LINC00599靶向miR-210对直肠癌细胞增殖、凋亡的影响

Effects of LINC00599 on the proliferation and apoptosis of rectal cancer cells by targeting miR-210

目的探讨LINC00599对直肠癌细胞增殖、凋亡的影响和分子机制。方法实时荧光定量PCR(RT-qPCR)直肠癌组织、癌旁组织中LINC00599的表达水平。双荧光素酶报告基因实验验证LINC00599对miR-210的靶向作用。将直肠癌细胞SW480分为pcDNA3.1、pcDNA3.1-LINC00599、anti-miR-NC、anti-miR-210、pcDNA3.1-LINC00599+miR-NC、pcDNA3.1-LINC00599+miR-210组。四甲基偶氮唑蓝(MTT)和集落形成实验检测细胞增殖能力;流式细胞术检测细胞凋亡;蛋白质印记(Western blot)检测细胞增殖核抗原(Ki67)、半胱氨酸蛋白酶3(pro-caspase-3)和裂解的caspase-3(clv-caspase-3)蛋白表达。结果直肠癌组织中LINC00599表达水平较癌旁组织降低(P<0.05)。miR-210是LINC00599的靶基因。与pcDNA3.1组比较,pcDNA3.1-LINC00599组SW480细胞存活率、克隆形成数、Ki67和pro-caspase3蛋白表达、miR-210表达降低,细胞凋亡率、cl-caspase3蛋白表达升高(P<0.05)。与anti-miR-NC组比较,anti-miR-210组SW480细胞存活率、克隆形成数、Ki67和pro-caspase3蛋白表达降低,细胞凋亡率、cl-caspase3蛋白表达升高(P<0.05)。与pcDNA3.1-LINC00599+miR-NC组比较,pcDNA3.1-LINC00599+miR-210组SW480细胞存活率、克隆形成数、Ki67和pro-caspase3蛋白表达升高,细胞凋亡率、cl-caspase3蛋白表达降低(P<0.05)。结论直肠癌中LINC00599表达降低。过表达LINC00599通过靶向miR-210可抑制直肠癌细胞增殖,诱导细胞凋亡。

Objective To investigate the effects of LINC00599 on the proliferation and apoptosis of rectal cancer cells by targeting miR-210,and to explore its molecular mechanism.Methods The expression levels of LINC00599 in rectal cancer tissues and adjacent tissues were detected by real-time fluorescence quantitative PCR(RT-qPCR).The double luciferase reporter experiment was used to verify the targeting effects of LINC00599 on miR-210.Rectal cancer cells SW480 were divided into pcDNA3.1,pcDNA3.1-LINC00599,anti-miR-NC,anti-miR-210,pcDNA3.1-LINC00599+anti-miR-NC,pcDNA3.1-LINC00599+anti-miR-210 groups,respectively.And methyl thiazolyl tetrazolium(MTT)and colony formation experiments were used to detect cell proliferation;flow cytometry was used to detect apoptosis;Western Blot was used to detect the expressions of Ki67,pro-caspase-3 and caspase-3(clv-caspase-3)protein.Results The expression levels of LINC00599 in rectal cancer tissues were significantly lower than those in adjacent tissues(P<0.05).The miR-210 was a target gene of LINC00599.Compared with those in pcDNA3.1 group,the survival rate,colony formation number,Ki67 and pro-caspase3 protein expression,miR-210 expression of SW480 cells in pcDNA3.1-LINC00599 group were significantly decreased,and the apoptosis rate and cl-caspase3 protein expression were significantly increased(P<0.05).Compared with those in anti-miR-NC group,the survival rate,colony formation number,Ki67 and pro-caspase3 protein expression,miR-210 expression of SW480 cells were significantly decreased in anti-miR-210 group,but the apoptosis rate and cl-caspase3 protein expression were significantly increased(P<0.05).Compared with those in pcDNA3.1-LINC00599+miR-NC group,the survival rate,clone formation number,Ki67 and pro-caspase3 protein expression were significantly increased,and apoptotic rate,cl-caspase3 protein expression were significantly decreased in pcDNA3.1-LINC00599+miR-210(P<0.05).Conclusion The expression levels of LINC00599 are ecreased in rectal cancer,and overexpression of LINC00599 can inhibit rectal cancer cell proliferation and induce cell apoptosis by targeting miR-210.