羟喜树碱对黑素瘤B16F10细胞侵袭和凋亡的影响及作用机制

Effects and mechanism of hydroxycamptothecin on the invasion and apoptosis of melanoma B16F10 cells

目的:观察羟喜树碱(hydroxycamptothecin,HCPT)对黑素瘤B16F10细胞侵袭能力以及凋亡相关蛋白p53和Bcl-2表达的影响,并探讨其抑制黑素瘤细胞的作用机制。方法:分别用5、10、20、50μmol/L的HCPT处理B16F10细胞24 h及48 h,采用Transwell实验评价细胞的侵袭能力。采用Western blot和RT-PCR检测p53和Bcl-2蛋白表达和mRNA表达。结果:Transwell实验结果显示,与空白组比较,HCPT不同浓度组(除HCPT 5μmol/L组外)B16F10细胞体外侵袭能力明显下降(P<0.01)。HCPT能显著升高p53的mRNA和蛋白水平,同时降低Bcl-2的mRNA和蛋白水平(P<0.01),并呈现出明显的剂量依赖性和时间依赖性。结论:HCPT能够明显降低B16F10细胞的体外侵袭能力,干预B16F10细胞中p53和Bcl-2的表达,从而促进肿瘤细胞凋亡。

Objective:To observe the effects of hydroxycamptothecin(HCPT)on the invasion of melanoma B16F10 cells and the expression of apoptosis-related proteins p53 and Bcl-2,and also to explore possible mechanism of the inhibition of melanoma cells.Methods:Melanoma B16F10 cells were respectively treated with 5,10,20 and 50μmol/L HCPT for 24 and 48 hours.Cell migration was assessed by Transwell assay and protein expression and mRNA expression of p53 and Bcl-2 were detected by Western blot and RT-PCR.Results:Transwell results showed that the in vitro invasion of B16F10 cells significantly decreased in different HCPT concentrations groups(except in 5μmol/L concentration group),as compared with the blank group(P<0.01).In addition,HCPT could significantly increase the mRNA and protein level of p53,decrease the mRNA and protein level of Bcl-2(P<0.01),and at the same time presented a trend of obvious dose-dependence and time-dependence.Conclusion:HCPT could significantly reduce the invasion of B16F10 cells in vitro and influence the expression of p53 and Bcl-2 in B16F10 cells,thus promoting the apoptosis of tumor cells.