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Effect of lncRNA ANCR on proliferation and apoptosis of colorectal cancer cells and its mechanism |
Yue Yan,Tang Zhongsheng, Chen Hui |
Department of Proctology, Sichuan Mianyang 404 Hospital, Mianyang 621000, Sichuan China |
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Abstract Objective To investigate the effect of long chain noncodingRNA (lncRNA) on the biological behavior of colorectal cancer (CRC) cells and its mechanism. Methods Cancer and adjacent tissues of 50 CRC patients were collected.The expression levels of ANCR and forkhead box protein O1 (FOXO1) in tissues were detected by RT-qPCR and Pearson correlation analysis was performed. RNA pull-down and RNA immunoprecipitation (RIP) experiments were used to confirm whether ANCR could interact with FOXO1. ANCR and FOXO1 lentivirus were transfected into CRC cell line SW480 cells. The cells were divided into the control group (untransfected), NC group (transfected with lentiviral vector), ANCR group (transfected with ANCR over-expressed lentivirus), sh-ANCR group (transfected with ANCR knockdown lentivirus), FOXO1group (transfected with FOXO1 overexpressing lentivirus), sh-ANCR + sh-FOXO1group (transfected with ANCR and FOXO1 simultaneously inhibited lentivirus). RT-qPCR was used to detect the expression of FOXO1 after over-expression and inhibition of ANCR. 5-ethynyl-2′-deoxyuridine (EdU) labeling technology and flow cytometry were used to detect cell proliferation, cell cycle and apoptosis in each group. Western blot was used to detect the expression of B-cell lymphoma-2 (BCL-2), BCL2-Associated X Protein (BAX), cycle proteins cyclin D1 (cyclin D1) and Human P27 protein (P27 protein). Results Compared with adjacent tissues, the expression of ANCR in CRC tissues increased significantly, while the expression of FOXO1 decreased significantly (P<0.01). The expression levels of the two were negatively correlated (P<0.01). ANCR is able to bind and inhibit FOXO1 expression. Compared with the NC group, the number of cells in the sh-ANCR group and FOXO1group was reduced, the cells were arrested in the G0 / G1 phase, the number of apoptosis was increased, BAX and P27 were increased, and BCL-2 and cyclin D1 were reduced (P<0.05). Compared with the sh-ANCR group, the above indexes of the sh-ANCR + sh-FOXO1 group were reversed (P<0.05). Conclusion LncRNA ANCR expression is increased in CRC and regulates cell proliferation and apoptosis of CRC cells by regulating FOXO1 expression.
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