Zhao Xiaozhen, Xu Zhenye, Wang Zhongqi, Wu Zhonghua, Gong Yabin, Zhou Lijuan, Xiang Yi
Department of Oncology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, P.R. China.
Experiment Center for Science and Technology, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, P.R. China.
Mol Med Rep. 2015 Jul;12(1):960-6. doi: 10.3892/mmr.2015.3471. Epub 2015 Mar 11.
The expression of integrin-linked kinase (ILK) has been reported to be involved in the regulation of integrin-mediated processes, including cancer cell proliferation, migration and invasion. Previous studies have demonstrated that inhibition of ILK may be an underlying approach for treating cancer. However, whether the knock down of ILK affects growth and apoptosis of lung cancer cells remains to be elucidated. Importantly, whether downregulation of ILK increases the sensitivity of lung cancer cells to cisplatin and amplifies cell apoptosis also remains to be elucidated. In the present study, ILK downregulation was mediated by lentivirus-mediated RNA interference. The expression levels of associated genes were determined by reverse-transcription quantitative polymerase chain reaction and western blotting. Cell proliferation was evaluated using a modified 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and clone formation assay. The cell cycle and apoptosis were analyzed using flow cytometry. The current data revealed that lentivirus-mediated ILK gene silencing alone inhibited A549 cell proliferation and promotes cell cycle arrest, however, had no detectable effect on cell apoptosis. However, combined treatment with lentivirus-mediated ILK interference and cisplatin chemotherapy induced significantly more cell apoptosis than mono-chemotherapy or knockdown. The increased cell apoptosis and proliferation inhibition were attributed to abnormal downstream protein expression of ILK, including phospho-glycogen synthase kinase 3β, p-AKT, activator protein-1, β-catenin, cyclin D1 and matrix metalloproteinase-9. ILK inhibition may suppress the proliferation of A549 and increase A549 sensitivity to cisplatin. The combined treatment of ILK gene knockdown and chemotherapy has the potential to improve anticancer efficacy.
据报道，整合素连接激酶（ILK）的表达参与整合素介导的过程调控，包括癌细胞的增殖、迁移和侵袭。先前的研究表明，抑制ILK可能是治疗癌症的一种潜在方法。然而，敲低ILK是否会影响肺癌细胞的生长和凋亡仍有待阐明。重要的是，ILK的下调是否会增加肺癌细胞对顺铂的敏感性并放大细胞凋亡也仍有待阐明。在本研究中，通过慢病毒介导的RNA干扰下调ILK。通过逆转录定量聚合酶链反应和蛋白质印迹法测定相关基因的表达水平。使用改良的3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐试验和克隆形成试验评估细胞增殖。使用流式细胞术分析细胞周期和凋亡。目前的数据显示，单独的慢病毒介导的ILK基因沉默可抑制A549细胞增殖并促进细胞周期停滞，但对细胞凋亡没有可检测到的影响。然而，慢病毒介导的ILK干扰和顺铂化疗联合治疗诱导的细胞凋亡明显多于单一化疗或基因敲低。细胞凋亡增加和增殖抑制归因于ILK下游蛋白表达异常，包括磷酸化糖原合酶激酶3β、p-AKT、激活蛋白-1、β-连环蛋白、细胞周期蛋白D1和基质金属蛋白酶-9。抑制ILK可能会抑制A549的增殖并增加A549对顺铂的敏感性。ILK基因敲低与化疗联合治疗有可能提高抗癌疗效。
