Therapeutic Effect Of Tripeptide Tyroservatide (YSV) On Metastasis Of Tumor And Its Mechanisms

Objective:Metastasis is a very complex biological phenomenon. This study was observed ifYSV can inhibit the metastasis of carcinoma and related metastatic mechanisms wereapproached.Methods:1. Experimental lung metastasis models of mouse melanoma B16-F10 cells inC57BL/6 mice were established. Anti-metastasis effect of YSV was evaluated bycounting metastatic nodules.2. Abdominal cavity metastasis models of human ovary carcinoma SKOV-3 in nudemice were established. Anti-metastasis effect of YSV was evaluated bycalculating the survival time and counting metastatic nodules.3. Liver metastasis models after spleen implantation of human colon carcinomaHT-29 and SW620 cells in nude mice were established. Anti-metastasis effect ofYSV were evaluated by calculating the liver index(liver/body weight×100％).Anti-tumor effect of YSV were observed by calculating the spleenindex(spleen/body weght×100％).4. The models of B16-F10 and HT-29 cells transplanted into subcutaneous tissues ofmice were established and the anti-tumor effect of YSV were evaluated.5. In vitro, B16-F10, SKOV-3, HT-29 and SW620 cells were cultured. The effect ofYSV on tumor cells proliferation were observed by MTT method.6. In vitro, B16-F10, SKOV-3, HT-29 and SW620 cells were cultured. The effect ofYSV on tumor cells adhesion to matrigel were observed by MTT method. Theeffect of YSV on tumor cells invasion were observed by Transwell model.7. The effects of YSV on ICAM-1 protein expression in B16-F10 cells and it's lungmetastatic tumor were observed by immunohistochemical staining. The effects of YSV on ICAM-1 protein expression and mRNA level in B16-F10 cells wereobserved by Westem Blot and Real-time PCR.8. The effects of YSV on MMP-2 and MMP-9 protein activities of SKOV-3, HT-29and SW620 cells by zymography. The effects of YSV on MMP-2 and MMP-9protein expression in SKOV-3, HT-29 and SW620 cells and their metastatic tumortissues were observed by Western Blot. The effects of YSV on MMP-2 andMMP-9 mRNA level of SKOV-3, HT-29 and SW620 cells and their metastatictumor tissues were observed by Real-time PCR.Results:1. YSV (640μg/kg/d) significantly inhibited the lung metastasis of B16-F10 cellsimplanted in C57BL/6 mice. The metastasis inhibition rate is 62.21％. The resultof HE staining of lung metastatic lesions display that YSV reduced the size ofmetastatic lesions.2. YSV (640μg/kg/d, 320μg/kg/d) significantly prolonged survival time ofabdominal cavity metastasis model of SKOV-3 in nude mice and the highestsurvival prolong rate is 32.27％; significantly inhibited the number of abdominalcavity metastasis lesions and the highest inhibition rate is 52.93％.3. YSV (640μg/kg/d, 320μg/kg/d) significantly inhibited liver metastasis of HT-29and SW620 cells and inhibited the primary tumor growth of HT-29 and SW620cells.4. YSV (640μg/kg/d,320μg/kg/d,160μg/kg/d) significantly inhibited the growth ofmouse melanoma B16 cells implanted in C57BL/6 mice. The highest inhibitionrate is 54.32％. YSV (640μg/kg/d,320μg/kg/d) significantly inhibited the growthof HT-29 cells implanted in nude mice. The highest inhibition rate is 46.57％.5. YSV significantly inhibited the proliferation of B16-F10, SKOV-3, HT-29 andSW620 cells in vitro. The highest inhibition rates are 50.13％, 42.7％, 64.32％, 63.28％, respectively.6. YSV significantly inhibited B16-F10, SKOV-3, HT-29 and SW620 cells adhesionto matrigel in vitro. The highest inhibition rates are 41.08％, 30.87％, 38.14％,47.06％, respectively. YSV significantly inhibited B16-F10, SKOV-3, HT-29 andSW620 cells invasion in vitro. The highest inhibition rates are 41.52％, 54.55％,62.82％, 46.25％, respectively.9. YSV significantly down-regulated ICAM-1 protein expression in B16-F10 cellsand it's lung metastatic tumor. Significantly down-regulated ICAM-1 mRNAlevel in B16-F10 cells.10. YSV significantly inhibited MMP-2 and MMP-9 protein activities ofSKOV-3, HT-29 and SW620 cells. Significantly down-regulated MMP-2 andMMP-9 protein expression and mRNA level in SKOV-3, HT-29 and SW620 cellsand their metastatic tumor tissues.Conclusion:YSV down-regulated the mRNA level and protein expression of ICAM-1, MMP-2and MMP-9; inhibited the adhesion, invasion and proliferation of tumor cells. Soinhibited the lung metastasis of melanoma, the abdominal cavity metastasis of ovarycarcinoma, the liver metastasis of colon carcinoma.