lncRNA SNHG16通过靶向miR-22-3p调控肾母细胞瘤WIT49细胞增殖、迁移、侵袭及凋亡的分子机制

doi:10.3969/j.issn.1005-6483.2021.09.024

摘要/Abstract

摘要： 目的探讨lncRNA SNHG16是否可通过靶向miR-22-3p调控肾母细胞瘤WIT49细胞增殖、迁移、侵袭及凋亡。
方法采用qRT-PCR法检测肾母细胞瘤组织、癌旁组织中SNHG16、miR-22-3p的表达水平；体外培养人肾母细胞瘤WIT49细胞，分别将si-NC、si-SNHG16、miR-NC、miR-22-3p mmics、si-SNHG16与anti-miR-NC、si-SNHG16与anti-miR-22-3p转染至WIT49细胞；采用qRT-PCR法检测细胞中SNHG16、miR-22-3p的表达水平；采用CCK-8法检测细胞增殖；采用Transwell小室实验检测细胞迁移及侵袭；采用流式细胞术检测细胞凋亡率；双荧光素酶报告实验检测SNHG16、miR-22-3p的靶向关系；Western blot法检测CyclinD1、MMP-2、MMP-9、Bcl-2、p21、Bax蛋白表达量。
结果与癌旁组织比较，肾母细胞瘤组织中SNHG16的表达水平显著升高(P＜0.05)，miR-22-3p的表达水平显著降低(P＜0.05)；转染si-SNHG16或转染miR-22-3p mimics可明显降低吸光度值及CyclinD1、MMP-2、MMP-9、Bcl-2蛋白水平(P＜0.05)，减少迁移及侵袭细胞数(P＜0.05)，提高凋亡率及p21、Bax蛋白水平(P＜0.05)；双荧光素酶实验证实SNHG16可靶向结合miR-22-3p；下调miR-22-3p表达可明显逆转抑制SNHG16表达对肾母细胞瘤WIT49细胞增殖、迁移侵袭和凋亡的作用。
结论抑制SNHG16表达可通过上调miR-22-3p的表达从而减弱肾母细胞瘤WIT49细胞增殖、迁移及侵袭能力，诱导细胞凋亡。

关键词: lncRNA SNHG16, miR-22-3p, 肾母细胞瘤, 增殖, 迁移, 侵袭, 凋亡

Abstract: Objective To explore whether lncRNA SNHG16 could regulate the proliferation,migration,invasion and apoptosis of Wilms’ tumor WIT49 cells by targeting miR-22-3p.
Methods qRT-PCR was used to detect the expression levels of SNHG16 and miR-22-3p in Wilms’ tumor tissues and adjacent tissues.Human Wilms’ tumor WIT49 cells were cultured in vitro,and si-NC,si-SNHG16,miR-NC,miR-22-3p mmics,si-SNHG16 and anti-miR-NC,si-SNHG16 and anti-miR-22- 3p was transfected into WIT49 cells.The qRT-PCR method was used to detect the expression levels of SNHG16 and miR-22-3p in the cells.The CCK-8 method was used to detect cell proliferation.Transwell cell experiment was used to detect cell migration and invasion.Flow cytometry was used to detect the apoptosis rate.The dual luciferase reporter experiment detects the targeting relationship of SNHG16 and miR-22-3p.Western blot method was used to detect the protein expression of CyclinD1,MMP-2,MMP-9,Bcl-2,p21,Bax.〖WT5”HZ〗Results Compared with adjacent tissues,the expression level of SNHG16 in Wilms’ tumor tissue was significantly increased(P＜0.05),and the expression level of miR-22-3p was significantly decreased(P＜0.05).Transfection of si-SNHG16 or miR-22-3p mimics could significantly reduce the OD value and the protein levels of CyclinD1,MMP-2,MMP-9 and Bcl-2(P＜0.05),and reduce the number of migration and invasion cells(P＜ 0.05),increase the apoptosis rate and the levels of p21 and Bax protein(P＜0.05).The dual luciferase report experiment confirmed that SNHG16 could target miR-22-3p.Down-regulating the expression of miR-22-3p could significantly reverse the effect of inhibiting the expression of SNHG16 on the proliferation,migration,invasion and apoptosis of Wilms tumor WIT49 cells.
Conclusion Inhibition of SNHG16 expression could reduce the proliferation,migration and invasion ability of Wilms’ tumor WIT49 cells by up-regulating the expression of miR-22-3p,and induce cell apoptosis.

Key words: lncRNA SNHG16, miR-22-3p, Wilms’ tumor, proliferation, migration, invasion, apoptosis

倪志福汪晓玲屈振繁孙恒王波. lncRNA SNHG16通过靶向miR-22-3p调控肾母细胞瘤WIT49细胞增殖、迁移、侵袭及凋亡的分子机制[J]. 临床外科杂志, 2021, 29(9): 883-888.

NI zhifu, WANG Xiaoling, QU Zhenfan, et al.. The molecular mechanism of lncRNA SNHG16 regulating the proliferation,migration,invasion and apoptosis of Wilms’ tumor WIT49 cells by targeting miR-22-3p[J]. JOURNAL OF CLINICAL SURGERY, 2021, 29(9): 883-888.

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