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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 stably transfected hTNF-α / 293 cells on the proliferation of human colon carcinoma (LOVO) cells in vitro. Methods The mRNA and protein levels of hTNF-α / 293 and Hek-293 cells stably transfected were detected by RT-PCR and ELISA. In vitro, hTNF-α / 293 cells were co-cultured with LOVO cells. The proliferation of LOVO tumor cells was assayed by MTT assay at different time points. Results After transfection of Hek-293 cells with recombinant plasmid, hTNFα protein was detected by Western blot in vitro. The results showed that hTNFα / 293 cells and hTNFα-positive cells could significantly inhibit the proliferation of LOVO cells co-cultured with hTNFα and had a good inhibitory effect Effective relationship. The results suggest that stable transfection of hTNF-α / 293 significantly inhibited the proliferation of LOVO cells, and showed a good quantitative relationship. Because TNF-α / 293 gene cells can effectively hTNFα secretion of protein, and can be secreted to the outside of the cell.