摘要
目的研究miR-378在宫颈癌组织中的表达水平,并探讨其对宫颈癌细胞增殖和侵袭的影响及作用机制。方法收集2012年1月至2016年1月在山东大学齐鲁医院妇科检查的宫颈组织标本185例。采用RT-qPCR检测miR-378在宫颈组织和细胞中的表达水平;用Western Blot检测细胞中不同癌基因ATG12、CCND1和pRb的表达水平;应用BrdU细胞增殖检测和Transwell侵袭实验检测细胞增殖和侵袭情况;运用Target Scan预测并筛选miR-378的基因靶点,并用双荧光素酶报告系统进行验证。结果miR-378在宫颈上皮内瘤变(CIN)Ⅲ病变组织和宫颈癌组织中的表达水平明显高于正常宫颈组织(F=103.091,t=9.381、8.936,均P<0.05);miR-378在淋巴结转移阳性的宫颈癌组织中的表达水平明显高于淋巴结转移阴性的宫颈癌组织(t=1.007,P<0.01)。miR-378的过表达显著促进了C-33A细胞的迁移和侵袭(t=5.285,P<0.05),而miR-378的低表达则显著抑制HeLa细胞的迁移和侵袭能力(t=2.941,P<0.05)。C-33A细胞中miR-378的过表达显著降低了ATG12、CCND1和pRb的表达水平(t=1.382、1.431、2.086,均P<0.05);HeLa细胞中miR-378的低表达可增加ATG12、CCND1和pRb的表达水平(t=3.961、3.062、2.894,均P<0.05)。结论miR-378能显著促进宫颈癌的转移,ATG12作为miR-378的直接作用靶点,可为宫颈癌患者病理学和治疗靶点的分子机制提供新的见解。
Objective To detect the expression of miR-378 in cervical cancer and investigate its effects on the proliferation and invasion of cancer cells as well as the underlying mechanism.Methods A total of 185 cervical tissue samples of women who received gynecological examination in Qilu Hospital of Shandong University from January 2012 to January 2016 were included in this study.Reverse transcription-quantitative polymerase chain reaction was performed to determine the expression of miR-378 in cervical tissue and C-33A cells.Western blot assay was performed to detect the expression of different cancer genes ATG12,CCND1 and pRb in C-33A cells.BrdU cell proliferation and Transwell invasion assay were performed to determine cell proliferation and invasion.Target Scan was used to predict and screen miR-378 gene targets and verified by a dual-luciferase reporter assay system.Results The expression of miR-378 in cervical intraepithelial neoplasia(CIN)III lesioned tissue and cervical cancer tissue was significantly higher than that in normal cervical tissues(F=103.091,t=9.381,8.936,both P<0.05).The expression of miR-378 in cervical cancer tissues with positive lymph node metastasis was significantly higher than that in cervical cancer tissues with negative lymph node metastasis(t=1.007,P<0.01).The overexpression of miR-378 in cervical cancer tissues significantly promoted the migration and invasion of C-33A cells(t=5.285,P<0.05),while low expression of miR-378 in cervical cancer tissues significantly inhibited the migration and invasion of HeLa cells(t=2.941,P<0.05).The overexpression of miR-378 in C-33A cells significantly decreased the expression of ATG12,CCND1and pRb(t=1.382,1.431 and 2.086,all P<0.05).The low expression of miR-378 in C-33A cells significantly increased the expression of ATG12,CCND1 and pRb(t=3.961,3.062 and 2.894,all P<0.05).Conclusion miR-378 can greatly promote the metastasis of cervical cancer cells.ATG12,as a direct target of miR-378,provides new insights into the molecular mechanism underlying cervical cancer pathology and therapeutic target.
作者
周超
谭冬梅
张友忠
韩赛
张英姿
Zhou Chao;Tan Dongmei;Zhang Youzhong;Han Sai;Zhang Yingzi(Department of Obstetrics and Gynecology,Binzhou Medical University Hospital,Binzhou 256603,Shandong Province,China;Department of Traditional Chinese Medicine,Shandong Maternal and Child Health Hospital,Jinan 250014,Shandong Province,China;Deoartment of Gynecology,Qilu Hospital of Shandong University,Jinan 250012,Shandong Province,China)
出处
《中国基层医药》
CAS
2021年第11期1636-1642,共7页
Chinese Journal of Primary Medicine and Pharmacy
基金
国家自然科学基金资助项目(81572559)
滨州医学院科技计划项目(BY2018KJ04)。
关键词
宫颈肿瘤
宫颈上皮内瘤样病变
微小RNA
自噬相关基因
乳头瘤病毒
细胞系
肿瘤
肿瘤转移
Uterine cervical neoplasms
Cervical intraepithelial neoplasia
MicroRNA
Autophagy related genes
Human papillomavirus
Cell line,tumor
Neoplasm metastasis
