Liu Dabei, Mei Xingke, Wang Linlin, Yang Xueying
Department of Thoracic Surgery, The Fourth Affiliated Hospital of China Medical University, Shenyang, Liaoning 110032, P.R. China.
Mol Med Rep. 2017 Jun;15(6):3963-3968. doi: 10.3892/mmr.2017.6545. Epub 2017 May 3.
The Rho kinase pathway has previously been reported to possess a close relationship with the growth, migration and invasion of lung cancer cells. However, the molecular mechanisms underlying the effects of this pathway on lung cancer cells are still elusive. The aim of the present study was to investigate the effects and underlying molecular mechanisms of Ras homolog family member A (RhoA) on the proliferation and apoptosis of SPCA1 lung carcinoma cells. Stable SPCA1 lung cancer cell lines, in which RhoA expression was silenced by small interfering RNA, were isolated following Geneticin screening. Inhibition of RhoA expression significantly decreased the proliferation of SPCA1 lung cancer cells, whereas apoptosis was significantly increased (P<0.01) as determined by the MTS tetrazolium assay and flow cytometry analysis, respectively. At the molecular level, knockdown of RhoA resulted in the significant activation of caspase‑3 (P<0.01), and a significant reduction in the levels of phosphorylated signal transducer and activator of transcription (phospho‑STAT3; P<0.01), as determined by western blotting. The results suggested that RhoA knockdown prevents cell proliferation and induces apoptosis in SPCA1 lung cancer cells. Furthermore, the underlying mechanisms responsible for these effects may include the activation of caspase‑3 and the reduction of phospho‑STAT3 levels.
此前有报道称，Rho激酶通路与肺癌细胞的生长、迁移和侵袭密切相关。然而，该通路对肺癌细胞产生影响的分子机制仍不清楚。本研究的目的是探讨Ras同源家族成员A（RhoA）对SPCA1肺癌细胞增殖和凋亡的影响及其潜在分子机制。通过遗传霉素筛选，分离出RhoA表达被小干扰RNA沉默的稳定SPCA1肺癌细胞系。分别通过MTS四唑盐检测和流式细胞术分析确定RhoA表达的抑制显著降低了SPCA1肺癌细胞的增殖，而凋亡则显著增加（P<0.01）。在分子水平上，通过蛋白质印迹法确定，RhoA的敲低导致半胱天冬酶-3显著激活（P<0.01），磷酸化信号转导子和转录激活子（磷酸化STAT3）水平显著降低（P<0.01）。结果表明，RhoA敲低可阻止SPCA1肺癌细胞的增殖并诱导其凋亡。此外，造成这些影响的潜在机制可能包括半胱天冬酶-3的激活和磷酸化STAT3水平的降低。
