Yu Nengwang, Fu Shuai, Liu Yubao, Xu Zhonghua, Liu Yi, Hao Junwen, Wang Baocheng, Zhang Aimin
Urology Department, General Hospital of Jinan Military Command, 25 Shifan Road, Jinan, Shandong, 250031, China.
Shandong Cancer Hospital & Institute, Shandong Academy of Medical Sciences, 440 Jiyan Road, Jinan, 250117, China.
J Exp Clin Cancer Res. 2015 Sep 29;34:107. doi: 10.1186/s13046-015-0224-8.
The present study examined the role of microRNA (miR)-96 in renal cell carcinoma (RCC) invasion.
The expression of miR-96 was detected by quantitative reverse transcription-polymerase chain reaction in human RCC cell lines with high (Caki-1) and low (786-O) metastatic potential. Invasive ability and Ezrin expression were assessed in Caki-1 and 786-O cells transfected with a miR-96 mimic or inhibitor using wound healing assays, Transwell assays and western blotting. Expression of miR-96 and Ezrin was also examined in primary RCC samples from 17 patients with metastatic disease and 46 patients who maintained remission during a follow-up period of 37 months.
miR-96 expression was significantly lower in Caki-1compared to786-O cells. The invasive ability of Caki-1 and 786-O cells increased following transfection of cells with miR-96 inhibitor, whereas it decreased following transfection with miR-96 mimic. Ezrin levels were negatively correlated with miR-96 in RCC, and inhibition of Ezrin expression suppressed the miR-96-induced change in invasive ability. The negative correlation between miR-96 and metastasis/Ezrin expression was also observed in human RCC specimens.
These results suggest that miR-96 suppresses RCC invasion by modulating Ezrin expression.
本研究检测了微小RNA(miR)-96在肾细胞癌(RCC)侵袭中的作用。
采用定量逆转录-聚合酶链反应检测具有高转移潜能(Caki-1)和低转移潜能(786-O)的人肾癌细胞系中miR-96的表达。使用伤口愈合试验、Transwell试验和蛋白质印迹法评估转染了miR-96模拟物或抑制剂的Caki-1和786-O细胞的侵袭能力和埃兹蛋白表达。还检测了17例转移性疾病患者和46例在37个月随访期内保持缓解的患者的原发性肾癌细胞样本中miR-96和埃兹蛋白的表达。
与786-O细胞相比,Caki-1细胞中miR-96的表达显著降低。用miR-96抑制剂转染细胞后,Caki-1和786-O细胞的侵袭能力增强,而用miR-96模拟物转染后,侵袭能力降低。在肾癌细胞中,埃兹蛋白水平与miR-96呈负相关,抑制埃兹蛋白表达可抑制miR-96诱导的侵袭能力变化。在人肾癌细胞标本中也观察到miR-96与转移/埃兹蛋白表达之间的负相关。
这些结果表明,miR-96通过调节埃兹蛋白表达来抑制肾癌细胞的侵袭。