紫杉醇长循环纳米胶束联合vMIP-ⅡN端肽逆转乳腺癌细胞耐药的研究

Study on the paclitaxel long-circulating nanomicelles combined with NT21MP to reverse drug resistance in breast cancer MCF-7/PR cells

目的:探讨紫杉醇长循环纳米载药胶束联合病毒巨噬细胞炎性蛋白Ⅱ(vMIP-Ⅱ)N端肽(NT21MP)对乳腺癌紫杉醇耐药细胞株(MCF-7/PR)耐药性的影响。方法:采用磺酰罗丹明B(SRB)染色法检测乳腺癌耐药细胞株MCF-7/PR的存活率;细胞划痕及Transwell实验检测紫杉醇纳米载药胶束联合NT21MP对乳腺癌耐药细胞迁移、侵袭能力的影响;流式细胞术分析细胞凋亡、周期;Western blotting实验检测细胞凋亡、耐药及上皮间充质转化相关蛋白水平表达。结果:相对于紫杉醇组,紫杉醇纳米胶束更有效地抑制细胞增殖、迁移和侵袭,凋亡蛋白Bax、caspase3的蛋白表达水平上调,抗凋亡蛋白Bcl-2表达水平下调,且耐药相关蛋白MRP、P-gp和EMT相关蛋白Vimentin、Slug表达水平下调,E-cadherin表达上调(P<0.01);相对于紫杉醇纳米胶束组,紫杉醇纳米胶束与NT21MP联合组对细胞增殖、迁移、侵袭、细胞周期的抑制,以及抗凋亡和逆转耐药作用更明显(P<0.01)。结论:紫杉醇纳米胶束可有效发挥逆转乳腺癌细胞耐药性的作用,且以紫杉醇纳米胶束联合多肽药物NT21MP作用更明显。

Objective:To investigate the effects of paclitaxel long-circulating nanomicelles combined with vMIP-Ⅱ-N-terminal peptide(NT21MP)on drug resistance of paclitaxel-resistant brest cancer MCF-7/PR cells.Methods:Sulforhodamine B(SRB)staining was used to detect the survival rate of breast cancer paclitaxel-resistant cell line MCF-7/PR.Cell scratch and Transwell assay were used to detect the effects of paclitaxel nanomicelles combined with NT21MP on the migration and invasion of paclitaxel-resistant cells.Flow cytometry was used to analyze the apoptosis and cell cycle.Western blotting was used to detect the expression of apoptosis,drug resistance and EMT-related proteins.Results:Compared with paclitaxel group,the paclitaxel nanomicelles could effectively inhibit the cell proliferation,migration and invasion,the expression levels of apoptotic proteins Bax and caspase3 were up-regulated,the expression levels of anti-apoptotic proteins Bcl-2 were down-regulated,the expression levels of MRP,P-GP and EMT-related proteins Vimentin and Slug were down-regulated,and the expression levels of E-cadherin were up-regulated(P<0.01).Compared with the paclitaxel nanomicelles group,the paclitaxel nanomicelles combined with NT21MP group had more significant effects on the inhibition of cell proliferation,migration,invasion,cycle,anti-apoptosis and reversal paclitaxel resistance.Conclusions:The paclitaxel nanomicelles can effectively reverse the drug resistance of paclitaxel-resistant breast cancer cells,and the effects of the combination of paclitaxel manomicelles and NT21MP is more significant.