去甲斑蝥素诱导肿瘤干细胞凋亡作用研究

Induction of apoptosis of cancer stem cells by norcantharidin

目的探究去甲斑蝥素(NCTD)对肿瘤干细胞促凋亡作用。方法分别以UMUC-3膀胱癌和Hep G2肝癌细胞为研究对象,通过无血清、低吸附、单克隆的方法筛选肿瘤干细胞(CSC),实时荧光定量PCR(q RT-PCR)检测CSC干性标志物表达变化,MTS法检测NCTD及紫杉醇(PTX)对CSC的抑制作用,流式细胞术检测细胞凋亡。结果 2种细胞经筛选获得成球生长细胞克隆,其干性基因NANOG、OCT4、ABCG2、CD133、CD34表达明显升高,释放入含血清培养基后干性基因表达下调。经筛选所得的CSC对PTX高度耐药,但对NCTD耐药性低。NCTD可诱导CSC凋亡,但对干性基因表达没有显著影响。结论 NCTD诱导凋亡杀伤CSC,但对CSC分化没有显著影响。

Objective To investigate the effects of norcantharidin（NCTD） on cancer stem cells（CSC）. Methods CSC were screened from UMUC3 and Hep G2 cancer cell lines by limiting dilution method. The expression of stemness gene was examined by q PCR. MTS method was used to detect the inhibitory effect of NCTD and paclitaxel（PTX） on CSC. FACS was used to detect the cell apoptosis. Results Sphere-forming cell subpopulations were obtained from two cell lines, with much higher expression of stemness genes NANOG, OCT4, ABCG2, CD133 and CD34. Serum-release led to significant down-regulation of the stemness genes. The CSC was highly resistant to PTX, but the resistance of NCTD was low. NCTD induced apoptosis of the CSC but had no significant effect on the expression of stemness genes. Conclusion NCTD can induce apoptosis of CSC cell, without significant effect on cell differentiation.