赤芍总苷抑制人黑色素瘤A375细胞株增殖作用的研究

Inhibitory Effect of Total Paeonyglucosides on Proliferation of Human Malignant Melanoma A375 Cells

目的观察赤芍总苷(TPG)对人黑色素瘤A375细胞增殖的影响及其作用机制。方法黑色素瘤A375细胞,分别加入12.5、25、50、100、200 mg/L TPG,对照组加等量0.9%氯化钠注射液,孵育48 h。采用四甲基偶氮唑蓝(MTT)方法测定细胞生长抑制率,用RT-PCR、Western blot法分别检测增殖细胞核抗原(PCNA)、P21、P27、P53 mRNA及其蛋白的表达水平。结果不同浓度(12.5、25、50、100、200 mg/L)TPG刺激后,对黑色素瘤细胞均具有明显的增殖抑制作用。100 mg/L TPG作用黑色素瘤细胞48 h,PCNA mRNA及其蛋白表达水平较对照组显著下降,而P21、P27、P53的mRNA及其蛋白表达水平较对照组显著上升,差异有统计学意义(P<0.01)。结论TPG抑制黑色素瘤A375细胞增殖,与下调PCNA表达及上调P21、P27、P53表达有关。

Objective To investigate the effects of total paeonyglucosides （TPG） on proliferation of human malignant melanoma A375 cell, and to further elucidate its molecular mechanisms. Methods Human malignant melanoma cells were cultured with TPG （ 12. 5, 25, 50, 100, 200 mg/L） and the control group was cultured with 0. 9% NaCI injection for 48h. The rates of growth inhibition were tested by MTT assay. RT - PCR and Western blot were used to test PCNA, P21, P27 and P53 mRNA and the expression of protein. Results The proliferation of human malignant melanoma cells was obviously inhibited by TPG of different concentrations （ 12. 5, 25, 50, 100, 200mg/L） . After culturing with 100 mg/L TPG for 48 h, the level of PCNA mRNA and the level of its protein expression in malignant melanoma cells were significantly decreased compared with control group, while the level of mRNA in P21, P27 and P53 and the level of their protein expression were significantly increased （P 〈 0. 01 ） . Conclusion TPG can inhibit human malignant melanoma proliferation by down - regulating the expression of PCNA and up - regulating P21, P27 and P53.