摘要
目的:探讨黑素瘤分化相关基因-7/白细胞介素-24(mda-7/IL-24)对非小细胞肺癌细胞系H460增殖的影响。方法:将mda-7/IL-24插入真核表达载体pcDNA3中,构建重组表达载体pcDNA 3-mda-7/IL-24。按照DNA和脂质体(L ipofectam ine)的不同比例,将重组表达载体瞬时转染H460细胞,用RT-PCR检测mda-7/IL-24在不同转染条件下的表达,用MTT法检测转染重组表达载体后对H460细胞增殖的影响。结果:成功构建了mda-7/IL-24的重组表达载体,用SP6引物和mda-7/IL-24上游引物进行RT-PCR分析,在DNA和L ipofectam ine的比例为1μg∶2μl^1μg∶4μl时有目的基因表达。按照1μg∶2μl的比例瞬时转染重组表达质粒72 h后进行MTT分析,结果表明,与未转染细胞和转染空质粒细胞相比,重组质粒pcDNA3-mda-7/IL-24对肺癌细胞H460的增殖有明显抑制作用,抑制率为18.4%。结论:mda-7/IL-24对非小细胞肺癌细胞系H460增殖具有明显的抑制作用。
Objective：To analyze the role of human mda-7/IL-24 gene on the proliferation of human non-small cell lung cancer cell line （H460）. Methods ： The mda-7/IL-24 cDNA was subcloned into expression vector pcDNA3 and transient transfected in human lung carcinoma cell H460. The overexpressed mda-7/IL-24 was identified by RT-PCR using SP6 primer and mda-7/IL-24 upstream primer. The role of human mda-7/IL-24 on the proliferation of H460 cells was analyzed with MTT after transfection of pcDNA3-mda-7/IL-24 for 72 h. Results ：The mda-7/IL-24 cDNA was subcloned into pcDNA3 successfully. RT-PCR study showed that mda-7/IL-24 could express in H460 cells at different ratio of DNA and lipofectamine （1 μg ：2 μl- 1μg ： 4μl）. Compared with H460 cells and H460 cells transfected with pcDNA3, pcDNA3-mda -7/IL-24 could inhibit the proliferation of H460 cells. The inhibition rate was 18.4%. Conclusion：The mda-7/IL-24 can inhibit the proliferation of human non-small-cell lung carcinoma cell H460.
出处
《医学研究生学报》
CAS
2005年第9期779-783,共5页
Journal of Medical Postgraduates
