LIM和SH3蛋白1蛋白在肾透明细胞癌组织中的表达及其对肾透明细胞癌786－O细胞侵袭和迁移能力的影响  

Expression of LIM and SH3 protein 1 in renal clear cell carcinoma and its effects on invasion and migration of renal clear cell carcinoma 786-O cells

作　　者：金波[1] 高亮[1] 李望[2] 陈家存[2] 温儒民[2] 王军起[2] Jin Bo Gao Liang Li Wang Chen Jiacun Wen Rumin Wang Junqi(Laboratory of Cancer, Xuzhou Medical College, Xuzhou 221002, China the Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, China)

机构地区：[1]徐州医学院肿瘤实验室,221002 [2]徐州医学院附属医院泌尿外科,221002

出　　处：《中华肿瘤杂志》2017年第3期166-171,共6页Chinese Journal of Oncology

摘　　要：目的分析LIM和SH3蛋白1（LASP1）在肾癌组织中的表达及其与肾癌患者临床病理特征的关系，探讨沉默LASP1表达对肾透明细胞癌786－O细胞侵袭和迁移能力的影响。方法应用免疫组织化学染色检测41例肾透明细胞癌患者肿瘤组织及其配对的癌旁组织中LASP1蛋白的表达水平，并分析LASP1蛋白表达与患者临床病理特征的关系。采用Western blot法检测10例伴或不伴淋巴结转移肾透明细胞癌组织中LASP1蛋白的表达差异。以小干扰RNA（siRNA）技术转染786－O细胞，采用Western blot法检测LASP1 siRNA对LASP1蛋白的干扰效果以及对上皮间质转化（EMT）通路中相关蛋白的表达影响。采用细胞计数盒8（CCK－8）法和Transwell实验检测下调LASP1蛋白表达对786－O细胞增殖、侵袭和迁移能力的影响。结果LASP1蛋白在肾透明细胞癌组织中的阳性表达率为90.2%（37/41），明显高于其在癌旁组织中的阳性表达率（29.3%，12/41; P＝0.002）。LASP1在肾癌组织中的表达与肾癌的淋巴结转移和TNM分期呈正相关（均P〈0.05）。Western blot法检测结果显示，LASP1蛋白在癌旁正常组织、无淋巴结转移肾癌组织和有淋巴结转移肾癌组织中的相对表达量分别为0.696±0.053、1.459±0.628和2.692±0.186，差异有统计学意义（P＝0.001）。沉默786－O细胞中LASP1蛋白的表达后，786－O细胞的增殖、侵袭和迁移能力均明显降低。LASP1 siRNA转染786－O细胞48 h后，siRNA转染组、阴性对照组和空白对照组786－O细胞中上皮表型标志物E－cadherin蛋白的相对表达量分别为0.848±0.020、0.671±0.018和0.691±0.037，siRNA转染组E－cadherin蛋白的表达明显增高（P〈0.05）；siRNA转染组、阴性对照组和空白对照组786－O细胞中间质表型标志物N－cadherin蛋白的相对表达量分别为0.449±0.047、0.613±0.018和0.633±0.045，siRNA转染组N－cadherin蛋白的表达明显降低（P〈0.05）；siRNA转�Objective To investigate the expression of LIM and SH3 protein 1 （LASP 1 ） in renal cell carcinoma and its significance in the invasion and migration of renal clear cell carcinoma 786-O cell line. Methods The expression level of LASP1 in 41 cases of renal cell carcinoma tissues and normal renal tissues was analyzed by immunohistochemistry. The relationship between the expression level of LASP1 and clinical characteristics was further analyzed. Expression of LASP1 in 10 cases of tumor tissues with or without lynlph node metastasis was analyzed by Western blot. Furthermore, small interfering RNA （siRNA） targeting LASP1 was constructed and transfeeted into 786-0 cells to downregulate LASP1 expression. The interference effect of LASP1 siRNA on LASPI protein and the expression of related proteins in epithelial mesenehymal transition （EMT） pathway were detected by Western blot. Tile effects of LASPI knockdown on cell proliferation, migration and invasion and gene expression were then assessed using CCK8 assay, transwell cell migration system and western blot analysis, respectively. Results The positive rate of LASP1 expression in renal clear cell carcinoma tissues was 90.2% （37/41） , which was significantly higher than that in the adjacent tissues （29.3%, P = 0.002）. The expression of LASP1 in renal cell carcinoma was positively correlated with lymph node metastasis and TNM stage of renal cell carcinoma （P 〈 0.05 ）. The results of Western blot showed that LASP1 （0.696±0.053） was highly expressed in renal cell carcinoma （ 1.459±0.628）, especially in cases with lymph node metastasis （ 2.692±0.186, P〈 0.05 ）. The LASP1 siRNA remarkably down-regulated the expression of LASP1 protein in 786-0 cells. The abilities of proliferation, invasion and migration of 786-O cells were decreased significantly in the LASP 1 siRNA groups. The relative expression of E-cadherin protein in the siRNA group （0.848± 0.020） was significantly higher than those in the siRNA-NC group （0.671±0.018�

关 键 词：肾肿瘤 LIM和SH3蛋白1 上皮间充质转化 肿瘤转移 淋巴结 

分 类 号：R737.11[医药卫生—肿瘤]