汪寧　　王榮江　　邵四?！　＄姎g

[關(guān)鍵詞] CML66;腎癌;細(xì)胞生物功能;信號(hào)通路

[中圖分類號(hào)] R692? ? ? ? ? [文獻(xiàn)標(biāo)識(shí)碼] A? ? ? ? ? [文章編號(hào)] 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）是較常見的泌尿系腫瘤，其發(fā)病率高且有逐年上升趨勢[1-2]。由于腎癌放療及化療均不敏感，目前治療方法主張根治性切除，如出現(xiàn)轉(zhuǎn)移則預(yù)后較差[3-4]。目前對(duì)腎癌標(biāo)記物的研究對(duì)指引腎癌精細(xì)化治療具有重要作用。廣譜癌基因CML66的相對(duì)分子質(zhì)量66 kDa，染色體位于8q23.3。它高表達(dá)于慢性粒細(xì)胞性白血病、前列腺癌、肺癌、黑色素瘤中。而本課題組前期利用免疫組化法檢測CML66蛋白在腎癌組織中也呈高表達(dá)水平[5]。本次擬研究干擾抑制CML66后對(duì)腎癌細(xì)胞的遷移、侵襲及增殖能力的影響情況，探索CML66在腎癌細(xì)胞發(fā)生發(fā)展中可能的信號(hào)通路機(jī)制，為腎癌的治療提供可能的有效靶點(diǎn)，現(xiàn)報(bào)道如下。

1 材料與方法

1.1 材料

腎癌786-O細(xì)胞株購買于中科院上海公司;CML66、LIS1及Dynein抗體購買于Abcam公司;細(xì)胞增殖與毒性檢測試劑盒購買于同仁化學(xué)研究所;穿透小室購買于康寧公司;Matrigel膠購買于Becton， Dickinson and Company公司;脂質(zhì)體Lipofectamine 2000購買于Invitrogen公司。