摘要
目的通过miRNA介导RNAi沉默OPN表达探讨其对人肺腺癌细胞株A549顺铂(DDP)敏感性的影响。方法利用体外构建的靶向OPN的miRNA表达质粒瞬时转染A549,酶联免疫吸咐法(ELISA)检测转染后48h细胞OPN蛋白表达;转染后24、48、72、96h,四甲基偶氮唑蓝(MTT)法分别检测转染细胞和转染联合5μg/mL顺铂作用后细胞的增殖;于转染后24h联合不同浓度的顺铂作用48h,MTT法检测细胞增殖抑制率。结果(1)转染后48h,OPN蛋白表达水平下调了47.34%(P<0.01)。(2)转染后24h细胞增殖开始受抑制(P<0.05),48、72、96h细胞增殖被明显抑制,细胞增殖活性差异均有统计学意义(P<0.01)。(3)转染联合顺铂作用24h细胞增殖开始受抑(P<0.05),48、72、96h细胞增殖明显受抑,细胞增殖活性转染组与未转染组及非特异性转染组差异均有统计学意义(P<0.01)。转染后24h联合不同浓度的顺铂作用48h,当顺铂浓度为2.5、5、10、20μg/mL时,细胞增殖被明显抑制,细胞增殖抑制率差异均有统计学意义(P<0.01)。结论靶向OPN的miR-NA抑制人肺腺癌细胞A549增殖,增强细胞对DDP的敏感性。
Objective To explore the drug sensitivity of lung adenocarcinoma cell line A549 to cisplatin（DDP） by miRNA specific for OPN gene. Methods MicroRNA targeting OPN gene was transfected transiently to A549 cells by the expression plasmids which were constructed in vitro. The level of OPN protein in the A549 cells at 48 h after transfection was measured with ELISA. The cells incubated and non-incubated with 5μg/mL DDP were detected by MTT at 24,48,72 and 96 h after transfection. And the cells were cultured at different concentrations of cisplatin（DDP） for 48h,the growth inhibition rate was evaluated by MTT. Results （1）The OPN protein level was decreased by 47.34% at 48 h after transfection（P〈0.01 ）. （2）The cell proliferation started to be inhibited at 24 h after transfection（P〈0.05 ）, and was significantly inhibited at 48,72 and 96 h after transfection, with a significant difference（P〈0.01 ）. （3）After the treatment with 5μg/mL DDP, the cell proliferation started to be inhibited at 24h after transfection（P〈0.05）,and was significantly inhibited at 48,72 and 96h after transfection,with a significant difference in the cell proliferation activity between the transfection group and non-transfection group or non-specific transfeetion group（ P〈0.01 ）. At 2.5,5,10 and 20 μg/mL DDP,the cell proliferation was significantly inhibited, with a significant difference in the cell proliferation inhibition rate（P〈0.01）. Conclusion MicroRNA targeting OPN gene can effectively inhibite the cell proliferation of lung adenocarcinoma cell line and enhances the cell chemosensitization to DDP.
出处
《中国现代医生》
2010年第5期4-7,共4页
China Modern Doctor
基金
江西省自然科学基金资助(项目编号:200540036)
