摘要
背景与目的目前已知环氧化酶2(COX2)在非小细胞肺癌(NSCLC)的浸润、发展及转移中有重要意义,抑制COX2的表达可能抑制NSCLC的发生及发展。用反义COX2重组载体转染高表达COX2的人非小细胞肺癌细胞株H1299,观察其对H1299细胞的生长抑制作用及顺铂敏感性的影响。方法通过脂质体介导将COX2反义核酸质粒转入H1299细胞,经氨基糖苷磷酸转移酶(Geneticin,G418)筛选获得稳定表达COX2反义核酸的NSCLC转染细胞系;采用逆转录聚合酶链式反应(RTPCR)技术检测COX2mRNA的表达水平;应用蛋白质印迹分析(WesternBlot)技术检测COX2蛋白表达水平的变化;应用四甲基偶氮唑蓝(MTT)比色法检测COX2反义核酸对H1299细胞增生的影响及其对H1299细胞的顺铂敏感性的影响。结果反义COX2基因转染后的H1299细胞中COX2mRNA表达水平明显下调,COX2蛋白表达水平明显减低。转染后的H1299细胞的生长速率明显下降(P<0.05),对顺铂的半数存活浓度(IC50)值由转染前的3.8mg/l降低为1.8mg/l(P<0.05)。结论COX2反义核酸导入不仅可抑制H1299细胞的增生,而且可增加H1299细胞对顺铂的敏感性。
Background and objective At present, it has been known that cyclooxygenase-2 （COX-2） plays a crucial role in invasion, development and metastasis of non-small cell lung cancer （NSCLC）. In order to explore whether the expression of COX-2 inhibits the occurrence and development of NSCLC, antisense vector of human COX-2 gene is transfected into COX-2 highly expressing NSCLC cell line H1299 and its effects on proliferation and sensitivity to cisplatin of H1299 are analysed. Methods H1299 cells were transfected with antisense vector of human COX-2 gene using LipoVecTM transfecting technique. Transfected cells were selected with Geneticin （G418）. The COX-2 mRNA level was examined by using reverse polymerase chain reaction （RT-PCR）. The COX-2 protein level was examined by Western Blot. The proliferative status and sensitivity to cisplatin of cells was measured by methabenzthiazuron （MTT） assay. Results RT-PCR showed a lower COX2 mRNA level in transfected cells. The level of COX-2 protein was decreased apparently. The proliferative index of the transfected cells decreased significantly （P〈0.05）. The IC50 value of cisplatin decreased remarkably in transfected cells （1.8 mg/l） compared with that in H1299 cells without transfection （3.8 mg/l）（P〈0.05）. Conclusion Transfection with antisense vector of human COX-2 gene can not only inhibit the proliferation of H1299 cells, but also increase the sensitivity to cisplatin of H1299 cells.
出处
《中国肺癌杂志》
CAS
2005年第5期424-427,共4页
Chinese Journal of Lung Cancer
