Stady Of Tumstatin7Peptide Against Tumors

Tumstatin possessed two antitumor active regions, one was T7peptide located in N terminal and composed of74-98amino acid, T7peptide could inhibitd the angiogenesis to play the indirect antitumor action. Another one was19peptide located in C terminal and composed of185-203amino acid, it could inhibit the cancer cell growth, induce the cell apoptosis to direct antitumor. In order to improve the antitumor activity and decrease the side-effect of tumstatin, we reformed the19peptide using the biosynthesis method, and got the tumstatin7with high purity(>98%), smaller molecule weight, which kept the antitumor activity of19peptide.We used MTT assay, cell growth curve, HE staining morphology assay. TUNEL in situ end labeling, flow cytometer and launch electron microscope to investigate the effect of7peptide on cell multiplication and antiapoptosis in vitro.MTT assay results showed that7pepetide could significantly inhibit several tumor cell growth in48h, the IC50of melanoma cell B16. human ansamll lung cancer cell A549, human hepatoma cell HepG-2, human gastric carcinoma SGC7901and ECV304is72.53μg/ml、92.84μg/ml、160.11μg/ml、311.38μg/ml。490.81μg/ml, and submited the good dose dependent. Cell gorwth curve results showed that7peptide100μg/ml could significantly change the curve basic tendency of B16and A549cells, and had little inhibitory effect or HepG-2and SGC7901cells followed by changed basic gorwth curve tedency,but7peptide could only decrease the cell number of HUVEC, can not changed the growth curve tedency. A Series of assay(HE Assay, TUNEL assay, electron macroscope assay) results indicated that there were apparent morphology and color change between experimental group and control group, the most obvious were B16and A549cell, the second were HepG-2and SGC7901cells. and there were no obvious change in ECV304cells. We aslo used TUNEL assay to investigated the7peptide antiapoptosis effect, the results indicated that7peptide could significantly induce the B16and A549cell apoptosis followed by more flourescence, and had little effect on ECV304cells with little flourescence in TUNEL assay.We can see the brown apoptosis cells in meidcation administrtion group under light microscope, the apoptosis rate of7peptide in B16. A549.HepG-2. SGC7901and ECV304is68.45%、54.82%、30.31%、16.06%、10.19%. FCM results showed7peptide also had cycle blocking action in above four kinds of tumor cells except for ECV304cells. We used tranmission electron microscope investigated the effect of7peptide(100ug/ml.48h) on cell ultranstructural organization. a series of distinctive apoptosis morphology change including cell microvillus disappeared, chromatin pycnosis, collection, apoptotic body forming were coming in experimental group, but there was no obvious apoptosis phenomenon in ECV304cells.In conclusion,7peptide possesed double antitumor activity which could inhibit cell proliferation and induce cell apoptosis, Itsor antitumor activity was cell specific, which had strong effect on B16and A549cells, little effect on HepG-2and SGC7901cells, almost no effect on ECV304cells, these results showed that7peptide had no obvious angiogenesis inhibitory effect, little effect on human normal cells. Our investigation confirmed that7peptide had antitumor activity with little side-effect, and could be a new. prospect drug in tumor clinical treatment.