Regulation of Cervical Cancer Precancerous Cells Ect1/E6E7 by miR-29b
WEI Lingjia1,2, LOU Jiayan1,2, CHEN Mengjie1,2, WANG He1,2*
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. CCND2 was 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 miR29b 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 G0/G1 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.
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