miR-29b对宫颈癌前细胞Ect1/E6E7的调控  

Regulation of Cervical Cancer Precancerous Cells Ect1/E6E7 by miR-29b

作　　者：韦凌嘉 楼佳燕 陈梦捷 王鹤[1,2] WEI Lingjia;LOU Jiayan;CHEN Mengjie;WANG He(Guangxi Medical University Cancer Hospital,Nanning 530021,China;Guangxi Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor,Nanning 530021,China)

机构地区：[1]广西医科大学附属肿瘤医院,南宁530021 [2]广西区域性高发肿瘤早期防治研究重点实验室,南宁530021

出　　处：《中国细胞生物学学报》2023年第2期185-192,共8页Chinese Journal of Cell Biology

基　　金：广西自然科学基金(批准号:2020GXNSFAA159032);国家自然科学基金(批准号:82160443);区域性高发肿瘤重点实验室专项资金(批准号:02402215010D);广西医疗卫生适宜技术开发与推广应用项目(批准号:S2018031)资助的课题。

摘　　要：该文旨在研究miR-29b对宫颈癌前细胞Ect1/E6E7增殖、凋亡、细胞周期的调控。先通过生物信息学分析筛选出关键基因,再通过关键基因找出与其相关的miRNAs。用miR-29b上调慢病毒和miR-29b下调慢病毒转染Ect1/E6E7细胞,实时荧光定量PCR法检测转染后Ect1/E6E7细胞中miR-29b的表达量,CCK-8法检测转染后各组细胞的增殖情况,流式细胞术检测转染后各组细胞凋亡及细胞周期情况,双荧光素酶实验验证miR-29b与CCND2的靶向关系。经生物信息学分析筛选出关键基因CCND2,再由CCND2找到miR-29b。用miR-29b上下调慢病毒转染Ect1/E6E7细胞,miR-29b上调组与NC组相比,细胞增殖速率减慢,凋亡率升高(P<0.05)。miR-29b上调组与NC组和miR-29b下调组相比,G_(0)/G_(1)期细胞比例高,S期细胞比例低。miR-29b下调组与NC组比较,细胞增殖速率加快,凋亡率降低(P<0.05)。双荧光素酶实验表明,miR-29b可与CCND2靶向结合。总之,miR-29b可与CCND2靶向结合,并且可调控宫颈癌前细胞Ect1/E6E7增殖、细胞周期和细胞凋亡从而调控宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)的发生和进展。但其是否通过CCND2来实现该调控,仍需进行下一步研究。This study is aimed to explore the regulation of miR-29b on proliferation,apoptosis and cell cycle of Ect1/E6E7 cervical precancerous cells.First,the key genes were screened through bioinformatics,and then the miRNAs related to the key genes were identified.Ect1/E6E7 cells were transfected with overexpressing lentivirus and knock-down lentivirus of miR-29b,and the expression of miR-29b in the transfected Ect1/E6E7 cells was detected by real-time quantitative PCR.The proliferation of the transfected cells was detected by CCK-8,and the apoptosis and cell cycle of the transfected cells were detected by flow cytometry.CCND2was identified as the target gene of miR-29b by double luciferase assay.The key gene CCND2 was screened by bioinformatics and miR-29b was found by CCND2.Ect1/E6E7 cells were transfected with overexpressing lentivirus and knock-down lentivirus of miR-29b.Compared with NC group,the proliferation rate of miR-29b up-regulated group was slower and the apoptosis rate was higher(P<0.05).Compared with NC group and miR-29b down-regulated group,the proportion of G_(0)/G_(1) phase cells was higher and the proportion of S phase cells was lower in the miR-29b up-regulated group.Compared with NC group,the proliferation rate of miR-29b down-regulated group was faster and the apoptosis rate was lower(P<0.05).Dual luciferase assay confirmed that miR-29b could bind to CCND2.In conclusion,miR-29b can target CCND2 and regulate the proliferation,cell cycle and apoptosis of cervical precancerous cells Ect1/E6E7 to regulate the occurrence and progression of CIN(cervical intraepithelial neoplasia).But whether miR-29b achieves this regulation through CCND2 still needs to be further studied.

关 键 词：miR-29b CCND2 CIN 宫颈癌前细胞 

分 类 号：R737.33[医药卫生—肿瘤]