摘要
目的探讨平阳霉素抑制ECV304细胞生长的分子机制。方法采用流式细胞术测定ECV304细胞的天冬氨酸特异性半胱氨酸蛋白酶-3(cysteinyl aspartatespecific protease-3,Capase-3)、p53、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)蛋白表达水平在平阳霉素作用后的变化;采用TRAP-银染法检测ECV304细胞的端粒酶活性。结果平阳霉素可导致ECV304的caspase-3表达水平增高,但与浓度无关;ECV304的p53荧光强度在平阳霉素75μg/ml和150μg/ml随浓度的增加而提高,平阳霉素作用后ECV304的Bcl-2,ECV304细胞经15μg/ml平阳霉素作用后端粒酶表达减弱,75、150μg/ml组端粒酶活性被抑制。结论平阳霉素对体外培养的人脐静脉内皮细胞系ECV304细胞的抑制作用主要是通过诱导细胞凋亡来实现的。
OBJECTIVE The aim of this study was to investigate the molecular mechanism of Pingyangmycin in inducing the apoptosis of human umbilical vein endothelial cells(ECV304). METHODS ECV304 cells were cultured and passaged, and then were divided into one control group and 3 treatment groups. The 3 treatment groups were treated with 15, 75 and 150 μg/ml Pingyangmycin for 24h respectively. The expressions of caspase-3, p53 and Bcl-2 in ECV304 cells were detected by flow cytometry, and the activity of telomerase was determined using telomere repeat amplification protocol(TR AP) silver staining method. RESULTS After treatment with different dose of Pingyangmycin, the expression of Caspase-3 in ECV304 cells was increased, but no d o s e i nd e p e nd e nt. P i ng y a ng myc i n c ou ld significantly increase the expression of p53 with dose dependence. It had no obvious effect on Bcl-2 expression. There was high expression of telomerase in control group. After treatment with different dose of Pingyangmycin, the telomerase activity was significantly decreased.CONCLUSION Pingyangmycin can increase Caspase-3 and p53 expression levels and inhibit telomerase activity, that induce the apoptosis of ECV304.
出处
《中国耳鼻咽喉头颈外科》
CSCD
2014年第6期304-306,共3页
Chinese Archives of Otolaryngology-Head and Neck Surgery
