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[LIM和SH3蛋白1在肾透明细胞癌中的表达及其对肾透明细胞癌786-O细胞侵袭和迁移的影响]

[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].

作者信息

Jin B, Gao L, Li W, Chen J C, Wen R M, Wang J Q

机构信息

Laboratory of Cancer, Xuzhou Medical College, Xuzhou 221002, China.

the Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2017 Mar 23;39(3):166-171. doi: 10.3760/cma.j.issn.0253-3766.2017.03.002.

Abstract

To investigate the expression of LIM and SH3 protein 1 (LASP1) in renal cell carcinoma and its significance in the invasion and migration of renal clear cell carcinoma 786-O cell line. 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 lymph node metastasis was analyzed by Western blot. Furthermore, small interfering RNA (siRNA) targeting LASP1 was constructed and transfected into 786-O cells to downregulate LASP1 expression. The interference effect of LASP1 siRNA on LASP1 protein and the expression of related proteins in epithelial mesenchymal transition (EMT) pathway were detected by Western blot. The effects of LASP1 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. 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%, =0.002). The expression of LASP1 in renal cell carcinoma was positively correlated with lymph node metastasis and TNM stage of renal cell carcinoma (<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, <0.05). The LASP1 siRNA remarkably down-regulated the expression of LASP1 protein in 786-O cells. The abilities of proliferation, invasion and migration of 786-O cells were decreased significantly in the LASP1 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) and control group (0.691±0.037, <0.05). The relative expression of N-cadherin protein in the siRNA group (0.449±0.047) was significantly lower than those in the siRNA-NC group (0.613±0.018) and control group (0.633±0.045, <0.05). The relative expression of vimentin protein in the siRNA group (0.477±0.029) was significantly lower than those in the siRNA-NC group (0.598±0.069) and control group (0.633±0.045, <0.05 for both). LASP1 is highly expressed in renal clear cell carcinoma, which is closely related to the development of the cancer. The effects of LASP1 on the invasion and migration of 786-O cells and lymph node metastasis may be related to the EMT.

摘要

探讨LIM和SH3蛋白1(LASP1)在肾细胞癌中的表达及其在肾透明细胞癌786 - O细胞系侵袭和迁移中的意义。采用免疫组织化学方法分析41例肾细胞癌组织和正常肾组织中LASP1的表达水平,并进一步分析LASP1表达水平与临床特征的关系。采用蛋白质印迹法分析10例有或无淋巴结转移的肿瘤组织中LASP1的表达。此外,构建靶向LASP1的小干扰RNA(siRNA)并转染至786 - O细胞中以下调LASP1表达。采用蛋白质印迹法检测LASP1 siRNA对LASP1蛋白及上皮间质转化(EMT)途径相关蛋白表达的干扰作用。然后分别使用CCK8法、transwell细胞迁移系统和蛋白质印迹分析评估LASP1敲低对细胞增殖、迁移、侵袭及基因表达的影响。肾透明细胞癌组织中LASP1表达的阳性率为90.2%(37/41),显著高于癌旁组织(29.3%,P = 0.002)。肾细胞癌中LASP1的表达与肾细胞癌的淋巴结转移及TNM分期呈正相关(P<0.05)。蛋白质印迹结果显示,LASP1在肾细胞癌中高表达(1.459±0.628),尤其在有淋巴结转移的病例中高表达(2.692±0.186,P<0.05),而在正常组织中表达为(0.696±0.053)。LASP1 siRNA显著下调786 - O细胞中LASP1蛋白的表达。LASP1 siRNA组中786 - O细胞的增殖、侵袭和迁移能力显著降低。siRNA组中E - cadherin蛋白的相对表达(0.848±0.020)显著高于siRNA - NC组(0.671±0.018)和对照组(0.691±0.037,P<0.05)。siRNA组中N - cadherin蛋白的相对表达(0.449±0.047)显著低于siRNA - NC组(0.613±0.018)和对照组(0.633±0.045,P<0.05)。siRNA组中波形蛋白(vimentin)蛋白的相对表达(0.477±0.029)显著低于siRNA - NC组(0.598±0.069)和对照组(0.633±0.045,两者P<0.05)。LASP1在肾透明细胞癌中高表达,这与癌症的发展密切相关。LASP1对786 - O细胞侵袭和迁移及淋巴结转移的影响可能与EMT有关。

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