Inhibitory Effect Of Endostar Combined With Paclitaxel-cisplatin On VEGF And MMP-2Expressions Of Human Breast Cancer Cell MCF-7in Xenograft-bearing Nude Mice

ObjectiveInvestigate the tumor vascular endothelial growth factor (VEGF) andmatrix metalloproteinase-2(MMP-2) expressions of endostar,paclitaxel-cisplatin (TP), endostar combined with TP on human breast cancercell MCF-7in xenograft-bearing nude mice measured by immunefluorescencedouble staining and Western blot; then evaluate the anti-angiogenic effect ofendostar, the invasion and metastasis of tumor and the synergistic effect of TPregimen with endostar regimen.Methods1. Take the frozened MCF-7cellline from the liquid nitrogen, dissolveMCF-7cellline in37℃water bath, then MCF-7cellline grown in DMEMmedium which contained fetal calf serum, the cells grown adherent, passageculture1time in2~3d, the logarithmic growth phase MCF-7cells werepercussed into cell suspension,1×107/0.2ml, inject0.2ml on the right breastpads in the nude mice, then24xenograft nude mice were randomized into4groups and received treatment accordingly: control (saline), TP, endostar andcombination therapy (endostar+TP). The tumor inhibition rate and tumorgrowth curves were measured before, during and after2weeks’ treatment foreach group.2. After2weeks’ treatment, nude mice in each group were killed bycervical dislocation, taken some tumor specimens, a part of tumor tissue were handled with alcohol, embedded in xylene and paraffin, serial section in5μm,the rest were frozen in liquid nitrogen at-80℃. VEGF and MMP-2expressionlevels were detected using immunefluorescence double staining and Westernblot. The results were analysed by statistics.Results1. After treatment, xenograft tumor volumes in endostar group and thecombination group were significantly decreased than the control group (p<0.01), the tumor growth curve were gently rising. Neoadjuvant chemotherapygroup and the control group did not have a statistically difference in tumorvolume (p>0.05), the tumor growth curves were upward trend.2. Immunofluorescence double staining showed that: VEGF and MMP-2expression levels in neoadjuvant chemotherapy group, endostar group, thecombination group were significantly lower than the control group, thedifference was statistically significant (p<0.01); neoadjuvant chemotherapygroup and endostar group did not have a statistically difference in VEGFexpression level (p>0.05).3. Western blot showed that: VEGF and MMP-2expression levels inneoadjuvant chemotherapy group, endostar group, the combination groupwere significantly lower than the control group, the difference was statisticallysignificant (p<0.05).4. VEGF and MMP-2expression levels in the combination group weresignificantly lower than neoadjuvant chemotherapy group and endostar group,the control group, the difference was statistically reduced. TP regimen had asynergistic effect with endostar regimen in anti-angiogenesis when VEGFexpression was detected using immunefluorescence double staining andWestern blot (p<0.05). TP regimen had a synergistic effect with endostarregimen in anti-angiogenesis when MMP-2expression was detected usingimmunefluorescence double staining (p<0.05). Conclusion1. Endostar, as a drug targeting inhibit tumor angiogenesis, invasion andmetastasis, can reduce the expression level of VEGF and MMP-2in breastcancer tissue. TP regimen also has a certain degree of anti-angiogenesis andinhibition effect of invasion and metastasis.2. TP regimen has a synergistic effect with endostar regimen, can improvethe therapeutic efficacy to some extent, then VEGF and MMP-2expressionsreduced significantly, thus clearly showing anti-angiogenesis and the inhibitionof tumor invasion and metastasis. So it would provide a theoretical basis forthe evaluation of therapeutic efficacy in human breast cancer.