Zhuang Li, Shen Li-Da, Li Kun, Yang Run-Xiang, Zhang Qin-Yong, Chen Yun, Gao Chun-Lin, Dong Chao, Bi Qing, Tao Jing-Nan, Wang Xiao-Nan, Tian Qing
Department of Medical Oncology, Yunnan Cancer Hospital, Kunming Medical University, Kunming, Yunnan 650118, P.R. China.
Department of Cardiology, The First People's Hospital of Kunming, Yunnan 650011, P.R. China.
Mol Med Rep. 2015 Jul;12(1):547-52. doi: 10.3892/mmr.2015.3372. Epub 2015 Feb 18.
Livin is a novel member of the inhibitor of apoptosis protein family that has been reported to be overexpressed in various types of human malignancy. Although several studies have demonstrated that livin may be used as an effective target for tumor therapy, few studies have investigated its role in human lung adenocarcinoma. In the present study, two different methods were used in order to investigate the tumor‑suppressing effect of livin in human lung adenocarcinoma cells. Firstly, small interfering (si)RNA technology was used to down regulate livin expression; siRNA-mediated knockdown of livin was confirmed using reverse transcription quantitative polymerase chain reaction and western blot analysis, and cell proliferations was assessed using an MTT assay in vitro. Secondly, inhibition of livin expression was induced through the synergistic inhibitory effect between flavopiridol and tumor necrosis factor‑related apoptosis-inducing ligand (TRAIL). Experimental results revealed that, following transfection of the livin gene-silencing vector, the gene expression of livin was markedly decreased, SPC-A1 cell proliferation was significantly reduced and the therapeutic effect of the chemotherapy drug cisplatin was markedly improved. This growth inhibitory effect was also observed in the flavopiridol and TRAIL combination treatment group. In the flavopiridol and TRAIL combination treatment group, the protein expression of livin was significantly reduced and the survival rate of SPC‑A1 cells was significantly lower than the flavopiridol and TRAIL single operation group. In conclusion, the RNA silencing and the synergistic inhibitory effect between flavopiridol with TRAIL was able to effectively inhibit the expression of livin, significantly decrease SPC-A1 tumor cell proliferation and significantly enhance sensitivity to the chemotherapy drug cisplatin. These findings suggest that livin may be used as a novel target for tumor gene therapy.
Livin是凋亡抑制蛋白家族的一个新成员,据报道在各种类型的人类恶性肿瘤中过表达。尽管多项研究表明Livin可能作为肿瘤治疗的有效靶点,但很少有研究探讨其在人肺腺癌中的作用。在本研究中,采用两种不同方法来研究Livin在人肺腺癌细胞中的抑癌作用。首先,使用小干扰(si)RNA技术下调Livin表达;通过逆转录定量聚合酶链反应和蛋白质印迹分析证实siRNA介导的Livin敲低,并在体外使用MTT法评估细胞增殖。其次,通过黄酮哌啶醇与肿瘤坏死因子相关凋亡诱导配体(TRAIL)之间的协同抑制作用诱导Livin表达的抑制。实验结果显示,转染Livin基因沉默载体后,Livin的基因表达明显降低,SPC-A1细胞增殖显著减少,化疗药物顺铂的治疗效果明显改善。在黄酮哌啶醇和TRAIL联合治疗组中也观察到这种生长抑制作用。在黄酮哌啶醇和TRAIL联合治疗组中,Livin的蛋白表达显著降低,SPC-A1细胞的存活率明显低于黄酮哌啶醇和TRAIL单药治疗组。总之,RNA沉默以及黄酮哌啶醇与TRAIL之间的协同抑制作用能够有效抑制Livin表达,显著降低SPC-A1肿瘤细胞增殖,并显著增强对化疗药物顺铂的敏感性。这些发现表明Livin可能作为肿瘤基因治疗的新靶点。