微小RNA-140-5p下调Jagged1抑制肾透明细胞癌迁移、侵袭及血管生成  

Down-regulation of Jagged1 by microRNA-140-5p inhibits migration, invasion and angiogenesis in renal clear cell carcinoma

作　　者：吴国森 徐林熙 许振强[1] 林建贵[1] 刘洪杰 庄志明[1] Wu Guosen;Xu Linxi;Xu Zhenqiang;Lin Jiangui;Liu Hongjie;Zhuang Zhiming(Department of Urology,Zhangzhou Municipal Hospital,Zhangzhou 363000,China)

机构地区：[1]福建省漳州市医院泌尿外科,363000

出　　处：《中华实验外科杂志》2022年第6期1038-1041,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨miR-140-5p介导Jagged1对肾透明细胞癌(ccRCC)迁移、侵袭及血管生成能力的影响。方法选取福建省漳州市医院泌尿外科2019年4月至2020年12月经肾癌根治术患者的肾透明细胞癌及癌旁组织标本17例, 选取购自美国菌种保藏中心肾透明细胞癌细胞系(786-0)和肾皮质近曲小管上皮细胞(HK-2), 荧光定量聚合酶链反应(RT-qPCR)检测miR-140-5p的表达。免疫组织化学检测ccRCC组织中Jagged1的表达。将786-0细胞分为miRNA阴性对照组(miR-NC组)、过表达miR-140-5p组(miR-140-5p组)、Jagged1阴性对照组(Jagged1-NC组)、敲减Jagged1组(si-Jagged1组)。使用划痕、transwell小室和血管生成实验, 分别检测细胞迁移、侵袭和血管生成的改变。RT-qPCR检测miR-140-5p过表达后Jagged1 mRNA的表达, 蛋白质印迹法(Western blot)检测Notch/Jagged1信号通路中Jagged 1蛋白的表达。计量资料比较采用t检验, 计数资料采用χ^(2)检验。结果 ccRCC组织(0.318±0.144)中miR-140-5p的表达量显著低于正常肾组织(1.028±0.033), 差异有统计学意义(t=-19.787, P<0.01);786-0肾癌细胞(0.295±0.064)中miR-140-5p的表达量显著低于HK-2正常肾细胞(1.009±0.157), 差异有统计学意义(t=-8.366, P<0.01);在体外, miR-140-5p组的细胞迁移率(0.394±0.015)、侵袭细胞数(142.000±17.874)和血管管腔样结构的形成数(46.000±5.291)显著少于miR-NC组的细胞迁移率(0.548±0.020)、侵袭细胞数(267.000±26.357)和血管管腔样结构的形成数(74.000±8.000), 差异有统计学意义(t=-10.362、-14.445、-5.056, P<0.01);与miR-140-5p相反, ccRCC组织中Jagged1的表达(92.30%)显着高于正常肾组织(36.67%), 差异有统计学意义(χ^(2)=5.356, P<0.05);si-Jagged1组的细胞迁移率(0.429±0.011)、侵袭细胞数(130.533±19.379)和血管管腔样结构的形成数(45.333±3.214, )都显著少于Jagged1-NC组的细胞迁移率(0.495±0.025)、侵袭细胞数(252.466±19.231)和血管管腔样结构的形成数(66.000±3.6Objective To investigate the effects of microRNA(miR)-140-5p mediated Jagged1 on migration,invasion and angiogenesis of renal clear cell carcinoma(ccRCC).Methods Totally,17 cases of ccRCC and renal tissue samples were selected.The ccRCC cell line(786-0)and renal cortical proximal tubular epithelial cells(HK-2)were selected and the expression of miR-140-5p was detected by fluorescence quantitative polymerase chain reaction(RT-qPCR).The expression of Jagged1 in ccRCC tissues was detected by immunohistochemistry.The 786-0 cells were divided into miRNA negative control group(miR-NC group),overexpression miR-140-5p group(miR-140-5p group),Jagged1 negative control group(Jagged1-NC group),and knockdown Jagged1 group(si-Jagged1 group).Using scratch,Transwell inserts and angiogenesis assays,alterations in cell migration,invasion and angiogenesis were detected,respectively.The expression of Jagged1 mRNA after miR-140-5p overexpression was detected by RT-qPCR,and the expression of Jagged 1 protein in the Notch/Jagged1 signaling pathway was detected by Western blotting.Measurement data were compared using the t-test,and enumeration data were analyzed using theχ^(2) test.Results The expression of miR-140-5p in ccRCC tissues(0.318±0.144)was significantly lower than that in normal renal tissues(1.028±0.033),and the difference was statistically significant(t=-19.787,P<0.01);the expression of miR-140-5p in 786-0 renal carcinoma cells(0.295±0.064)was significantly lower than that in HK-2 normal renal cells(1.009±0.157),and the difference was statistically significant(t=-8.366,P<0.01);in vitro,the cell migration rate(0.394±0.015),the number of invasive cells(142.000±17.874),and the number of vascular tube-like structure formation(46.000±5.291)in the miR-140-5p group were significantly reduced as compared with those in the miR-NC group(0.548±0.020,267.000±26.357 and 74.000±8.000 respectively),and the difference was statistically significant(t=-10.362,-14.445,-5.056,P<0.01);in contrast to miR-140-5p,the expression rate of

关 键 词：微小RNA 肾癌 迁移 侵袭 血管生成 

分 类 号：R737.11[医药卫生—肿瘤]