肿瘤抑素抗肿瘤相关肽对人卵巢癌细胞作用的体外研究

Effects of antitumor peptide of tumstatin on human ovarian cancer cells in vitro

目的探讨肿瘤抑素(Tumstatin)185-203位氨基酸(19肽)对卵巢癌细胞(SKOV3)的治疗作用。方法使用MTS、Annexin/PI和蛋白印迹的方法检测肿瘤抑素19肽对SKOV3细胞增殖和凋亡的影响,以及Bcl-2、BAX等凋亡相关蛋白的表达情况。检测了19肽对SKOV3细胞侵袭能力的影响,同时应用RT-PCR的方法检测基质金属蛋白-2(matrix metalloproteinase-2,MMP-2)、组织金属蛋白酶抑制物-2(tissue inhibitors of metalloproteinase-2,TIMP-2)和膜型基质金属蛋白-1(membrane-type 1 matrix metalloproteinase,MT1-MMP)mRNA表达量的变化情况。结果19肽能够有效抑制SKOV3细胞的增殖,诱导细胞凋亡,而降低Bcl-2的表达,激活caspase通路可能是其诱导细胞凋亡的分子机制之一。此外该19肽能够有效抑制SKOV3细胞的侵袭,这与抑制MT1-MMP的表达进而抑制MMP-2的膜结合活性相关。结论肿瘤抑素19肽可以通过诱导凋亡和抑制细胞侵袭对体外培养的SKOV3产生杀伤和抑制作用。

Objective To study the biological activity of antitumor poptide of tumstatin （185- 203 amino acids, 19 peptide） in human ovarian cancer cells （SKOV3）.Methods The effects of 19 peptide on proliferation and apoptosis of SKOV3 cell were assayed with MTS and annexin/PI test. The expression of Bcl-2, Bax, caspase 3 and caspase 9 were detected by western blot. Meanwhile, the inhibition of invasion by 19 peptide was observed by cell invasion assay kit. The expression of MMP-2（matrix metalloproteinase-2）, TIMP-2 （tissue inhibitors of metalloproteinase-2） and MT1-MMP（membrane-type 1 matrix metalloproteinase）mRNA were detected by semi-quantltatlve reverse transcription-polymerase chain reaction （RT-PCR）.Results Studies showed that 19 peptide obviously not only inhibited proliferation and induced apoptosis of SKOV3, but also reduced the expression of Bcl-2 and activated caspase pathway. Furthermore, 19 peptide inhibited both the invasion of SKOV3 cells as well as the activation of membrane-bound MMP-2 by decreasing the expressions of MT1-MMP. Conclusion The antitumor peptide of tumstatin has potential to inbibit ovarian cancer cell proliferation and invasion in vitro.