MicroRNA-486-5p通过下调Neuropilin-2的表达抑制人大肠癌中微淋巴管的生成  

Overexpression of mi R-486-5p suppresses tumor lymphangiogenesis in colorectal carcinoma by targeting neuropilin-2

作　　者：刘成霞[1] 马锋振[1] 王健[1] 李锟[1] 李琼[1] 连海峰[1] 

机构地区：[1]滨州医学院附属医院消化内科,山东省滨州市256603

出　　处：《世界华人消化杂志》2015年第27期4335-4341,共7页World Chinese Journal of Digestology

基　　金：山东省自然科学基金资助项目;No.ZR2014HQ072;山东省医药卫生科技发展计划基金资助项目;No.2014WS0490;滨州市科技发展计划基金资助项目;No.2014ZC0116~~

摘　　要：目的:探讨过表达micro RNA-486-5p(mi R-486-5p)对人大肠癌细胞系SW620体内生长的作用及其作用机制的研究.方法:构建mi R-486-5p的过表达质粒,体外脂质体法瞬时转染人大肠癌细胞SW620;建立人大肠癌裸鼠皮下移植瘤模型,实验组每3 d瘤周注射mi R-486-5p过表达质粒,对照组注射等量的空白载体.每天观察肿瘤的生长情况并每3 d测量肿瘤的体积;实时荧光定量-PCR(Real-time PCR)检测各组肿瘤组织中mi R-486-5p的表达水平;免疫组织化学技术检测移植瘤内神经纤毛蛋白-2(neuropilin-2,NRP2)及微淋巴管的的分布情况;Western blot技术检测各组移植瘤中NRP2蛋白的表达.结果:转染后,实验组中mi R-486-5p的相对表达量较对照组提高12.57±2.31倍,过表达m i R-486-5p后实验组裸鼠皮下移植瘤的生长速度较对照组明显减缓(P<0.05),瘤体质量明显减轻(P<0.05).过表达mi R-486-5p可明显抑制NRP2蛋白的表达(P<0.05),此外,mi R-486-5p组微淋巴管密度降低.结论:过表达mi R-486-5p可明显抑制大肠癌细胞的生长,抑制微淋巴管的生成,这可能与下调NRP2蛋白的表达有关.AIM：To investigate the effect of microRNA-486-5p（miRNA-486-5p） overexpression on tumor lymphangiogenesis in colorectal carcinoma cells and to explore the underlying mechanism.METHODS：A eukaryotic expression vector of miRNA-486-5p was transiently transfected into human colorectal carcinoma SW620 cells to induce miRNA-486-5p overexpression.A human colorectal carcinoma model in nude mice was then established,and the miRNA-486-5p vector was directly injected into local tumor tissue.Then,tumor size was measured every three days.The expression level of miRNA-486-5p in tumor tissue was determined by real-time PCR（RT-PCR）.Meanwhile,the expression level of Neuropilin-2（NRP2） and the number of lymphatic microvessels were detected by immunohistochemistry.The expression of NRP2 was measured by Western blot.RESULTS：The expression of miRNA-486-5p increased significantly in the miRNA-486-5transfected SW620 cells,and the mean expression level of miRNA-486-5p increased by 12.57 times ± 2.31 times after transfection.The growth speed of tumor in the miRNA-486-5p vector injected group was significantly slower than that in the control group（P〈0.05）.The mass of tumor in the miRNA-486-5p vector injected group was significantly lower than that in the control group（P〈0.05）.The expression of NRP2 and the number of lymphatic microvessels in the tumor tissue were also remarkably decreased in the miRNA-486-5p vector injected group（P〈0.05）.CONCLUSION：Overexpression of miRNA-486-5p could significantly inhibit the growth and lymphangiogenesis in colorectal carcinoma cells in vivo,and this might be related to the down-regulated expression of NRP2.

关 键 词：大肠癌 miR-486-5p 淋巴管生成 神经纤毛蛋白-2 

分 类 号：R735.34[医药卫生—肿瘤]