CD133功能对人脑胶质瘤及恶性黑色素瘤细胞体外生长的影响

Effect of CD133 Function on in vitro Growth of Different Malignant Tumor Cells

目的分析肿瘤干细胞表面标记物CD133/Prominin-1的功能对人脑胶质瘤及恶性黑色素瘤肿瘤细胞体外生长特性的影响。方法选取人脑胶质瘤细胞U251及人恶性黑色素瘤细胞A357为对象,采用脂质体介导CD133/Prominin-1反义寡核苷酸(ASODN)及无关寡核苷酸(NSODN)分别转染细胞。转染后采用RT-PCR检测CD133/Prom-inin-1 mRNA水平,同时接种各组细胞,转染后第14天观察细胞生长情况。结果在A357细胞中,CD133/Prominin-1ASDON转染组的CD133/Prominin-1 mRNA水平明显低于空白对照组及NSODN转染组(P<0.05),但其细胞集落形成率与其它两组比较差异并无统计学意义。在U251细胞中,CD133/Prominin-1 ASDON转染组的CD133/Prominin-1mRNA水平亦低于空白对照组及NSODN转染组,其第14天细胞数量也较其它两组明显减少(P<0.05)。结论通过反义寡核苷酸(ASODN)转染下调CD133/Prominin-1表达对人恶性黑色素瘤细胞的集落形成能力无明显影响,但可以显著抑制人脑胶质瘤细胞的体外增殖。

Objective To investigate whether the function of CD133/Prominin-1,the cancer stem cells（CSCs）marker of glioma and malignant melanoma,can affect the in vitro growth of these two types of tumor cells.Methods Two different cancer cell lines-a human glioma cell line and a human malignant melanoma cell line were transfected with CD133/Prominin-1 antisense oligodeoxynucleotide（ASODN）or nonsense oligodeoxynucleotide（NSODN）mediated by lipofectin.After transfection,CD133/Prominin-1 mRNA expression level was detected by RT-PCR.The cells in each group were collected and planted,and the in vitro growth of both two cell lines was observed.Results In the melanoma cells,the CD133/Prominin-1 mRNA expression level in the ASODN group was apparently lower than in the NSODN group and control group.But there was no significant difference in the colony forming rate between ASODN group and NSODN group,or control group.In the glioma cells,the cell number in the ASODN group was markedly less than in the other two groups.Conclusion The knock-down of CD133/Prominin-1 expression by ASODN transfection can not apparently change the colony forming rate of melanoma cells,but can markedly inhibit the in vitro growth of glioma cells.