2,3-吲哚醌对MCF-7和HT-29细胞株的增殖抑制和凋亡诱导

Effectsof2, 3-Dioxoindolineon Anti-proliferation and Apoptosis n MCF-7 and HT-29 Cell Lines

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。

Objective： To investigate the effects of 2,3-dioxoindoline （ISA） on proliferation and apoptosis in human breast cancer cell（MCF-7） and human colon cancer cell（HT-29）. Methods： MTT assay was employed to examine the antiproliferative effect of ISA（50, 100, 200, 400 μmol·L^-1）for various time periods; the two cell lines treated with 400 μmol·L^-1 ISA for 48 h were stained with Hochest33258. Changes ofnucleolus were observed under fluorescent microscope. The expression levels of protein Bcl-2 and Bax were analyzed by Western Blotting. The levels ofbcl-2 mRNA and bax mRNA were examined by RT-PCR. Results：（1）ISA could inhibit the proliferation of the both cell lines obviously within a certain concentration. （2）Treated with 400 μmol· L^-1 ISA for 48 h, the typical apoptotic morphologic changes could be observed, including chromatin margination, nuclear condensation and apoptotic bodies. （3）Compared with that of the control, the expressions ofbcl-2 mRNA and protein were decreased significantly in a dose-dependent manner （P〈0.01 or P〈0.05）. However, bax mRNA and protein expression had no obvious change （P〉0.05）. Conclusions： ISA could inhibit the proliferation and further induce the apoptosis of the cell lines. Its mechanism may be related with the down-regulation of the expression of bcl-2.