rSIFN-co对乳腺癌MCF-7/ADR细胞增殖、凋亡和表柔比星耐药性的影响

Effect of recombinant super-compound interferon on proliferation,apoptosis and resistance to epirubicin of human breast cancer cell MCF-7 /ADR

目的:观察重组复合高效干扰素(recombinant super-compound interferon,rSIFN-co)在体外对多药耐药(multi-drug resistance,MDR)的人乳腺癌MCF-7/ADR细胞的增殖、凋亡和表柔比星耐药性的影响,并探讨其可能的作用机制。方法:分别使用rSIFN-co、表柔比星及rSIFN-co联合表柔比星处理MCF-7/ADR细胞,以MCF-7细胞作为对照,MTT法和流式细胞术分别检测rSIFN-co对MCF-7/ADR细胞增殖、凋亡的影响,免疫细胞化学方法检测rSIFN-co对MCF-7/ADR细胞中P-gp表达水平的影响。结果:各组药物作用24 h后,0.078μg/ml rSIFN-co单独作用和0.02μg/ml rsIFN-co联合15.00μg/ml表柔比星对MCF-7/ADR细胞体外生长的抑制率即显著高于100.00 g/ml的表柔比星[(29.7±1.4)%、(23.0±2.1)%vs(17.1±1.5)%,均P<0.01],各组药物对MCF-7/ADR细胞的抑制率呈时间、浓度依赖性;rSIFN-co联合表柔比星作用72 h后表现出协同作用。表柔比星作用24 h后,MCF-7/ADR细胞凋亡率与对照组相比无显著变化(P>0.05);而rSIFN-co单用或联合表柔比星作用24 h后,凋亡率即较单用表柔比星组显著增加[(35.37±1.40)%、(61.37±1.76)%vs(9.80±1.66)%,均P<0.01],其促凋亡的作用呈时间依赖性;并且rSIFN-co与表柔比星具有协同作用。表柔比星组P-gp的表达较对照组显著升高[(4.17±0.0252)vs(3.94±0.0088),P<0.01],rSIFN-co组与联合组P-gp的表达均显著下调[(2.59±0.0260)、(2.62±0.0100)vs(3.94±0.0088),均P<0.01);联合组与单用rSIFN-co相比无显著差异(P=0.948)。结论:rSIFN-co能够抑制MCF-7/ADR细胞增殖并促进其凋亡,同时可能通过下调P-gp蛋白的表达来增加其对表柔比星的敏感性。

Objective ：To observe the effect of recombinant super-compound interferon （rSIFN-co） on the proliferation, apoptosis and resistance to epimbicin of human breast cancer MCF-7/ADR ceils （a multi-drug resistance [ MDR ] strain）, and to investigate the possible mechanism. Methods： MCF-7/ADR cells were treated with rSIFN-co, epirubicin alone or combaination, and the MCF-7 cells were used as control. MTF assay and flow cytometry were performed to detect the effect of rSIFN-co on the proliferation and apoptosis of MCF-7/ADR cells, respectively. Immunohistochemical staining was used to detect the influence of rSIFN-co on the P-gp expression level in MCF-7/ADR ceils. Results ： After treated by different drugs for 24 h, the growth inhibition rate of MCF-7/ADR cells treated by 0. 078 ug/ml rsIFN-co or 0.02 ug/ml rsIFN-co combined with 15.00 ug/ml epirubicin was significantly higher than that treated by 100.00 ug/ml epirubicin （ [29.7 ± 1.41%, [23.0 ±2.1 ] % vs [ 17.1 ± 1.5]% ; all P 〈0.01）. The inhibition effect of each drug had a dose and time dependence. Synergistic effect of rSIFN-co with epirubicin was also observed after being treated for 72 h. Epiru bicin showed no significant effect on MCF-7/ADR cells＇ aooptosis after treated for 24 h （ P 〉 0.05 ） ： however, use ofrSIFN-co alone or combined with epirubicin significantly enhanced the apoptosis rate than did epirubicin alone after 24 h （[35.37± 1.40]%, [61.37 ± 1. 761% vs [9.80 ± 1. 661% ; all P 〈0.01）, and the effects on cell apoptosis had a time dependence （P 〈 0.01 ） ; and the synergistic effect of rSIFN-co with epirubicin was also observed. Compared with the control group （3.94 ± 0. 0088 ）, the P-gp expression was increased in the epirubicin group （4.17 ±0. 0252, P 〈 0.01 ）, but decreased in rSIFN-co group （2.59 ± 0. 0260, P 〈 0.01 ） and the combined group （2.62 ± 0. 0100, P 〈 0.01 ）. There was no significant difference between the combined group and rsIFN-co group in P-gp expression （ P = 0. 948 ）. Conclusion： rSIFN-co can inhibit cell growth, induce cell apoptosis of human breast cancer MCF-7/ADR cells, and re verse the multi-drug resistance by decreasing the expression of P-gp.