shRNA靶向抑制CD74基因对人肾透明细胞癌细胞786-O生物学行为的影响

Inhibition of CD74 gene by shRNA affects the biological behaviors of human clear cell renal cell carcinoma cell line 786-O

目的探讨抑制人CD74基因后对肾透明细胞癌786-O细胞生长和侵袭能力的影响。方法通过慢病毒lentivirus-CD74转染肾透明细胞癌786-O细胞后,MTT法检测786-O细胞活力的变化,利用流式细胞学检测肿瘤细胞的凋亡变化情况,Transwell小室实验检测786-O细胞侵袭能力改变情况,采用Westernblot方法检测转染前后786-O细胞CD74及相关蛋白的变化情况。结果慢病毒转染786-O细胞后细胞中CD74蛋白水平表达明显低于对照组(P<0.05),抑制CD74蛋白表达水平后786-O细胞生长明显受到抑制,细胞活力降低,凋亡增加,细胞侵袭能力降低,且Survivin、MMP-2、MM-9蛋白也随之下降,其与对照组结果对比差异具有统计学差异(P<0.05)。结论利用慢病毒转染技术抑制CD74的表达可以抑制肾透明细胞癌786-O细胞的增值和侵袭能力,CD74基因可能作为癌基因在肾透明细胞癌中起作用。

Objective To explore the effect and mechanism of CD74 modulating growth and invasion on human clear cell renal cell carcinoma （ccRCC） cell line 786-0. Methods The 786-0 cells were transfected with lentivirus-CD74. The vitality of 786-O cells was detected with MTT assay. The apoptosis was detected with flow cytometry. The invasion was assessed with Transwell assay. The expression of CD74 was tested with Western blotting. Results After transfection, the expression of CD74 was decreased （P〈0.05）, the cell growth was significantly inhibited, apoptosis increased, invasion ability decreased, and the expression of Survivin, MMP-2 and MM-9 decreased （all P〈0.05）. Conclusions Knocking down of CD74 induced decreased cell proliferation and invasion, which may play a role in the oncogenes of ccRCC.