Effect Of Ulinastatin And Cyclophosphamide On Matrix Metalloproteinase-9 Expression And Proliferation Invasion Of Breast Cancer

PART 1 EFFECT OF ULINASTATIN AND CYCLOPHOSPHAMIDE ON MATRIX METALLOPROTEINASE-9 EXPRESSION, PROLIFERATION AND INVASION IN BREAST CANCER CELL LINE MDA-MB-231 AND MCF-7Objective: To observe the effect of ulinastatin (UTI) and Cyclophosphamide on matrix metalloproteinase-9 expression, proliferation and invasion in breast cancer cell line MDA-MB-231 and MCF-7.Methods: The breast cancer cell line MDA-MB-231 and MCF-7 were randomly divided into eight groups: UTI low dose, UTI middle dose, UTI high dose, CTX, UTI low dose+ CTX , UTI middle dose+ CTX, UTI high dose+ CTX, and blank control group. The cell proliferation was analyzed by the MTT assy. Expression of MMP-9 was detected by reverse transcription and quantitative real-time polymerase chain reaction (Real time RT-PCR). And the invasion ability of MDA-MB-231 MCF-7 cells was tested by Boyden chamber assay.Results: Compared with the control group,the UTI significantly inhibited the proliferation of both ER-positive MCF-7 and ER-negative MDA-MB-231 cells induced by CTX in a dose-dependent pattern. The expression of MMP-9 in MDA-MB-231 and MCF-7 cells was down-regulated and the invasion ability of which was decreased after the treatment. Compared with the CTX group, the inhibition effect on the proliferation was weaker, but the inhibitory efficient of UTI combined with CTX was higher than that of CTX used alone.Conclusions: UTI has a synergetic effect on cell apoptosis induced by CTX in in breast cancer cell line MDA-MB-231 and MCF-7, the down-regulation of MMP-9 expression may be one of the main mechanisms. PART 2 INHIBITION OF ULINASTATIN AND CYCLOPHOSPHAMIDE ON MATRIX METALLOPROTEINASE-9 EXPRESSION IN NUDE MOUSE WITH MCF-7 BREAST CANCERObjective: To investigate the inhibition of Ulinastatin and Cyclophosphamide on Matrix Metalloproteinase-9 expression in nude mouse with MCF-7 breast cancer.Methods: Choosing Urinary trypsin inhibitor (UTI), breast cancer model of nude mouse; 40 female nude mouse were inoculated with human breast cancer cell MCF-7, then grouping them to normal sodium (control group), CTX,UTI 2.5*104,UTI 5*104,UTI 10*104,CTX+UTI 2.5*104,CTX+UTI 5*104 and CTX+UTI 10*104, there were 5 in each group, 7days after innoculation, injecting drugs in abdominal cavity, 14 days after innoculation we killed all the mouse, then adding up the average weight of the hypodermatic tumor, analyzing the expression of MMP-9 by immunohistochemistry, detecting the effect of UTI and CTX on MMP-9 in breast cancer by RT-PCR.Results: The different weight of hypodermatic tumor was (7.58±0.57,1.21±0.11*, 3.78±0.42*, 3.01±0.38*, 1.97±0.13*, 0.83±0.12*, 0.72±0.11* and 0.43±0.063*)g, (*compared with that of control group: P<0.05), expression of MMP-9 protein, that of UTI and CTX compared with control group decreased (P<0.05), UTI+CTX compared with CTX: Expression of protein decreased (P<0.05), and UTI,CTX compared with control group, expression of MMP-9 gene decreased (P<0.05), UTI+CTX compared with CTX, expression of MMP-9 gene decreased too (P<0.05).Conclusion: UTI and CTX can both inhibit the proliferation of breast cancer, and they had synergistic effect. The mechanism might be concerned with the descending expression of MMP-9 protein and gene by UTI.