of miR-496 on tumor growth. Results The expression of miR-496 was lower in the cervical cancer tissues than in the para-carcinoma tissues. The higher the clinical stage of cervical cancer,the higher the mRNA expression of DCUN1D1 and the lower the expression of miR-496(P < 0.05). miR-496 could inhibit DCUN1D1 expression via binding to 3’-UTR of DCUN1D1. Protein and mRNA expression of DCUN1D1 in the miR-496-overexpressed cells were significantly decreased(P < 0.05). The proliferation,migration and invasion of HeLa cells were more significantly suppressed in miR-496 group than in the control group. The migration and invasion of HeLa cells were more significantly enhanced in the miR-496+DCUN1D1 group than in the mir-496 group(P < 0.05). In the xenograft model of nude mice,miR-496 could down-regulate DCUN1D1 and inhibit the tumor growth(P < 0.05). Conclusion Abnormal expression of miR-496 and DCUN1D1 is related to the pathogenesis of cervical cancer,and miR-496 suppresses the occurrence and development of cervical cancer by targeting DCUN1D1.

关 键 词：miR-496 DCUN1D1 宫颈癌 

分 类 号：R737.33[医药卫生—肿瘤]