摘要
目的通过体外实验研究观察稳定转染hTNF-α/293基因细胞对人结肠癌(LOVO)细胞生长增殖影响。方法采用RT-PCR和ELISA检测稳定转染hTNF-α/293基因细胞和Hek-293细胞mRNA和蛋白表达水平。观察体外培养中,hTNF-α/293基因细胞与LOVO细胞共同培养,于不同的时间点,采用噻唑蓝MTT法检测LOVO肿瘤细胞增殖的活性。结果重组质粒转染Hek-293细胞后,Western blot检测到了hTNFα蛋白体外表达,检测表明hTNFα/293细胞组和hTNFα阳性组对共培养的LOVO细胞的增殖具有明显的抑制作用,且具有良好的量效关系。结论提示稳定转染hTNF-α/293对LOVO细胞增殖有明显抑制效应,且呈现出良好的数量依赖关系。由于TNF-α/293基因细胞可有效分泌hTNFα蛋白,并能分泌到细胞外。
objective To observe the effects of co-cuhure of hTNFα-sercreting and human colon cancer cells LOVO on the proliferation of cancer cells. Methods The stable transfected hTNF-α/293 , mRNA of Hek-293 cell and protein expression were detected by RT-PCR and ELISA. The positive group was added hTNF-α factor,and MTT assay was applied under the optical density 490 nm. Through human tumor cell proliferation inhibition experiment,the inhibitory effects on colon cancer cells （LOVO） proliferation were observed. Results hTNF-α/ 293 cells and hTNF-α-positive group showed a significant lower A, which suggested that hTNF-α/293 cells and hTNF-α-positive group had significant inhibition on the proliferation of colon cancer cells. Conclusion The inhibition of hTNF-α secreted by hTNF-α/293 cells on co-lon cancer cell proliferation shows significant dose-effect dependency, and hTNF-α expresses a considerable inhibition on the colon cancer cell proliferation as positive drug.
出处
《局解手术学杂志》
2013年第6期591-593,共3页
Journal of Regional Anatomy and Operative Surgery
基金
十二五国家技术研究发展计划(863计划)基金项目(2012AA020808)(2012-2015)
