分泌肿瘤坏死因子的基因工程细胞对HepG2细胞的抑制作用
Inhibitory Effects on HepG2 Cells from Genetic Engineering Cells Secreted Tumor Necrosis Factor
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摘要: 目的 自行建立分泌人肿瘤坏死因子α的基因工程细胞，将其与肝癌细胞共培养,观察体外培养中分泌肿瘤坏死因子hTNF/293基因细胞表达的情况及对人肝癌细胞的抑制作用。方法(1)建立可稳定分泌人肿瘤坏死因子α/293细胞;采用RT-PCR、Western blot、ELISA和流式细胞仪等技术检测人肿瘤坏死因子表达和分泌;(2)观察体外培养中分泌肿瘤坏死因子hTNF/293基因细胞对人肝癌细胞的抑制作用,于不同的时间点,用MTT法检测490 nm下的吸光度值。结果RT-PCR、Western blot和ELISA等技术检测表明hTNFα/293 细胞组和TNFα阳性组对共培养的HepG2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TNF-α/293基因细胞可有效分泌hTNFα蛋白,并能分泌到细胞外;体外培养的人肿瘤坏死因子基因的工程细胞,所分泌肿瘤坏死因子α对人肝癌细胞增殖有明显抑制效应,且呈现出良好的数量依赖关系。Abstract: Objective To explore the effects of genetic engineering cells which secreted tumor necrosis factor(TNFα) co-cultured with human hepatic cancer cells HepG2 on the proliferation of cancer cells. Methods The expression and secreting of tumor necrosis factor was detected by RT-PCR,Western blot,ELISA and flow cytometry from the 293 cells which could stably secrete TNF-α.The inhibitory effects on the hepatic cancer cells HepG2 from the 293 cells (hTNFα/293) were detected by MTT assay under the wavenumber of 490 nm at different time. Results The results from RT-PCR,Western blot and ELISA indicated that hTNF-α /293 cells and TNF-α-positive group had significant inhibition on the proliferation of hepatic cancer cells in a dose-dependent manner. Conclusion The TNF-α/293 cells can effectively secrete TNF-α .And the TNF-α secreted from the genetic engineering cells has an inhibition on hepatic cancer cell proliferation in a dose-dependent manner.
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