Wang Zhiyang, Liu Shuai, Ding Kejia, Ding Sentai, Li Chensheng, Lu Jiaju, Gao Dexuan, Zhang Tong, Bi Dongbin
Department of Urology, Shandong Provincial Hospital Affiliated to Shandong University, 324 Jingwu Street, Jinan, 250021, People's Republic of China.
Department of Digestive Diseases, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, 250021, People's Republic of China.
Tumour Biol. 2016 Nov;37(11):15133-15143. doi: 10.1007/s13277-016-5395-1. Epub 2016 Sep 27.
Renal cell carcinoma (RCC) accounts for 3 % of all adult malignancies and is the most lethal urological cancer. Livin is a member of the inhibitor of apoptosis protein (IAP) family, which is associated with tumor resistance to radiotherapy and chemotherapy. Clinical data also showed that patients with high tumor grades and stages have higher expression levels of Livin in RCC cells. Autophagy is a survival mechanism activated in response to nutrient deprivation. A possible role of Livin in the autophagy of RCC cells has not been investigated; therefore, this pioneer study was carried out. Livin was silenced in RCC cells (slow virus infection [SVI]-shLivin cells) by lentiviral transfection. Then, mRNA and protein expression levels in the transfected cells were assessed by quantitative fluorescence PCR and Western blotting, respectively. In addition, acridine orange staining and electron microscopy were used to assess autophagy in SVI-shLivin cells. The cisplatin IC50 values for RCC cells were measured by the CCK8 assay. Potent antitumor activities were observed in xenograft mouse models generated with Livin-silenced RCC cells in terms of delayed tumor onset and suppressed tumor growth. These results suggested that Livin silencing could increase the chemotherapeutic sensitivity of RCC cells to cisplatin and induce autophagic cell death. A possible mechanism of Bcl-2 and Akt pathway involvement was discussed specifically in this study. Overall, Livin silencing induces apoptotic and autophagic cell death and increases chemotherapeutic sensitivity of RCC cells to cisplatin.
肾细胞癌(RCC)占所有成人恶性肿瘤的3%,是最致命的泌尿系统癌症。Livin是凋亡抑制蛋白(IAP)家族的成员,与肿瘤对放疗和化疗的抗性有关。临床数据还显示,肿瘤分级和分期较高的患者肾癌细胞中Livin的表达水平更高。自噬是一种在营养剥夺时被激活的生存机制。Livin在肾癌细胞自噬中的可能作用尚未得到研究;因此,开展了这项开创性研究。通过慢病毒转染使肾癌细胞(慢病毒感染[SVI]-shLivin细胞)中的Livin沉默。然后,分别通过定量荧光PCR和蛋白质免疫印迹法评估转染细胞中的mRNA和蛋白质表达水平。此外,使用吖啶橙染色和电子显微镜评估SVI-shLivin细胞中的自噬。通过CCK8测定法测量肾癌细胞的顺铂IC50值。在用Livin沉默的肾癌细胞生成的异种移植小鼠模型中,在延迟肿瘤发生和抑制肿瘤生长方面观察到了强大的抗肿瘤活性。这些结果表明,Livin沉默可增加肾癌细胞对顺铂的化疗敏感性并诱导自噬性细胞死亡。本研究特别讨论了Bcl-2和Akt途径参与的可能机制。总体而言,Livin沉默诱导凋亡和自噬性细胞死亡,并增加肾癌细胞对顺铂的化疗敏感性。