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hnRNP K在肺腺癌中表达及其作用的研究 被引量:2

Expression of hnRNP K in Lung Adenocarcinoma Cells
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摘要 目的研究核内不均一核糖核蛋白K(hnRNP K)在肺腺癌组织中的表达及其对肺腺癌细胞生长及凋亡的影响。方法采用SP免疫组织化学方法对经手术切除病理确诊为肺腺癌的70例肺癌组织标本进行hnRNP K蛋白表达的检测,根据不同肿瘤的直径,计算阳性表达率;构建hnRNP K siRNA表达载体,采用Lipofectamine 2000瞬时转染A549细胞24h后,RT-PCR、Western blot检测A549细胞hnRNP K mRNA及其蛋白表达量;以流式细胞仪检测重组质粒hnRNP K siRNA及siRNAn转染A549细胞后细胞周期及凋亡的变化。结果肺腺癌组织中肿瘤直径≤3cm、3~5cm、≥5cm的hnRNP K的阳性表达率分别为38.5%、95.2%、91.7%;hnRNP K siRNA转染24h后的A549细胞hnRNPK mRNA及蛋白表达明显被抑制(P〈0.01);hnRNP K siRNA能显著抑制A549的生长及细胞周期的分布,G0/G1的细胞数从37.21%增加到85.60%,S和G2/M的细胞数分别从47.71%、13.00%减少到13.50%和0.32%,差异均有统计学意义(P〈0.01)。hnRNP K siRNA能促进A549的凋亡,其凋亡率达到4.79%(P〈0.01)。结论hnRNP K能促进肺腺癌细胞的生长;hnRNP K siRNA可抑制肺腺癌细胞的生长,促进其凋亡。 Objective To examine the effects of hnRNP K on the growth and apoptosis of lung adencarcinoma cells in vitro. Methods 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 eytometry. Results 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 ≤3 cm, 3-5 cm and ≥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. Conclusion 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.
出处 《四川大学学报(医学版)》 CAS CSCD 北大核心 2008年第5期823-826,共4页 Journal of Sichuan University(Medical Sciences)
基金 教育部博士点基金(批准号20050610056)资助
关键词 肺腺癌 核内不均一核糖核蛋白K SIRNA Lung adencarcinoma Heterogeneous nuclear ribonucleoprotein K siRNA
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参考文献10

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