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目的 研究天花粉蛋白(TCS)对胃癌多药耐药细胞细胞毒和诱导凋亡作用,探讨其治疗胃癌多药耐药的可能性。 方法 采用生长曲线、MTT、电镜、TUNEL染色和流式细胞仪等技术,研究TCS对胃癌多药耐药细胞SGC-7901/VCR和MKN-45/VCR的增殖抑制和诱导凋亡作用。 结果 TCS明显抑制胃癌多药耐药细胞SGC-7901/VCR和MKN-45/VCR的生长,其作用与对其本细胞的抑制作用相近。TCS对SGC-7901/VCR和MKN-45/VCR细胞有较强的细胞毒作用,TCS对SGC-7901和SGC-7901/VCR的IC50分别为28.6μg/mL和35.3μg/mL;对MKN-45和MKN-45/VCR的IC50分别为和10.67μg/mL和14.61μg/mL。TCS作用于MKN-45细胞后,光镜和电镜下能见到典型的细胞凋亡形态学特征:细胞核染色质致密浓缩,染色质断裂、凋亡小体形成等。TUNEL检测显示耐药细胞凋亡指数为2.8％～11.5％,流式细胞仪DNA直方图上出现典型的亚二倍体“凋亡峰”,耐药细胞凋亡率在4.73％～21.5％,细胞凋亡与TCS作用时间和浓度相关。 结论 TCS对胃癌多药耐药细胞有较强的细胞毒和诱导凋亡作用,具有治疗胃癌多药耐药的潜在价值。
Objective To study the cytotoxicity and apoptosis-inducing effect of trichosanthin (TCS) on gastric cancer multidrug resistant cells, and to explore the possibility of multidrug resistance in gastric cancer. Methods Growth inhibition, MTT, electron microscopy, TUNEL staining and flow cytometry were used to study the effects of TCS on the proliferation inhibition and apoptosis induction of gastric cancer multidrug resistant cells SGC-7901/VCR and MKN-45/VCR. Results TCS significantly inhibited the growth of gastric cancer multidrug resistant cells SGC-7901/VCR and MKN-45/VCR, and its effect was similar to that of their own cells. TCS had a strong cytotoxic effect on SGC-7901/VCR and MKN-45/VCR cells. The IC50 of TCS on SGC-7901 and SGC-7901/VCR was 28.6 μg/mL and 35.3 μg/mL, respectively; on MKN- The IC50s of 45 and MKN-45/VCR were 10.67 μg/mL and 14.61 μg/mL, respectively. After TCS acts on MKN-45 cells, the typical morphology of apoptosis can be seen under light and electron microscopes: dense nuclear chromatin, chromatin breakdown, and apoptotic body formation. The TUNEL assay showed that the apoptotic index of drug-resistant cells ranged from 2.8% to 11.5%, and typical subdiploid “apoptotic peaks” appeared on the flow cytometry DNA histogram. The apoptotic rate of drug-resistant cells ranged from 4.73% to 21.5%. Apoptosis is related to the time and concentration of TCS action. Conclusion TCS has a strong cytotoxicity and apoptosis-inducing effect on multidrug resistant gastric cancer cells, and it has potential value in the treatment of gastric cancer multidrug resistance.