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目的研究bFGF单抗联合紫杉醇(Taxol)抑制MCF-7细胞增殖、诱导细胞凋亡的分子机制;探讨bFGF单抗联合Taxol对耐药株MCF-7/Taxol的增效作用及机制。方法 CCK-8法检测bFGF单抗联合Taxol对MCF-7及MCF-7/Taxol增殖抑制率;流式细胞术检测二者联合对MCF-7、MCF-7/Taxol的凋亡作用;Western blot检测MCF-7及MCF-7/Taxol相关信号通路蛋白的表达变化。结果 CCK-8法检测结果显示bFGF单抗联合Taxol作用MCF-7、MCF-7/Taxol后对其增殖抑制率分别提高了13.40%~36.50%、9.56%~26.87%,联合能加强对细胞抑制作用。流式细胞术结果显示bFGF单抗联合Taxol作用MCF-7、MCF-7/Taxol后PI、FITC双阳性率分别提高了17.10%~23.20%、22.00%~22.70%,表明联合能加强对凋亡作用。Western blot结果显示MCF-7、MCF-7/Taxol中MabD9+Taxol组能明显下调Raf-1、p-Erk、p-STAT3、Bcl-2、Caspase3,并上调p-JNK、Bax、cleavedCaspase3、cleaved PARP,其中Raf-1在MCF-7/Taxol中表达量较MCF-7明显下降。结论本研究揭示了bFGF单抗联合Taxol抑制MCF-7的如下机制:加强对Erk/MAPK通路的抑制,并加强上调p-JNK,下调p-STAT3的表达量,引起Bcl-2表达量下调,Bax表达上调,Caspase3、cleaved Caspase3、cleaved PARP表达变化引起细胞凋亡。且耐药株除具类似的抑制机制外,其Raf-1下调更加明显。
OBJECTIVE: To study the molecular mechanism of bFGF monoclonal antibody combined with paclitaxel (Taxol) on MCF-7 cell proliferation and induce apoptosis. To investigate the synergistic effect and mechanism of bFGF monoclonal antibody combined with Taxol on drug-resistant strain MCF-7 / Taxol. Methods The inhibitory rate of proliferation of MCF-7 and MCF-7 / Taxol induced by bFGF combined with Taxol was detected by CCK-8 assay. The apoptosis of MCF-7 and MCF-7 / Taxol cells was detected by flow cytometry. Western blot The expression changes of MCF-7 and MCF-7 / Taxol related signal pathway proteins were detected. Results The results of CCK-8 assay showed that the inhibition rate of MCF-7 and MCF-7 / Taxol increased by 13.40% -36.50% and 9.56% -26.87%, respectively, effect. The results of flow cytometry showed that the positive rates of PI and FITC were increased by 17.10% -23.20% and 22.00% -22.70% respectively after combination of bFGF monoclonal antibody with Taxol and MCF-7, and MCF-7 / Taxol. effect. Western blot results showed that MabD9 + Taxol group could significantly downregulate Raf-1, p-Erk, p-STAT3, Bcl-2 and Caspase3 in MCF-7 and MCF-7 / Taxol groups and upregulate p-JNK, Bax, cleavedCaspase3 and cleaved PARP, the expression of Raf-1 in MCF-7 / Taxol was significantly lower than that of MCF-7. Conclusion This study revealed that bFGF combined with Taxol inhibits the mechanism of MCF-7 by increasing the inhibition of Erk / MAPK pathway and upregulating p-JNK, down-regulating the expression of p-STAT3 and down-regulating the expression of Bcl- Bax expression upregulation, Caspase3, cleaved Caspase3, cleaved PARP expression changes caused apoptosis. In addition to the resistant strains with a similar inhibition mechanism, the Raf-1 downregulation more obvious.