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Basic Research
lncRNA ZNF674-AS1 regulates the proliferation and migration of cervical cancer HCC94 cells via miR-510-5p/REPS2 axis
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Abstract:
Objective:To explore the role of lncRNA ZNF674-AS1 in cervical cancer and its molecular mechanism. Methods: qPCR was used to detect the expression of ZNF674-AS1 in 31 pairs of cervical cancer tissues and corresponding para-cancerous tissues that obtained from patients who had received surgical treatment during January 2019 and July 2020 in Wuhan Children’s Hospital, as well as in cervical cancer cell lines (SiHa, HeLa, C33A and HCC94) and immortalized cervical epithelial cell lines. The NF674-AS1 over-expression plasmids or the negative control plasmids were transfected into the HCC94 cell line with the least expression of ZNF674-AS1. CCK-8 method and Transwell assay were used to detect the effect of ZNF674-AS1 over-expression on the proliferation and migration of HCC94 cells.Bioinformatics methods and Dual luciferase reporter gene experiment were respectively used to predict and verify the complementary binding relationship among ZNF674-AS1, miR-510-5p and REPS2 (RALBP1 associated Eps domain containing 2). qPCR was conducted to examine the effect of ZNF674-AS1 over-expression on the levels of miR-510-5p and REPS2, and WB assay was used to detect the effect of ZNF674- AS1 over-expression on the level of molecules that related with cell proliferation and migration. Results: Compared with para-cancerous tissues, ZNF674-AS1 was significantly down-regulated in cervical cancer tissues (P<0.01). Compared with immortalized cervical epithelial cells, ZNF674-AS1 was also obviously down-regulated in cervical cancer cell lines (P<0.05 or P<0.01), with the lowest expression in HCC94 cells (P<0.01). Over-expression of ZNF674-AS1 could significantly inhibit the proliferation (P<0.05) and migration (P<0.01) of HCC94 cells. miR-510-5p was proved to interact with ZNF674-AS1, and REPS2 was proved to interact with miR-510-5p. Over-expression of ZNF674-AS1 caused a decrease in the expression level of miR-510-5p (P<0.01). but an increase in the expression level of REPS2 gene (P<0.01) in HCC94 cells, as well as the up- or down- regulation of multiple molecules (CDK2, cyclinD3, vimentin and twist) that related with cell proliferation and migration. Conclusions：lncRNA ZNF674-AS1 is low-expressed in cervical cancer tissues and cell lines and may up-regulate the expression of REPS2 gene by competitively binding miR-510-5p, thereby inhibiting the proliferation and migration of cervical cancer HCC94 cells.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No.81702561)
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