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目的 :研究重组人肿瘤坏死因子α(rh TNF-α)和 5 - Fu联合诱导结肠癌细胞凋亡及其对 bcl- 2基因表达的影响。方法 :应用 MTT法检测 TNF-α、5 - Fu对结肠癌细胞 SW- 4 80的细胞毒作用 ;应用末端标记法 (TU NEL法 )研究结肠癌 SW- 4 80细胞凋亡 ;应用免疫组化法研究 bcl- 2基因的表达。结果 :3× 10 6 U /L的 rh TNF-α联合不同剂量的 5 - Fu作用于 SW- 4 80细胞 4 8h后对 SW- 4 80细胞生长具有协同抑制作用 ;联合应用 3× 10 6 U/L的 TNF-α,12 5 mg/L的 5 - Fu作用于 SW- 4 80细胞 4 8h后 ,TU NEL法显示染色阳性细胞比对照组显著增加 (P<0 .0 1) ;免疫组化结果显示 ,联合用药组 bcl- 2基因表达阳性的细胞百分率较对照组显著减低 (P<0 .0 1)。结论 :联合应用 TNF-α、5 - Fu有协同诱导 SW- 4 80细胞凋亡及抑制 bcl- 2基因的表达的作用。
Objective: To investigate the effect of recombinant human tumor necrosis factor-alpha (rh TNF-alpha) and 5-FU on apoptosis of colon cancer cells and its effect on bcl-2 gene expression. METHODS: MTT assay was used to detect the cytotoxicity of TNF-α and 5-FU on colon cancer cell line SW-480 and TUNEL was used to study the apoptosis of colon cancer SW-480 cells. Method to study the expression of bcl-2 gene. RESULTS: 3 × 10 6 U/L rhTNF-α combined with different doses of 5-FU in SW-480 cells had a synergistic inhibitory effect on the growth of SW-480 cells after 48 hours; combined application of 3 × 10 6 U TU-NEL assay showed that the positive cells were significantly increased compared with the control group (P < 0.01), and the immunization group was treated with TNF-α of /L and TNF-α of 12 5 mg/L for 48 hours. The results showed that the percentage of cells positive for bcl-2 expression in the combination group was significantly lower than that in the control group (P < 0.01). Conclusion : The combined use of TNF-α and 5-FU has a synergistic effect on apoptosis of SW-480 cells and inhibition of bcl-2 gene expression.