The Molecular Mechanism And Synergistic Effect Of Anti-bFGF Monoclonal Antibody Combined Taxol To Suppress MCF-7 And MCF-7/Taxol

[Objective]: 1. Research the mechanisms of anti-b FGF Mab combined with Taxol to inhibit MCF-7 cell proliferation and induce apoptosis. 2. Preliminary exploration of significance and mechanisms of anti-b FGF Mab combined with Taxol on the drug-resistant strains MCF-7/Taxol.[Methods]: First prepared ascitic type of anti-b FGF monoclonal antibody in vitro cultured breast cancer cells MCF-7 and drug-resistant strains of paclitaxel-resistant MCF-7/Taxol, the experiments are divided into four groups: control group, anti-b FGF monoclonal antibody group, Taxol group and anti-b FGF monoclonal antibody + Taxol group treated with corresponding medicaments for cell processing.(1) CCK-8 assay detected the inhibition rate of cell proliferation anti-b FGF monoclonal antibody combined with Taxol on MCF-7, MCF-7/Taxol cells, and compared the inhibition rates between the four groups.(2) Using flow cytometry Annexin V-FITC / PI double- Staining Detection of MCF-7, MCF-7/Taxol cells in the case of the group treated with different drugs under apoptosis.(3) Using Western blot analysis and MCF-7 and MCF-7/Taxol proliferation and expression of apoptosis-related signaling pathway proteins to reveal b FGF monoclonal antibody combined with Taxol molecule that inhibits the mechanism for MCF-7 and MCF-7/Taxol synergies. Using Western blot to detect the expression changes of protein in Raf-1, Erk, p-Erk, JNK, p-JNK, STAT3, p-STAT3, Caspase3, cleaved-Caspase3, PARP, cleave-PARP, Bax, Bcl.[Results]:(1)CCK-8 showed that the proliferation inhibition rates of Mab D9 combined with Taxol on MCF-7, MCF-7/Taxol was increased by 13.40%-36.50%; 9.56%-26.87%.(2)Flow cytometry showed that the PI, FITC double positive rates of Mab D9 combined Taxol on MCF-7, MCF-7/Taxol were increased by 17.10%-23.20%; 22.00%-22.70%.(3)Western Blot results showed that the combined group can be significantly reduced the expression of Raf-1, p-Erk, p-STAT3, Bcl-2, Caspase3 and increase the expression of p-JNK, Bax, cleaved Caspase3, cleaved PARP’s in MCF-7, MCF-7/Taxol. Raf-1 decreased significantly in MCF-7/Taxol compared with MCF-7.[Conclusion]:(1) This study showed that the proliferation inhibition rates of Mab D9 combined with Taxol on MCF-7, MCF-7/Taxol was increased by 13.40%-36.50%; 9.56%-26.87%. And the PI, FITC double positive rates of Mab D9 combined Taxol on MCF-7, MCF-7/Taxol were increased by 17.10%-23.20%; 22.00%-22.70%.(2)This study reveals the mechanism of anti-b FGF monoclonal antibody combined with Taxol suppressing MCF-7 as follows: strengthening the inhibition of Erk/MAPK pathway, and strengthen raised the expression of p-JNK, and reduced the expression of p-STAT3, causing the expression of Bcl-2 down, Bax upregulation, and the expression changes of Caspase3, cleaved-Caspase3, cleaved-PARP induce apoptosis.(3)The molecule mechanism of synergistic effect about anti-b FGF monoclonal antibody combined with Taxol suppressing paclitaxel-resistant strains(MCF-7/Taxol) are basically the same with the mechanism of and role of parent strain MCF-7, also inhibited Erk / MAPK pathway and raised p-JNK, reduced the expression of p-STAT3, causing down-regulated expression of Bcl-2, upregulated the expression of Bax, Caspase3, cleaved-Caspase3, cleaved-PARP induce apoptosis, anti-b FGF monoclonal antibody and Taxol were effected on MCF-7/Taxol respectively can not cause the same mechanism. And this study found that the expression of Raf-1 was significantly lower in MCF-7/Taxol compared with MCF-7, Raf-1 is likely related with the resistance mechanisms about MCF-7/Taxol resistant to paclitaxel.