摘要
目的探讨白介素-29(IL-29)对结肠癌SW480细胞增殖和凋亡的影响及作用机制。方法IL-29诱导SW480细胞,四甲基噻唑蓝染色法(MTT)检测细胞增殖,流式细胞仪检测细胞凋亡,qRT-PCR检测lncRNA DLEU1和miR-149-5p表达,Western blot法检测细胞周期蛋白D1(CyclinD1)、p21、B淋巴细胞瘤-2(Bcl-2)和B淋巴细胞瘤-2相关蛋白(Bax)蛋白表达。双荧光素酶实验验证lncRNA DLEU1和miR-149-5p靶向关系。转染DLEU1小干扰RNA或miR-149-5p模拟物至SW480细胞,上述相同方法观察抑制DLEU1表达或过表达miR-149-5p对SW480细胞增殖和凋亡的影响。结果IL-29、抑制DLEU1表达或过表达miR-149-5p均可降低SW480细胞OD值、CyclinD1和Bcl-2蛋白表达(P<0.05),提高细胞凋亡率、p21和Bax蛋白表达(P<0.05)。IL-29可降低SW480细胞DLEU1表达(P<0.05),促进miR-149-5p表达(P<0.05)。DLEU1靶向负调控miR-149-5p表达。过表达DLEU1可逆转IL-29对SW480细胞增殖和凋亡的影响。结论IL-29可能通过调控DLEU1/miR-149-5p通路抑制结肠癌细胞增殖,并诱导其凋亡。
Objective To investigate the effect of IL-29 on proliferation and apoptosis of colon cancer SW480 cells and its mechanism.Methods SW480 cells were induced by IL-29,cell proliferation was detected by MTT assay,apoptosis was detected by flow cytometry,lncRNA DLEU1 and miR-149-5p were detected by qRT-PCR,and CyclinD1,p21,Bcl-2 and Bax protein expression were detected by Western blot.Dual luciferase assay verified the targeting relationship of lncRNA DLEU1 and miR-149-5p.DLEU1 small interfering RNA or miR-149-5p mimic was transfected into SW480 cells.The same methods were used to observe the effects of inhibiting DLEU1 expression or overexpressing miR-149-5p on proliferation and apoptosis of SW480 cells.Results IL-29,inhibition of DLEU1 expression or overexpression of miR-149-5p could decrease OD value,CyclinD1,and Bcl-2 protein expression in colon cancer SW480 cells(P<0.05),increase apoptosis rate,p21 and Bax protein expression(P<0.05).IL-29 could decrease the expression of DLEU1 in colon cancer SW480 cells(P<0.05)and promote the expression of miR-149-5p(P<0.05).DLEU1 targeted negative regulation of miR-149-5p expression.Overexpression of lncRNA DLEU1 reversed the effect of IL-29 on proliferation and apoptosis of colon cancer SW480 cells.Conclusion IL-29 may inhibit colon cancer cell proliferation and induce it apoptosis by regulating DLEU1/miR-149-5p pathway.
作者
刘杰
张文天
杜锦波
LIU Jie;ZHANG Wen-tian;DU Jin-bo(Department of Gastrointestinal Surgery,Cancer Hospital of Cangzhou People′s Hospital,Cangzhou 061000,China)
出处
《现代消化及介入诊疗》
2020年第1期31-36,共6页
Modern Interventional Diagnosis and Treatment in Gastroenterology
基金
沧州市科学技术研究与发展指导计划(172302047)
