跨膜蛋白106A在肺癌中的表达分析及其作用研究
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摘要
目的:探讨跨膜蛋白106A(transmembrane protein 106A,TMEM106A)在肺癌肿瘤中的表达及其与临床病理特征的关系,并进一步研究了该分子对肺癌细胞增殖的影响。方法:收集肺癌病人手术切除标本31例,提取组织RNA,采用荧光实时定量PCR实验,检测肺癌组织以及肺癌旁组织中TMEM106A mRNA的相对表达量。构建TMEM106A慢病毒表达载体Plenti-UBI/TMEM106A,转染至H1299细胞中,实现TMEM106A高表达,采用CCK-8细胞增殖实验和克隆形成实验观察TMEM106A过表达对肺癌细胞H1299增殖能力的抑制作用。结果:荧光实时定量PCR实验结果显示肺癌组织中,TMEM106A基因的mRNA相对表达量低于肺癌旁对照组。成功构建肺癌H1299-TMEM106A过表达细胞系,与H1299-control比较,细胞系H1299-TMEM106A中TMEM106A表达显著升高;增殖实验结果显示TMEM106A高表达显著抑制了肺癌细胞的增殖能力;克隆形成实验结果显示细胞系H1299-TMEM106A中高表达TMEM106A的肺癌细胞克隆形成能力明显低于H1299-control肺癌细胞(P<0.01)。结论:TMEM106A能抑制肺癌细胞的增殖。
AIM: To investigate the expression of transmembrane protein 106 A(TMEM106 A) in lung cancer and its relationship with clinicopathological features, and the effect of this molecule on the proliferation of lung cancer cells. METHODS: Thirty-one cases of lung cancer tissues were collected from patients with early stage lung cancer. Tissue RNA was extracted and real-time quantitative PCR was used to detect the relative expression of TMEM106 A mRNA in lung cancer tissues and para-cancer tissues. TMEM106 A lentiviral expression vector Plenti-UBI/TMEM106 A was constructed and transfected into H1299 cells to overexpress TMEM106 A. Cell proliferation assay and colony formation assay were used to observe the effect of TMEM106 A overexpression on the proliferation of lung cancer H1299 cells.RESULTS:Real-time quantitative PCR(qRT-PCR) showed that the relative expression of TMEM106 A gene mRNA in lung cancer tissues was lower than that in the control para-cancer tissues. H1299-TMEM106 A,the cell line overexpressed TMEM106 A, was successfully constructed. Compared with H1299-control, the expression of TMEM106 A in cell line H1299-TMEM106 A was significantly increased. The proliferation experiment showed that the high expression of TMEM106 A significantly inhibited the proliferation of lung cancer cells. The colony formation assay showed that the colony forming ability of lung cancer cells with high expression of TMEM106 A in cell line H1299-TMEM106 A was significantly lower than that of H1299-control lung cancer cells(P<0.01). CONCLUSION: TMEM106 A can inhibit the proliferation of lung cancer cells.
AIM: To investigate the expression of transmembrane protein 106 A(TMEM106 A) in lung cancer and its relationship with clinicopathological features, and the effect of this molecule on the proliferation of lung cancer cells. METHODS: Thirty-one cases of lung cancer tissues were collected from patients with early stage lung cancer. Tissue RNA was extracted and real-time quantitative PCR was used to detect the relative expression of TMEM106 A mRNA in lung cancer tissues and para-cancer tissues. TMEM106 A lentiviral expression vector Plenti-UBI/TMEM106 A was constructed and transfected into H1299 cells to overexpress TMEM106 A. Cell proliferation assay and colony formation assay were used to observe the effect of TMEM106 A overexpression on the proliferation of lung cancer H1299 cells.RESULTS:Real-time quantitative PCR(qRT-PCR) showed that the relative expression of TMEM106 A gene mRNA in lung cancer tissues was lower than that in the control para-cancer tissues. H1299-TMEM106 A,the cell line overexpressed TMEM106 A, was successfully constructed. Compared with H1299-control, the expression of TMEM106 A in cell line H1299-TMEM106 A was significantly increased. The proliferation experiment showed that the high expression of TMEM106 A significantly inhibited the proliferation of lung cancer cells. The colony formation assay showed that the colony forming ability of lung cancer cells with high expression of TMEM106 A in cell line H1299-TMEM106 A was significantly lower than that of H1299-control lung cancer cells(P<0.01). CONCLUSION: TMEM106 A can inhibit the proliferation of lung cancer cells.
引文
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[2] 陈万青,孙可欣,郑荣寿,等.2014年中国分地区恶性肿瘤发病和死亡分析[J].中国肿瘤,2018,23(1):1-14.
[3] 杨付红,张春娣.肺癌的研究现状[J].中国继续医学教育,2015,20(2):30-31.
[4] Cotter TG.Apoptosis and cancer:the genesis of a research field[J].Nat Rev Cancer,2009,9(7):501-507.
[5] Hanahan D,Weinberg R.Hallmarks of cancer:the next generation [J].Cell,2011,144(5):646-674.
[6] Wood HB.TMEM106B is a susceptibility locus for Ftld [J].Nat Rev Neurol,2010,6(4):184.
[7] Finch N,Carrasquillo MM,Baker M,et al.TMEM106B regulates progranulin levels and the penetrance of FTLD in GRN mutation carriers[J].Neurology,2011,76(5):467-474.
[8] Van dZJ,Van LT,Kleinberger G,et al.TMEM106B is associated with frontotemporal lobar degeneration in a clinically diagnosed patient cohort [J].Brain,2011,134(2):808-815.
[9] Xu D,Qu L,Hu J,et al.Transmembrane protein 106A is silenced by promoter region hypermethylation and suppresses gastric cancer growth by inducing apoptosis [J].J Cel Mol Med,2014,18(8):1655-1666.
[10] Wu CL,Xu J,Wang H,et al.TMEM106a is a novel tumor suppressor in human renal cancer [J].Kid Blood Press Res,2017,42(5):853.
[11] 许冬,姜静远,许晨彤,等.跨膜蛋白TMEM106A诱导肝癌细胞HepG2巨泡样死亡[J].中国生物化学与分子生物学报,2017,30(1):44-50.
[12] 钟良瑞,陈华,景晶,等.三叶青黄酮抑制肺癌A549细胞生长与蛋白酶体活性的关系研究[J].中国临床药理学与治疗学,2017,22(10):1123-1126.
[13] Elmore S.Apoptosis:a review of programmed cell death [J].Toxicol Pathol,2013,35:495-516.
[14] Horvitz HR.Genetic control of programmed cell death in the nematode Caenorhabditis elegans [J].Cell,1986,44(6):817-829.
[15] Sager R.Tumor suppressor genes in the cell cycle [J].Curr Opin Cell Biol,1992,4(2):155.
[16] Das PM,Singal R.DNA methylation and cancer [J].J Clin Oncol,2004,22(22):4632-4642.
[17] Davis CD,Uthus EO.DNA methylation,cancer susceptibility,and nutrient interactions [J].Exp Biol Med,2004,229(10):988.
[18] Brown R,Strathdee G.Epigenomics and epigenetic therapy of cancer [J].Trend Mol Med,2002,8(4):S43-S48.