Experimental Study
Study on the regulation of renal cancer cell proliferation and apoptosis function by long-chain noncoding RNA XIST targeting microRNA-106a-5p
Wang Shujuan, Wu Yudong, Wu Xinchao, Feng Ziyu
Published 2020-09-08
Cite as Chin J Exp Surg, 2020, 37(9): 1681-1683. DOI: 10.3760/cma.j.cn421213-20200307-01080
Abstract
ObjectiveTo investigate the effect of long-chain noncoding RNA (lncRNA) XIST targeting microRNA (miRNA, miR)-106a-5p on the proliferation and apoptosis of renal cancer cell line and its mechanism.
MethodsThe expression of XIST in ACHN, Caki-1, Caki-2, 786-O cells and HK2 cells was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). Caki-2 cells cultured in vitro were divided into Empty group (transfected with empty vector), and XIST group (transfected with pCDNA3.1 vector containing full-length XIST sequence). The expression levels of XIST and miR-106a-5p were detected by RT-qPCR, and cell proliferation, cell apoptosis were detected by methyl thiazolyl tetrazolium (MTT) method and flow cytometry. The target binding relationship between XIST and miR-106a-5p was detected by double luciferase reporter gene assay. After co-transfection of miR-106a-5p mimics and XIST vector, the effect of up-regulation of miR-106a-5p expression on the proliferation and apoptosis of Caki-2 cells which were XIST over expression was observed.
ResultsCompared with HK2 cells, the expression level of XIST in ACHN, Caki-1, Caki-2, 786-O cells decreased significantly (relative expression level was 0.516±0.031, 0.605±0.021, 0.241±0.024, 0.355±0.042, t=11.666, 11.321, 20.321, 12.992, P<0.05). Compared with the Empty group, the expression level of XIST (relative expression level was 175.829±1.021,t=20.471, P<0.05), cell apoptosis rate (1.304 times vs. control group,t=11.371, P<0.05) in XIST group were significantly higher, while the expression level of miR-106a-5p (relative expression level was 0.415±0.035,t=13.717, P<0.05) and cell viability (0.759 times vs. control group) were significantly lower (t=6.388, P<0.05). The double luciferase reporter gene experiment confirmed that miR-106a-5p could target with XIST. After miR-106a-5p mimics was successfully transfected to up-regulate the expression of miR-106a-5p, the inhibition of proliferation and promotion of apoptosis of Caki-2 cells by high expression of XIST were reversed.
ConclusionOver expression of XIST could inhibit the proliferation and promote the apoptosis of Caki-2 cells by targeting miR-106a-5p.
Key words:
Renal cancer; Long chain noncoding RNA XIST; Cell proliferation; Cell apoptosis; MicroRNA-106a-5p
Contributor Information
Wang Shujuan
Department of Urology, the First Affiliate Hospital of Zhengzhou University, Zhengzhou 450052, China
Wu Yudong
Department of Urology, the First Affiliate Hospital of Zhengzhou University, Zhengzhou 450052, China
Wu Xinchao
Department of Urology, the First Affiliate Hospital of Zhengzhou University, Zhengzhou 450052, China
Feng Ziyu
Department of Urology, the First Affiliate Hospital of Zhengzhou University, Zhengzhou 450052, China
