LINC02418通过Wnt/β-catenin信号通路调控膀胱癌细胞增殖、凋亡的机制研究

Study on the mechanism of LINC02418 regulating the proliferation and apoptosis of bladder cancer cells through the Wnt/β-catenin signaling pathway

目的探讨长链非编码RNA(LINC02418)通过Wnt/β-连环蛋白(β-catenin)信号通路调控膀胱癌细胞增殖、凋亡的机制研究。方法选取2019年10月至2020年8月在本院确诊的37例膀胱癌患者为研究对象,收集患者的癌组织和癌旁组织。采用qRT-PCR检测膀胱癌组织和细胞中LINC02418的相对表达量。将膀胱癌细胞T24分为空白对照组(空白培养细胞)、si-NC组(转染si-NC)、si-LINC02418组(转染si-LINC02418);CCK8、克隆形成实验检测细胞增殖;流式细胞术检测细胞凋亡;Western blot检测Bax、Bcl-2、Wnt3a、β-catenin蛋白表达。Wnt/β-catenin信号通路激活剂加入转染si-LINC02418细胞中,检测通路激活剂LiCl对抑制LINC02418处理膀胱癌细胞增殖、凋亡的作用。结果与癌旁组织相比,膀胱癌组织中的LINC02418相对表达量升高[(2.81±0.45)vs.(0.99±0.18),P<0.05]。与空白对照组、si-NC组相比,si-LINC02418组细胞中的LINC02418相对表达量降低,细胞增殖活性降低,克隆细胞数减少,差异均有统计学意义(均P<0.05)。与空白对照组、si-NC组比较,si-LINC02418组细胞的凋亡率升高,Bax蛋白表达升高,Bcl-2蛋白表达降低,差异均有统计学意义(均P<0.05)。与空白对照组、si-NC组比较,si-LINC02418组细胞的Wnt3a、β-catenin蛋白表达均降低(均P<0.05);与si-LINC02418组比较,si-LINC02418+LiCl组细胞的Wnt3a、β-catenin蛋白表达均升高(均P<0.05)。与si-LINC02418组比较,si-LINC02418+LiCl细胞的增殖活性升高,克隆细胞数增多(均P<0.05)。与si-LINC02418组比较,si-LINC02418+LiCl组细胞的凋亡率降低,Bax蛋白表达降低,Bcl-2蛋白表达升高(均P<0.05)。结论LINC02418可能通过激活Wnt/β-catenin信号通路来抑制膀胱癌细胞增殖和诱导细胞凋亡。

ObjectiveTo explore the mechanism of long non-coding RNA(LINC02418)regulating the proliferation and apoptosis of bladder cancer cells through the Wnt/β-catenin signaling pathway.MethodsThirty-seven patients with bladder cancer diagnosed in our hospital from October 2019 to August 2020 were selected as the study objects,and cancer tissues and para-cancer tissues of the patients were collected.qRT-PCR was used to detect the relative expression of LINC02418 in bladder cancer tissues and cells.Bladder cancer cell T24 were divided into control group(blank cultured cells),si-NC group(transfected with si-NC),and si-LINC02418 group(transfected with si-LINC02418).CCK8,clone formation test were used to detect cell proliferation.Flow cytometry was used to detect apoptosis.Western blot was used to detect the protein expression of Bax,Bcl-2,Wnt3a,andβ-catenin.Wnt/β-catenin signaling pathway activator was added to transfected si-LINC02418 cells to detect the effect of pathway activator LiCl on inhibiting the proliferation and apoptosis of bladder cancer cells treated by LINC02418.ResultsCompared with para-cancer tissues,the relative expression of LINC02418 in bladder cancer tissues was increased[(2.81±0.45)vs.(0.99±0.18),P<0.05].Compared with blank control group and si-NC group,the relative expression of LINC02418 in cells of si-LINC02418 group was decreased,the cell proliferation activity was decreased,and the number of cloned cells was decreased,with statistical significance(all P<0.05).Compared with the blank control group and the si-NC group,the apoptosis rate of si-LINC02418 group was increased,the expression of Bax protein was increased,and the expression of Bcl-2 protein was decreased,and the differences were statistically significant(all P<0.05).Compared with blank control group and si-NC group,the expression of Wnt3a andβ-catenin protein in si-LINC02418 group were decreased(all P<0.05).Compared with si-LINC02418 group,the expression of Wnt3a andβ-catenin protein in si-LINC02418+LiCl group were increased(all P<0.05).Compared with the si-LINC02418 group,the cell proliferation activity and the number of cloned cells in the si-LINC02418+LiCl group were increased(all P<0.05).Compared with si-LINC02418 group,the apoptosis rate,the expression of Bax protein and the expression of Bcl-2 protein in si-LINC02418+LiCl group were decreased(all P<0.05).ConclusionsLINC02418 may inhibit the proliferation of bladder cancer cells and induce cell apoptosis through Wnt/β-catenin signaling pathway.