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目的:探讨Testin基因(TES)在非小细胞肺癌(NSCLC)组织和细胞株中的表达及其对人肺癌A549细胞增殖、迁移、侵袭和凋亡的影响。方法:收集2015年1月至2015年12月在华中科技大学同济医学院附属同济医院手术切除的27例NSCLC患者的癌组织及癌旁组织标本,用Western blotting法检测癌组织和癌旁组织,以及正常人胚肺成纤维细胞株MRC5和肺癌细胞株A427、A549、H1299、LK2、PC9和SW900中TES蛋白的表达水平。应用短发卡RNA(sh RNA)瞬时转染肺癌细胞株A549干扰TES基因的表达,并进一步检测TES低表达对A549细胞增殖、迁移、侵袭以及凋亡的影响,同时检测凋亡相关蛋白Bax、Bcl-2和Caspase-3的表达。结果:在NSCLC组织和细胞株中TES蛋白的表达明显下降(均P<0.05)。sh TES干扰A549细胞后,TES m RNA和蛋白表达水平均显著下降(均P<0.05)。抑制TES表达显著增强A549细胞的增殖[(2.75±0.04)vs(1.79±0.06),P<0.05]、迁移[(52.3±2.6)%vs(19.7±1.4)%,P<0.05]和侵袭能力[(31.2±3.9)%vs(14.5±4.1)%,P<0.05],同时降低了细胞凋亡率[(8.2±1.1)%vs(23.1±1.7)%,P<0.05]。TES低表达使A549细胞Bax和Caspase-3蛋白表达明显下降(P<0.05)、Bcl-2蛋白表达明显升高(P<0.05)。结论:TES在NSCLC组织中呈低表达,TES表达下调具有促进肺癌细胞的增殖、迁移、侵袭并抑制凋亡等生物学效应,其有可能成为肺癌治疗一个新靶点。
Objective: To investigate the expression of Testin gene (TES) in non-small cell lung cancer (NSCLC) tissues and cell lines and its effect on proliferation, migration, invasion and apoptosis of human lung cancer A549 cells. Methods: Twenty-seven NSCLC patients with cancer resected from Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from January 2015 to December 2015 were collected for cancer tissue and paracancerous tissue. Western-blotting was used to detect the cancer tissue and paracancerous tissue, And normal human embryonic lung fibroblast cell line MRC5 and lung cancer cell lines A427, A549, H1299, LK2, PC9 and SW900 TES protein expression levels. Transient transfection of lung cancer cell line A549 with short hairpin RNA (shRNA) to interfere with the expression of TES gene and further test the effect of TES low expression on proliferation, migration, invasion and apoptosis of A549 cells. Meanwhile, apoptosis related proteins Bax, Bcl -2 and Caspase-3 expression. Results: The expression of TES protein in NSCLC tissues and cell lines was significantly decreased (all P <0.05). After TES interfered with A549 cells, TES mRNA and protein expression levels were significantly decreased (all P <0.05). Inhibition of TES expression significantly enhanced the proliferation of A549 cells [(2.75 ± 0.04 vs 1.79 ± 0.06, P <0.05], migration (52.3 ± 2.6% vs 19.7 ± 1.4%, P <0.05] [(31.2 ± 3.9)% vs (14.5 ± 4.1)%, P <0.05] and decreased the rate of apoptosis [(8.2 ± 1.1)% vs (23.1 ± 1.7)%, P <0.05]. The low expression of TES decreased the expression of Bax and Caspase-3 protein (P <0.05) and the expression of Bcl-2 protein in A549 cells (P <0.05). CONCLUSION: TES is low expressed in NSCLC tissues. The down-regulation of TES may promote the proliferation, migration, invasion and apoptosis of lung cancer cells. It may be a new therapeutic target for lung cancer.