肿瘤抑素7肽对小鼠黑色素瘤细胞B16生长和凋亡的影响

The Effect of Tumstatin 7 Peptide on Proliferation and Apoptosis of B16 Cell

目的研究肿瘤抑素7肽(CNYYSNS)对小鼠黑色素瘤细胞B16增殖和凋亡活性的影响。方法四甲基偶氮唑蓝法、生长曲线法观察7肽对小鼠黑色素瘤B16细胞增殖抑制作用;TUNEL法、HE染色和透射电镜观察7肽对小鼠黑色素瘤细胞B16形态影响,并以人脐静脉内皮细胞(ECV304)为对照细胞,观察7肽对非肿瘤细胞增殖的抑制作用。结果 7肽可显著抑制小鼠黑色素瘤细胞B16增殖,且呈剂量、时间依赖性,其半数抑制浓度(IC50)为8.53×10-5mol·L-1;TUNEL法测定细胞凋亡率为68.45%;电镜观察证实实验组可见明显凋亡细胞,核仁固缩。而7肽对人脐静脉内皮细胞ECV304作用较弱,IC50为5.78×10-4mol·L-1,细胞形态变化不明显。结论肿瘤抑素7肽可抑制小鼠黑色素瘤B16细胞生长,促进其凋亡,对黑色素瘤的治疗具有潜在价值,而对增殖速度较快的内皮细胞影响较小,说明7肽对肿瘤的抑制作用具有一定特异性。

OBJECTIVE To study the effect of biological activity of tumstatin 7 peptide （CNYYSNS） on the cell proliferation and apoptosis of B16 melanoma cell. METHODS The inhibitory effect of tumstatin 7 peptide on the proliferation of B16 cell was observed by MTr and cell growth curves. The influence of tumstatin 7 peptide on morphology of B16 cell was perceived by TUNEL, HE staining and the transmission electron microscope （TEM）. Human umbilical vein endothelial cell （ ECV304 ） as control cell was detected that tumstatin 7 peptide affected the proliferation of non-tumor cells. RESULTS Tumstatin 7 peptide can significantly inhibit the prolifera- tion of B16 cell in dose- and time-dependent manner. Its IC50 was 8.53×10^-5mol·L-1 , The mophology of B16 cell was obviously changed by means of TUNEL assay, HE staining and TEM. They appeared karyopyknosis and apoptotic bodies. The apoptosis rate of B16 cell was 68.45%. The effect of 7peptide on human endothelial cell was weak, its IC50 was 5.78×10～mol·L-1. CONCLU- SION Tumstatin 7 peptide can inhibit the proliferation of B16 cell and promote B16 cell apoptosis. It has little effect on endothelial cell, which revealed 7 peptide having a certain specificity of anti-tumor. It will be of great potential value to melanoma treatment.