Effect of silencing ANRIL on proliferation of human cervical cancer cell line HeLa

MEI Qihua; YANG Hongyan; SHI Qianyu; LI Qiong; WANG Zhuo; ZHAO Hong

doi:10.3969/j.issn.1002-266X.2017.35.004

Abstract：Objective To observe the effect of silencing ANRIL on the proliferation of human cervical cancer cell line HeLa and to explore its mechanism. Methods The HeLa cells in the logarithmic phase were divided into the observation group and the control group,which were transfected with ANRIL-siRNA and control siRNA,respectively. After 24 and 48 h transfection,the proliferation of the two groups was observed by CCK-8 method (the result was expressed by OD 450 ). The expression of ANRIL,p21,cell cycle dependent kinase 6 (CDK6)and cyclin-dependent kinase 2 (CDK2)mRNA was detected by fluorescence quantitative PCR at 48 h after transfection,and CHIP assay was used to detect H3K27me3 in p21 gene of the two groups. Results At 24 h after transfection,OD 450 of the observation group and the control group were 0. 55 ± 0. 08 and 0. 84 ± 0. 11,respectively. At 72 h,OD 450 of the observation group and the control group were 1. 21 ± 0. 12 and 1. 82 ± 0. 14,respectively. The OD 450 of the observation group was lower than that of the control group at 24 and 72 h after transfection (all P < 0. 05). The mRNA expression of ANRIL,CDK6,and CDK2 in the observation group was lower than that in the control group at 48 h after transfection (all P < 0. 05),while the expression of p21 in the observation group was higher than that in the control group (P < 0. 05). H3K27me3 level of p21 gene in the observation group was 1. 19 ± 0. 08,which was lower than that (5. 62 ± 0. 21)of the control group at 48 h (P < 0. 05). Conclusion Silencing ANRIL can inhibit the proliferation of cervical cancer HeLa cells by reducing the H3K27me3 level in p21 gene,and thus promoting the expression of p21 and inhibiting CDK6 and CDK2 mRNA expression.