长链非编码 RNA TCF7 通过 HPV-18 E6 上调 DNMT1，并通过抑制 miR-155 调节宫颈癌细胞的生物学行为。

LncRNATCF7 up-regulates DNMT1 mediated by HPV-18 E6 and regulates biological behavior of cervical cancer cells by inhibiting miR-155.

机构信息

Department of Clinical Laboratory, Chengwu People's Hospital, Chengwu Hospital Affiliated to Taishan Medical College, Heze, P.R. China.

出版信息

Eur Rev Med Pharmacol Sci. 2019 Oct;23(20):8779-8787. doi: 10.26355/eurrev_201910_19272.

DOI:10.26355/eurrev_201910_19272

PMID:31696464

Abstract

OBJECTIVE

This work aimed to study the mechanism of lncRNATCF7 upregulating DNMT1 mediated by HPV-18 E6 and regulating the biological behavior of cervical cancer cells by inhibiting miR-155.

PATIENTS AND METHODS

HPV-16 E6 enhanced DNMT1 expression in cervical cancer cells, which was detected by Western blotting. The expression of miR-155 in cervical cancer was detected by qPCR, the interaction between TCF-7 and miR-155 by Dual-luciferase reporter gene. The changes in invasion ability of cervical cancer cells and the effect of miR-155 on the invasion ability of cervical cancer cells after inhibiting TCF-7 were detected by the transwell invasion assay, while changes in migration ability of cervical cancer cells and the effect of miR-155 on migration ability of cervical cancer cells after inhibiting TCF-7 were observed by the scratch assay. The effect of inhibiting TCF-7 on the tumor size and volume of cervical cancer was detected by the subcutaneous tumor formation in nude mice.

RESULTS

E6 expression was significantly inhibited by E6 siRNA. The knockdown of endogenous HPV-16 E6 markedly inhibited the expression of DNMT1; TCF-7 specifically bound to the 3' UTR of miR-155; inhibition of TCF-7 can inhibit invasion and migration of cervical cancer cells; enhanced miR-155 after the inhibition of TCF-7 can promote the invasion and migration of cervical cancer cells; compared with NC group, the tumor volume and weight of TCF-7-siRNA group tumor-bearing was significantly reduced.

CONCLUSIONS

TCF-7 plays an important role in the development of cervical cancer. TCF-7 can target miR-155 to regulate the invasion and migration of cervical cancer cells.

摘要

目的

本研究旨在探讨长链非编码 RNA TCF7 通过 HPV-18 E6 上调 DNMT1 的机制，以及通过抑制 miR-155 调节宫颈癌细胞的生物学行为。

患者和方法

通过 Western blot 检测 HPV-16 E6 增强宫颈癌细胞中 DNMT1 的表达。通过 qPCR 检测宫颈癌中 miR-155 的表达，通过双荧光素酶报告基因检测 TCF-7 与 miR-155 的相互作用。通过 Transwell 侵袭实验检测宫颈癌细胞侵袭能力的变化，以及抑制 TCF-7 后 miR-155 对宫颈癌细胞侵袭能力的影响，通过划痕实验观察宫颈癌细胞迁移能力的变化，以及抑制 TCF-7 后 miR-155 对宫颈癌细胞迁移能力的影响。通过裸鼠皮下肿瘤形成实验检测抑制 TCF-7 对宫颈癌肿瘤大小和体积的影响。

结果

E6 siRNA 显著抑制 E6 表达。内源性 HPV-16 E6 的敲低显著抑制 DNMT1 的表达；TCF-7 特异性结合 miR-155 的 3'UTR；抑制 TCF-7 可抑制宫颈癌细胞的侵袭和迁移；抑制 TCF-7 后增强的 miR-155 可促进宫颈癌细胞的侵袭和迁移；与 NC 组相比，TCF-7-siRNA 组荷瘤肿瘤体积和重量明显降低。