Effects Of Ulinastatin And Taxotere On Breast Cancer Proliferation, Invasion And VEGF-C, BFGF, NGF Expression

PART ONE EFFECTS OF ULINASTATIN AND TAXOTERE ON BREAST CANCER CELL PROLIFERATION, INVASION AND EXPRESSION OF VEGF-C mRNA, bFGF mRNA, NGF mRNAObjective: To observe the effects of ulinastatin (UTI) and taxotere (TXT) on proliferation, invasion and expression of VEGF-C mRNA, bFGF mRNA, NGF mRNA in breast cancer cell line MDA-MB-231(ER-).Methods: The breast cancer cell line MDA-MB-231 in vitro was randomly divided into four groups: control group, UTI group, TXT group and UTI+TXT group. Expression level of VEGF-C mRNA, bFGF mRNA, NGF mRNA were detected by real time quantitative PCR method. The cell proliferation was analyzed by the MTT assay, and the cell invasion ability was assessed by Transwell chamber.Results: Compared with the control group, the expression level of VEGF-C mRNA, bFGF mRNA, NGF mRNA in MDA-MB-231 cell were significantly down-regulated by UTI and TXT which were used separately or together(p<0.05 for all).The inhibition effect of UTI,TXT and UTI+TXT on expression of the VEGF-C mRNA, bFGF mRNA, NGF mRNA in MDA-MB-231 cell were UTI+TXT>TXT>UTI (p<0.05 for all); except for NGF mRNA, there was no statistically difference between TXT group and UTI+TXT group(p=0.055). In comparison with those of the control group, UTI and TXT which were used separately or together significantly inhibited the proliferation of MDA-MB-231 cell in a time-dependent manner(P<0.05 for all),and the inhibition effect were UTI+TXT>TXT>UTI (p<0.05 for all). UTI and TXT which were used separately or together significantly inhibited the invasion ability of MDA-MB-231 cell (p<0.05 for all), and the inhibition effects were UTI+TXT>TXT>UTI (p<0.05 for all).Conclusion: Both UTI and TXT can significantly inhibit the proliferation, invasion and expression of VEGF-C mRNA, bFGF mRNA, NGF mRNA in breast cancer cell line MDA-MB-231, and a UTI + TXT combination has an additive effect that is superior to TXT alone. A possible mechanism to explain this is that UTI down-regulates the expression of VEGF-C mRNA, bFGF mRNA, NGF mRNA. PART TWO EFFECTS OF ULINASTATIN AND TAXOTERE ON THE GROWTH OF XENOGRAFT BREAST CANCER AND EXPRESSION OF VEGF-C, bFGF AND NGF IN CANCERSObjective: To evaluate the effects of ulinastatin (UTI) and taxotere (TXT) on the growth of mouse xenograft breast tumors and the expression of VEGF-C, bFGF, NGF proteins in breast cancer.Methods: MDA-MB-231 breast cancer cells (ER-)were xenografted into 50 nu/nu nude mice in order to construct a breast cancer xenograft nude mouse model. 21 days after grafting, 29 xenografts in nude mice which met the experimental conditions required were randomly divided into four groups:Control group(n=8), UTI group(n=7), TXT group(n=7), UTI+TXT group (n=7). Mice in each group were injected i.p. for 20 days with physiological saline once per day (control), UTI 1600U/mouse once per day, TXT 20mg/kg once every 7 day, or the combination of UTI with TXT(the same as UTI and TXT, respectively). 20 days later, all survival mice were sacrificed, and treatments were performed as follows: (1) the mean tumor volume of mice in each group was measured; (2) expression of VEGF-C, bFGF, NGF protein in breast cancer tissues was detected by immunohistochemical assay, respectively.Results: According to the growth curve of xenograft tumors: the xenograft tumors volume couldn't be reduced by UTI, but were put off to increase. The difference had statistical significance between UTI group and control group (p<0.05). In comparison with mice in the control group, mice in both TXT group and UTI+TXT group had significantly reduced tumor volume (P<0.05). Compared with TXT group,the xenograft tumors volume in UTI+TXT group had significantly reduced(P<0.05), and there was an additive effects between UTI and TXT for inhibiting the growth of xenograft tumors. In comparison with those of the control group, protein levels of VEGF-C, bFGF and NGF in all treatment groups were reduced significantly (P<0.05 for all), and the inhibition effect for VEGF-C, bFGF, NGF protein expression were UTI+ TXT >TXT>UTI (P<0.05 for all).Conclusions: Both UTI and TXT can significantly inhibit the growth of xenograft tumors and the expression of VEGF-C, bFGF, NGF proteins, and the UTI+TXT combination exhibited an additive effect that is superior to TXT monotherapy. As a result, We suggest a possible mechanism in which ulinastatin down-regulates VEGF-C, bFGF, NGF proteins and thus suppresses the growth of xenograft tumors.