上调结肠癌间充质干细胞中miR-93-5p表达抑制结肠癌细胞增殖及促进凋亡的机制探讨

The mechanism of up-regulating miR-93-5p expression in colon cancer mesenchymal stem cell inhibit colon cancer cell proliferation and promote apoptosis

目的:观察miR-93-5p上调的结肠癌间充质干细胞(colon cancer mesenchymal stem cell,CCMSC)对结肠癌细胞增殖、凋亡的影响,并初步探讨其机制。方法:以贴壁细胞培养法抽提癌组织CCMSC,从正常结肠组织抽提结肠间充质干细胞(colon mesenchymal stem cell,CMSC),观察miR-93-5p及PD-L1的表达差异;以拟似剂上调CCMSC中miR-93-5p表达,CCK8法及流式细胞术检测CCMSC诱导的SW480细胞增殖及凋亡变化。结果:miR-93-5p在结肠癌细胞株中表达低于正常结肠上皮细胞(P<0.001),PD-L1 mRNA和PD-L1蛋白在结肠癌细胞株中表达高于正常结肠上皮细胞(P<0.001,P<0.001);miR-93-5p在CCMSC组中表达明显低于CMSC(P<0.001);与SW480细胞比较,给予CCMSC干预后SW480细胞存活率上升(P<0.001),凋亡率下降(P<0.05),与SW480细胞或CCMSC/SW480细胞比较,给予转染miR-93-5pmimics CCMSC干预的SW480细胞存活率下降(P<0.01,P<0.001),凋亡率上升(P<0.05,P<0.01)。结论:上调CCMSC中miR-93-5p通过靶向调控PD-L1抑制结肠癌细胞增殖,促进凋亡。

Objective:To observe the effect of up-regulating miR-93-5 p expression in colon cancer mesenchymal stem cell(CCMSC) on proliferation,apoptosis and discuss the mechanisms.Methods:CCMSC and colon mesenchymal stem cell(CMSC) were extracted from colon cancer or colon tissues by tissue adhere cell culture assay.miR-93-5 p and PD-L1 expression were detected.CCMSC was transfected with miR-93-5 p mimics,CCK8 and flow cytometry assay were used to detect proliferation or apoptosis.Results:miR-93-5 p in colon cancer cells were lower than in normal colon epithelial cells(P<0.001),PD-L1 mRNA and PD-L1 protein in colon cancer cells were higher than in normal colon epithelial cells(P<0.001,P<0.001).miR-93-5 p in CCMSC group was lower than in CMSC(P<0.001).Compared with SW480 cell,survival rate of SW480 was increased once interfered with CCMSC(P<0.001),apoptosis rate was decreased(P<0.05).Compared with SW480 or CCMSC/SW480,the survival rate of miR-93-5 p mimics CCMSC/SW480 was decreased(P<0.01,P<0.001),and the apoptosis rate was increased(P<0.05,P<0.01).Conclusion:Up-regulating miR-93-5 p expression suppresses CCMSC induced colon cancer cell proliferation,promotes apoptosis.