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目的研究赤芍总苷(total paeonyglucosides,TPG)联合化疗药物卡铂(CBP)对人黑色素瘤A375细胞生长的抑制作用及相关机制。方法用TPG和CBP作用于体外培养的黑色素瘤细胞,SRB法检测药物对黑色素瘤细胞生长的作用,逆转录-聚合酶链反应法(RT-PCR)检测各组细胞PCNA、cyclin D、P21 mRNA的表达情况;Western blot法检测各组细胞PCNA、cyclin D、P21蛋白表达水平。结果 CBP处理组OD值较对照组明显降低,TPG联合CBP导致OD值进一步下降(P<0.01);CBP处理组黑色素瘤细胞PCNA和cyclin D mRNA和蛋白水平均较对照组显著降低(P<0.01),而P21 mRNA和蛋白水平则明显升高(P<0.01);TPG联合CBP作用于黑色素瘤细胞,PCNA和cyclin D mRNA和蛋白水平较CBP单独作用组下降更为明显;P21 mRNA和蛋白水平较CBP单独作用组升高更加显著。结论 TPG和CBP联合用药效果优于CBP单独用药,可进一步提高药物对肿瘤细胞的生长抑制,与降低PCNA和cyclin D表达、升高P21表达有关。
Objective To study the inhibitory effect of total paeonyglucosides (TPG) combined with carboplatin (CBP) on the growth of human melanoma A375 cells and its related mechanisms. Methods The cultured melanoma cells were treated with TPG and CBP. The effect of drugs on the growth of melanoma cells was detected by SRB method. The expressions of PCNA, cyclin D and P21 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR) The expression of PCNA, cyclin D, P21 in each group was detected by Western blot. Results Compared with control group, the OD value of CBP group was significantly lower than that of control group (P <0.01). The OD value of CBP group decreased further (P <0.01). The mRNA and protein expressions of PCNA and cyclin D in CBP group were significantly lower than those in control group ), P21 mRNA and protein levels were significantly increased (P <0.01); TPG combined with CBP on melanoma cells, PCNA and cyclin D mRNA and protein levels more significantly decreased compared with CBP alone group; P21 mRNA and protein levels Compared with CBP alone group increased more significantly. Conclusion The combined effect of TPG and CBP is superior to that of CBP alone, which can further increase the inhibition of tumor growth by decreasing the expression of PCNA and cyclin D and increasing the expression of P21.