重组改构TNF-α与顺铂、吉西他滨联合对H1299细胞系杀伤作用

The killing effect of recombinant mutant human tumor necrosis factor combined with cisplatin or gemcitabine on H1299 cell line

目的观察重组改构肿瘤坏死因子(TNF-α)在体外和顺铂、吉西他滨联合对人肺腺癌H1299细胞系杀伤作用。方法MTT法检测不同浓度TNF-α、顺铂、吉西他滨单药对H1299细胞的抑制率,一定浓度的TNF-α与不同浓度顺铂或吉西他滨联合时对H1299细胞的抑制率,有或无TNF-α作用下H1299细胞的平板克隆形成率和细胞生长曲线,流式细胞仪检测各组细胞凋亡率。结果在不同浓度TNF-α作用下,H1299细胞抑制率均为负值,与药物浓度无明显相关性;TNF-α对H1299细胞的细胞生长曲线和克隆形成率均无明显影响;TNF-α与顺铂联合时在顺铂浓度为10μg/m l时H1299细胞抑制率,明显高于单纯顺铂的作用(P<0.05);TNF-α与吉西他滨联合时在吉西他滨浓度为20、40μg/m l时对H1299细胞抑制率明显高于单纯吉西他滨的作用(P<0.05)。流式细胞分析各组细胞凋亡率与MTT结果一致。结论在体外重组改构肿瘤坏死因子与顺铂或吉西他滨联合应用明显增强后者对H1299细胞的杀伤作用,具有化疗增敏的作用。

Objective To study the killing effect of combining recombinant mutant human tumor necrosis factor （TNF-α） with cisplatin or gemcitabine on H1299 cell line in vitro. Methods MTT method was applied to detect the inhibition ratio of H1299 cells treated by different concentrations of TNF-α, cisplatin or gemcitabine alone, anti 8000 U/ml TNF-α combined with different concentrations of cisplatin or gemcitabine. Cell growth curve and cloning efficiency of H1299 cells treated by TNF-α or nothing were observed. The apoptotic rate of deferent groups were detected by tlow cytometer. Results The inhibition ratio were all minus after H1299 cells treated by different concentration of TNF-α. The cell growth curves and cloning efficiency of H1299 cells treated by TNF-α or nothing showed no evident difference. The inhibition ratio of TNF-α combined with 10μg/ml cisplatin or 20 μg/ml, 40μg/ml gemcitabine were remarkably higher than that of the later in the same concentration alone（P〈0.05）. The flow cytometry data of cell apoptosis showed the same result as MTT method＇s. Conclusion Our data suggest that TNF-α has no proliferative or inhibitive effect on H1299 cell line. But when combined with TNF-α, the killing effect of cisplatin or gemcitabine on H1299 cells was markedly enhanced.