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目的:研究2,3-吲哚醌(2,3-dioxoindoline,ISA)对人乳腺癌细胞株MCF-7和人大肠癌细胞株HT-29的增殖与凋亡影响。方法:应用MTT比色法检测ISA对MCF-7和HT-29的生长抑制情况;Hochest33258荧光染色法检测细胞凋亡情况,Western Blotting法检测凋亡相关基因bcl-2和bax在蛋白水平上的表达;RT-PCR两步法检测bcl-2和bax在mRNA水平上的变化。结果:①ISA在一定浓度范围内能明显抑制细胞生长;②荧光染色可观察到细胞核染色质浓集,核浓缩核碎裂,并出现典型的凋亡小体;③bcl-2mRNA及蛋白的表达随药物浓度增大而逐渐减少,各实验处理组与阴性对照组比较存在统计学差异(P<0.05或P<0.01),baxmRNA及蛋白的表达随药物浓度增大而无明显变化,各组间比较不存在统计学差异(P>0.05)。结论:ISA能够抑制MCF-7和HT-29细胞增殖,并能进一步诱导其凋亡,其作用机制可能是与下调bcl-2表达有关。
AIM: To investigate the effects of 2,3-dioxoindoline (ISA) on the proliferation and apoptosis of human breast cancer cell line MCF-7 and human colorectal cancer cell line HT-29. Methods: The inhibitory effects of ISA on the growth of MCF-7 and HT-29 cells were detected by MTT colorimetric assay. The apoptosis of cells was detected by Hochest33258 staining. The protein level of bcl-2 and bax were detected by Western Blotting The expression of bcl-2 and bax at mRNA level was detected by RT-PCR. Results: ①ISA could significantly inhibit the cell growth in a certain range of concentration. ② The nuclear chromatin concentration was observed by fluorescence staining, nuclear fragmentation was observed and typical apoptotic bodies were found. ③ The expression of bcl-2 mRNA and protein was not significantly different with the drug (P <0.05 or P <0.01). The expression of bax mRNA and protein had no significant change with the increase of drug concentration, and the differences among the groups were not statistically significant There was a statistically significant difference (P> 0.05). Conclusion: ISA can inhibit the proliferation of MCF-7 and HT-29 cells and induce apoptosis of MCF-7 and HT-29 cells. Its mechanism may be related to the down-regulation of bcl-2 expression.