靛玉红甲肟对HT-29细胞增殖和凋亡的影响及机制

Effect of indirubin-3’-monoxime on proliferation and apoptosis of human HT-29 cells

背景与目的:近年来有报道称靛玉红甲肟(indirubin-3’-monoxime)在体内外实验中对部分实体肿瘤细胞具有明显的抑制作用,但尚未见其对人结肠癌HT-29细胞影响的研究报道,因而本文旨在研究其对HT-29细胞增殖和凋亡的影响及其机制。方法:MTT法检测不同浓度靛玉红甲肟处理HT-29细胞后细胞的增殖活性,制作生长抑制曲线。流式细胞仪检测凋亡率,RT-PCR检测细胞凋亡抑制基因survivin和bcl-2及凋亡促进基因BaxmRNA的变化。结果:靛玉红甲肟明显的抑制HT-29的增殖,其作用表现为剂量依赖性和时间依赖性(F=11.25,P<0.01)。流式细胞仪检测发现:以10μmol/L的靛玉红甲肟处理HT-29细胞后,其凋亡率呈时间依赖性上升(F=195.25,P<0.01)。RT-PCR检测发现HT-29细胞survivin(F=78.75,P<0.01)和Bax(F=87.61,P<0.01)转录上升,而bcl-2转录显著下降(F=95.82,P<0.01)。结论:靛玉红甲肟对人结肠癌HT-29细胞具有明显的增殖抑制和诱导凋亡作用,其作用机制与下调bcl-2/Bax比例有关。

Background and purpose： In recent years indirubin-3-monoxime has been found to be capable of inhibiting some cell proliferation in vitro and in vivo studies, but human colon cancer HT-29 cells, therefore the purpose in this paper was to study the effect of indirubin-3-monoxime on proliferation and apoptosis of HT-29 cells and its associated mechanism. Methods： HT-29 cells were treated with indirubin-3-monoxime. The proliferative status of cells was measured by methabenzthiazuron （MTT） assay, flow cytometry （FCM） was used to measure the apoptosis rate. RT-PCR was used to measure the transcription of apoptosis suppressor gene bcl-2, survivin and apoptosis promoting gene Bax. Results： Indirubin-3-monoxime inhibited growth of HT-29 cells in a dose-dependent and timedependent manner （F=I 1.25, P〈0.01）. The apoptosis rate increased after the treatment by indirubin-3-monoxime at 10 ~mol/L. There were significant differences between different time groups （F=195.25, P〈0.01）. The transcription of survivin （F=78.75, P〈0.01） and Bax （F=87.61, P〈0.01） mRNA in HT-29 cells were increased; the transcription of bcl-2 was significantly decreased （F=95.82, P〈0.01）. Conclusion： Indirubin-3-monoxime has obviously inhibited proliferation and induce apoptosis of colon cancer HT-29 cells, its mechanism may be related to decrease the bcl-2/Bax ratio.