通过RNA干扰下调hnRNP K可抑制人肺癌细胞的生长。
Downregulation of hnRNP K by RNAi inhibits growth of human lung carcinoma cells.
作者信息
Tang Fengming, Li Weimin, Chen Yan, Wang Dongmei, Han Juan, Liu Dan
机构信息
Department of Respiratory Medicine, West China Hospital of Sichuan University, Chengdu, Sichuan 610041, P.R. China ; Department of Respiratory Medicine, The First People's Hospital of Yibin, Yibin, Sichuan 644000, P.R. China.
Department of Respiratory Medicine, West China Hospital of Sichuan University, Chengdu, Sichuan 610041, P.R. China.
出版信息
Oncol Lett. 2014 Apr;7(4):1073-1077. doi: 10.3892/ol.2014.1832. Epub 2014 Jan 28.
Lung cancer is one of the most common malignancies worldwide, but its pathogenesis remains unknown. The current study examined the effects of heterogeneous nuclear ribonucleoprotein K (hnRNP K)-targeted small interfering RNA (siRNA) on the growth and apoptosis of lung cancer cells . The expression of hnRNP K was studied by the SP method of immunohistochemistry in lung tissue samples of 70 cases of lung cancer. hnRNP K siRNA were transfected into human lung cancer cell line, A549, using Lipofectamine 2000. Cells transfected with siRNAn and untreated served as controls. The inhibitory effect of siRNA on the expression of hnRNP K mRNA and protein was detected by reverse transcription polymerase chain reaction and western blot analysis. The change in cell cycling and cell apoptosis of siRNA-treated cells was detected by flow cytometry. The rates of positive hnRNP K expression in lung tumors of diameters ≤3, 3-5 and ≥5 cm, were 38.5, 95.2 and 91.7%, respectively. A significant difference was identified between lung tumors with diameters of ≤3 and ≥3 cm (P<0.01). The expression of hnRNP K mRNA was significantly inhibited in siRNA-transfected cells compared with that in control cells (P<0.05). Notably, hnRNP K protein decreased in hnRNP K siRNA-transfected cells, but exhibited no effect on the control groups. siRNA targeting human hnRNP K effectively inhibited the growth of lung cancer cell line, A549, and the distribution of the cell cycle. The apoptosis rate was 4.79% and the number of cells increased in the G0/G1 phase from 37.21 to 85.60% and decreased in the S and G2/M phases from 47.71 to 13.50% and 14.00 to 0.32%, respectively, following 24 h of transfection. hnRNP K siRNA promotes A549 apoptosis and the apoptosis rate was 4.79% (P<0.01). Therefore, hnRNP K siRNA may inhibit the proliferation of A549 cells. In addition, hnRNP K promotes the growth of lung cancer cells and, therefore, hnRNP K siRNA may inhibit the growth and increase the apoptosis of lung cancer cells.
肺癌是全球最常见的恶性肿瘤之一，但其发病机制尚不清楚。本研究检测了靶向异质性细胞核核糖核蛋白K（hnRNP K）的小干扰RNA（siRNA）对肺癌细胞生长和凋亡的影响。采用免疫组织化学SP法检测70例肺癌组织样本中hnRNP K的表达。使用Lipofectamine 2000将hnRNP K siRNA转染至人肺癌细胞系A549中。转染siRNAn的细胞和未处理的细胞作为对照。通过逆转录聚合酶链反应和蛋白质印迹分析检测siRNA对hnRNP K mRNA和蛋白表达的抑制作用。通过流式细胞术检测siRNA处理细胞的细胞周期变化和细胞凋亡情况。直径≤3 cm、3 - 5 cm和≥5 cm的肺肿瘤中hnRNP K阳性表达率分别为38.5%、95.2%和91.7%。直径≤3 cm和≥3 cm的肺肿瘤之间存在显著差异（P<0.01）。与对照细胞相比，转染siRNA的细胞中hnRNP K mRNA的表达受到显著抑制（P<0.05）。值得注意的是，hnRNP K siRNA转染的细胞中hnRNP K蛋白减少，但对对照组无影响。靶向人hnRNP K的siRNA有效抑制了肺癌细胞系A549的生长和细胞周期分布。转染24小时后，凋亡率为4.79%，G0/G1期细胞数量从37.21%增加到85.60%，S期和G2/M期细胞数量分别从47.71%减少到13.50%和从14.00%减少到0.32%。hnRNP K siRNA促进A549细胞凋亡，凋亡率为4.79%（P<0.01）。因此，hnRNP K siRNA可能抑制A549细胞的增殖。此外，hnRNP K促进肺癌细胞生长，因此，hnRNP K siRNA可能抑制肺癌细胞生长并增加其凋亡。
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