Tang Feng-ming, Li Wei-min, Chen Yan, Wang Dong-mei, Han Juan
Department of Respiratory Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2008 Sep;39(5):823-6.
To examine the effects of hnRNP K on the growth and apoptosis of lung adencarcinoma cells in vitro.
The expressions of hnRNP K were detected by the SP method of immunohistochemistry in 70 lung adencarcinoma tissue samples. The hnRNP K siRNA was transfected into human lung adencarcinoma cell line A549 with Lipofectamine 2000. The 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 RT-PCR and Western blot. The change in cell cycling and cell apoptosis of siRNA-treated cells was detected by flow cytometry.
Positive expressions of hnRNP K were found in 38.5%, 95.2% and 91.7% of lung adencarcinoma tissues in tumors with a diameter of < or =3 cm, 3-5 cm and > or =5 cm respectively (P < 0.01). The expressions of hnRNP K mRNA were significantly inhibited in siRNA- transfected cells compared with those in controls (P < 0.05). hnRNP K protein decreased in hnRNP K siRNA-transfected cells. siRNA targeting human hnRNP K inhibited the growth of lung adencarcinoma cell line A549 and the distribution of the cell cycle, with an apoptosis rate of 4.79%. Twenty four hours after transfection, the number of cells in G0/G1 phase increased from 37.21% to 85.60%, while the number of cells in S and G2/M phases decreased from 47.71% and 13.00% to 13.50% and 0.32%, respectively.
The expression of hnRNP K is closely correlated with the size of lung adencarcinoma. hnRNP K siRNA inhibits the growth aNd increases the apoptosis of lung adencarcinoma cells.
探讨异质性核糖核蛋白K（hnRNP K）对肺腺癌细胞体外生长及凋亡的影响。
采用免疫组织化学SP法检测70例肺腺癌组织标本中hnRNP K的表达。用脂质体2000将hnRNP K小干扰RNA（siRNA）转染人肺腺癌细胞系A549。转染siRNA的细胞和未处理的细胞作为对照。采用逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测siRNA对hnRNP K mRNA和蛋白表达的抑制作用。用流式细胞术检测经siRNA处理的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的肺腺癌组织中hnRNP K的阳性表达率分别为38.5%、95.2%和91.7%（P<0.01）。与对照组相比，转染siRNA的细胞中hnRNP K mRNA的表达明显受到抑制（P<0.05）。hnRNP K siRNA转染细胞中hnRNP K蛋白减少。靶向人hnRNP K的siRNA抑制肺腺癌细胞系A549的生长和细胞周期分布，凋亡率为4.79%。转染24小时后，G0/G1期细胞数量从37.21%增加到85.60%，而S期和G2/M期细胞数量分别从47.71%和13.00%减少到13.50%和0.32%。
hnRNP K的表达与肺腺癌大小密切相关。hnRNP K siRNA抑制肺腺癌细胞生长并增加其凋亡。
