紫杉醇对CHMm细胞G_2/M期的阻滞及对周期相关因子的影响

Effects of paclitaxel on G_2/M phase arrest and cycle related factors in CHMm cells

为探讨紫杉醇对犬乳腺肿瘤细胞的周期阻滞及周期相关因子的表达影响,采用不同浓度紫杉醇以不同作用时间处理细胞,通过倒置显微镜、流式细胞仪(FCM)、SYBR染料法实时荧光定量PCR,研究细胞的形态学变化、G2/M期阻滞以及对抑癌基因p27、p53和原癌基因bcl-2表达的影响。结果显示,0.1μmol/L紫杉醇作用72h,犬乳腺肿瘤细胞(CHMm)生长状况极差,细胞大量凋亡。同时,0.1μmol/L紫杉醇作用48h,G2/M期阻滞明显,比例高达78.69%。紫杉醇对周期相关因子的影响,具有时间-剂量关系,p27、p53的表达均呈上升趋势,bcl-2的表达呈下降趋势。结果表明,紫杉醇能抑制细胞增殖,诱导细胞凋亡;促使细胞G2/M期阻滞和抑制细胞有丝分裂;并有促进p27、p53表达、抑制bcl-2表达的作用。

To explore the effects of paclitaxel on cell cycle arrest and changes in expression of cycle related factors,canine breast tumor cells（CHMm） were treated with paclitaxel in different concentration and for different time. The morphological changes,G2/M phase arrest and the expression changes of p27, p53 and bcl-2 were detected by inverted microscope,flow cytometry（FCM） and real time fluorescent quantitative PCR with SYBR. In result,CHMm cells grown badly and a large number of cells underwent apoptosis at concentration of 0. 1μmol/L for 72 h. G2/M phase arrest was significant at concentrations of 0. 1 μmol/L for 48 h, up to 78. 69 %. The effects of paclitaxel on cycle-related factors were in time-dose relationship,and p27 and p53 expressions showed an upward trend but bcl-2 was down regulated. The results showed that paclitaxel inhibited cell proliferation,induced apoptosis,promoted G2/M phase arrest and inhibited cell mitosis. Moreover, paclitaxel promoted the expression of tumor suppressor genes p27 and p53 but inhibited the expression of oncogene bcl-2.