miR-377-5p通过下调VEGF-C对结肠癌的血管和淋巴管生成的影响研究

目的探讨miR-377-5p通过下调VEGF-C对结肠癌的血管和淋巴管生成的影响研究。方法该实验分为3组,miR-377-5p干扰组、miR-377-5p阴性对照组和miR-377-5p过表达组。采用实时荧光RT-PCR、Western blot、CCK-8、流式细胞、细胞划痕、Transwell、血管生成实验检测miR-377-5p m RNA和蛋白表达、细胞增殖、凋亡、迁移、侵袭、血管和淋巴管生成。结果与正常结肠组织对比,结肠癌组织中miR-377-5p m RNA和miR-377-5p蛋白的表达明显低于正常结肠组织(均P<0.05);与miR-377-5p干扰组相比,miR-377-5p阴性对照组和miR-377-5p过表达组中的m RNA及蛋白显著升高(均P<0.05),与miR-377-5p阴性对照组相比,miR-377-5p过表达组的m RNA及蛋白明显增加(P<0.05);与miR-377-5p干扰组相比,miR-377-5p阴性对照组和miR-377-5p过表达组的结肠癌细胞增殖能力明显降低(P<0.05),与miR-377-5p阴性对照组相比,miR-377-5p过表达组的结肠癌细胞增殖能力显著降低(P<0.05);与miR-377-5p干扰组相比,miR-377-5p阴性对照组和miR-377-5p过表达组的结肠癌细胞增殖凋亡明显升高(P<0.05),与miR-377-5p阴性对照组相比,miR-377-5p过表达组的结肠癌细胞凋亡能力显著升高(P<0.05);伤口愈合实验显示,miR-377-5p过表达组的结肠癌细胞的迁移能力显著低于miR-377-5p干扰组和miR-377-5p阴性对照组(P<0.05);miR-377-5p过表达组的结肠癌细胞的侵袭能力也明显低于miR-377-5p干扰组和miR-377-5p阴性对照组(P <0.05);miR-377-5p过表达组的VEGF-A、VEGF-C和P-Akt、VEGFR-3蛋白水平显著低于miR-377-5p干扰组和miR-377-5p阴性对照组(P<0.05);miR-377-5p过表达组的MVD(微血管密度)和LMVD(淋巴微血管密度)显著低于miR-377-5p干扰组和miR-377-5p阴性对照组(P<0.05)。结论 miR-377-5p在结肠癌中低表达,抑制了结肠癌细胞的增殖、细胞迁移和侵袭,促进了结肠癌细胞的凋亡能力,并直接靶向VEGF-C抑制肿瘤的血管和淋巴管生成。

Effect of miR-377-5p on angiogenesis and lymphangiogenesis in colon cancer by downregulating VEGF-C

Objective To investigate the effect of miR-377-5 p on angiogenesis and lymphangiogenesis in colon cancer by downregulating vascular endothelial growth factor(VEGF)-C.Methods Subjects were divided into the miR-377-5 p interference,miR-377-5 p negative control and miR-377-5 p overexpression groups.Real-time fluorescence RTPCR,Western blot,CCK-8,flow cytometry,cell scratches,Transwell assays,and angiogenesis experiments were used to detect miR-377-5 p m RNA and protein expression,cell proliferation,apoptosis,migration,invasion,angiogenesis and lymphangiogenesis.Results miR-377-5 p m RNA and protein expressions in the colon cancer tissue were significantly lower than those in the normal colon tissue(both P<0.05).Compared with those of the miR-377-5 p interference group,the m RNA and protein expressions in the miR-377-5 p negative control and overexpression groups were significantly increased(both P<0.05).Compared with the miR-377-5 p negative control group,miR-377-5 p overexpression colon cancer cell proliferation in the miR-377-5 p negative control and miR-377-5 p overexpression groups was significantly reduced compared with that of the miR-377-5 p interference group(P<0.05).Colon cancer cell proliferation in the miR-377-5 p overexpression group was significantly reduced compared with that of the miR-377-5 p negative control group(P<0.05).Colon cancer cell proliferation and apoptosis in the miR-377-5 p negative control and overexpression groups were significantly increased compared with those of the miR-377-5 p interference group(P<0.05).The apoptotic ability of the colon cancer cells in the miR-377-5 p overexpression group was significantly increased compared with that of the miR-377-5 p negative control group(P<0.05).The wound-healing tests showed that the migration ability of colon cancer cells in the miR-377-5 p overexpression group was significantly lower than that of the miR-377-5 p interference and miR-377-5 p negative control groups(P<0.05).Colon cancer cells in the miR-377-5 p overexpression group were also significantly less invasive than those of the miR-377-5 p interference and negative control groups(P<0.05).VEGF-A,VEGF-C,P-Akt,and VEGFR-3 protein levels were significantly lower in the miR-377-5 p overexpression group than in the miR-377-5 p interference and miR-377-5 p negative control groups(P<0.05).Microvessel density and lymphatic microvessel density were significantly lower in the miR-377-5 p overexpression group than in the miR-377-5 p interference and miR-377-5 p negative control groups(P<0.05).Conclusions Low miR-377-5 p expression inhibited colon cancer cell proliferation,migration and invasion;promoted colon cancer cell apoptosis;and inhibited blood vessels and lymph in the tumor by directly targeting VEGF-C tube generation.