汪宁　　王荣江　　邵四海　　钟欢

[关键词] CML66;肾癌;细胞生物功能;信号通路

[中图分类号] R692          [文献标识码] A          [文章编号] 1673-9701（2021）21-0014-04

Effect of CML66 gene silencing on the proliferation， migration and invasion of renal cancer cells and its mechanism

WANG Ning   WANG Rongjiang   SHAO Sihai   ZHONG Huan

Department of Urology，the First Affiliated Hospital of Huzhou Normal College，Huzhou   313000， China

[Abstract] Objective To explore the effect of silencing chronic myeloid leukemia tumor antigen 66 （CML66） gene with small interfering RNA （siRNA） on the biological functions of human renal carcinoma 786-O cells and the LIS1/Dynein signaling pathway. Methods The liposome transfection method was used to transfect CML66-siRNA targeting CML66 gene into human renal carcinoma 786-O cells. After transfected CML66-siRNA was detected by Western blotting， the expression of CML66， LIS1， and Dynein protein in 786-O cells was detected. The CCK8 method was used to detect cell proliferation ability. Transwell and Matrigel methods were used to measure cell migration and invasion ability. Results Compared with the control group， the expression of CML66， LIS1， and Dynein protein in 786-O cells decreased after transfection of CML66-siRNA， and the difference was statistically significant（P<0.01）. The cell proliferation ability of the siRNA-CML66 group was significantly lower than that of the control group after 2， 3， and 4 days after transfection （P<0.05）. The ability of cell migration and invasion in the siRNA-CML66 transfection group was also significantly weaker than that in the control group （P<0.01）. Conclusion CML66 is highly expressed in human renal carcinoma 786-O cells. Down-regulation of CML66 gene can reduce the proliferation， migration， and invasion of human renal carcinoma 786-O cells. The mechanism may be related to the LIS1/Dynein signaling pathway.

[Key words] CML66; Kidney cancer; Cell biological function; Signal pathway

腎癌（Renal cell carcinoma）是较常见的泌尿系肿瘤，其发病率高且有逐年上升趋势[1-2]。由于肾癌放疗及化疗均不敏感，目前治疗方法主张根治性切除，如出现转移则预后较差[3-4]。目前对肾癌标记物的研究对指引肾癌精细化治疗具有重要作用。广谱癌基因CML66的相对分子质量66 kDa，染色体位于8q23.3。它高表达于慢性粒细胞性白血病、前列腺癌、肺癌、黑色素瘤中。而本课题组前期利用免疫组化法检测CML66蛋白在肾癌组织中也呈高表达水平[5]。本次拟研究干扰抑制CML66后对肾癌细胞的迁移、侵袭及增殖能力的影响情况，探索CML66在肾癌细胞发生发展中可能的信号通路机制，为肾癌的治疗提供可能的有效靶点，现报道如下。

1 材料与方法

1.1 材料

肾癌786-O细胞株购买于中科院上海公司;CML66、LIS1及Dynein抗体购买于Abcam公司;细胞增殖与毒性检测试剂盒购买于同仁化学研究所;穿透小室购买于康宁公司;Matrigel胶购买于Becton， Dickinson and Company公司;脂质体Lipofectamine 2000购买于Invitrogen公司。