丹参酮ⅡA诱导人黑素瘤细胞A375自噬的小鼠模型研究

Discussion on Inhibitory Effect Mechanism of Tan ⅡA on the Proliferation of Melanoma Cells A375 Based on Autophagy

目的探讨丹参酮ⅡA是否通过诱导自噬的作用机制抑制人黑素瘤细胞A375的增殖。方法将人黑素瘤细胞A375制备的模型小鼠24只随机分为对照组、紫杉醇组(尾静脉注射,8mg/kg,隔天1次,持续28天)和丹参酮ⅡA低、高剂量组(25mg、50mg 1次/d,持续28天),每组6只。实验结束后,随即处死小鼠,手术剥取瘤块称重。采用RT-PCR测定小鼠瘤组织中自噬相关基因Beclin-1、微管相关蛋白1轻链3(LC3)-ⅡmRNA水平;Western印迹法检测Beclin-1、LC3-Ⅱ蛋白的表达水平。结果丹参酮ⅡA可显著降低人黑素瘤细胞A375制备的模型小鼠肿瘤质量系数,抑瘤率达56.70%;可显著上调瘤组织中自噬相关基因Beclin-1、LC3-ⅡmRNA及其蛋白的表达,较模型组差异均有统计学意义(P<0.05)。结论丹参酮ⅡA通过诱导人黑素瘤细胞A375的自噬机制,从而抑制其增殖,延缓肿瘤生长。

Objective To investigate whether Tan Ⅱ A inhibits the proliferation of melanoma ceils A375 through inducing autophagy. Methods Twenty-four mouse models were established using human melanoma ceils and randomly divided into control group, paclitaxel group（ tail vein injection, 8mg/kg, every other day for 28 days）, Tan Ⅱ A low-dose and high-dose groups （25mg,50mg, for 28 days）, with 6 mices in each group. At the end of the experiment, the mices were sacrificed and the tumors were excised and weighed. The mRNA expression of autophagy related genes, such as Be- clinl and microtubule-associated protein 1 light chain 3 （LC3）- Ⅱ were measured by RT-PCR. The protein expression of Beclin-1 and LC3-Ⅱ were detected by western blot. Results Tan Ⅱ A could significantly reduce tumor weight coefficient of melano- ma cells A375 model mices, with tumor inhibitory rate of 56. 70% ; it also signifi- cantly up-regulated the mRNA and protein exression of Beclin-1 and LC3-Ⅱ , with statistical significance compared to the control group （P 〈 0. 05 ）. Conclusion Tan ⅡA can inhibit the proliferation of melanoma cells A375 and slow down the growth of tumor through inducing the mechanism of autophagy.