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白藜芦醇牛血清白蛋白纳米颗粒诱导人卵巢癌SKOV3细胞死亡的机制

[Mechanisms of resveratrol bovine serum albumin nanoparticle-induced cell death in human ovarian cancer SKOV3 cells].

作者信息

Guo Li-yuan, Peng Yan, Li Yu-lian, Yao Jing-ping, Wang Jing, Zhang Guang-mei, Chen Jie, Sui Li-hua

机构信息

Third Hospital of Harbin Medical University, Harbin 150081, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2010 Nov;30(11):2440-2.

Abstract

OBJECTIVE

To study the effect of resveratrol bovine serum albumin nanoparticles on SKOV3 cell line and its mechanisms.

METHODS

The morphological changes of the cells exposed to the nanoparticles were observed by apoptotic body/cell nucleus DNA staining under inverted microscope and fluorescence microscope, and the pathway of cell death was determined by phosphatidylserine translocation. Western blotting was performed to detect the activation of cyto.c, caspase-3 and caspase-9.

RESULTS

DNA ladder was detected with gel electrophoresis and the cell death was partially inhibited by the pan-caspase inhibitor Z-VAD-FMK. Gel electrophoresis displayed both DNA ladder and smear in RES-BSANP exposed groups, while DNA ladder disappeared in Z-VAD-FMK group and only the smear was left. Cyto.c in the cytoplasm was released at 2 h, while the expression of caspase-9 protein reached the peak level at 4 h and caspase-3 expression was obvious enhanced at 8 h. At 4 h, caspase-9 expression in the cells exposed to 100 µmol/L RES-BSANP was decreased significantly as compared to the cells treated with 50 µmol/L RES-BSANP (P>0.05).

CONCLUSION

RES-BSANP can induce the necrosis and apoptosis of SKOV3 cells via either caspase-dependent or caspase-independent pathways.

摘要

目的

研究白藜芦醇牛血清白蛋白纳米粒对SKOV3细胞系的作用及其机制。

方法

通过凋亡小体/细胞核DNA染色,在倒置显微镜和荧光显微镜下观察纳米粒作用后细胞的形态变化,采用磷脂酰丝氨酸转位法确定细胞死亡途径。采用蛋白质免疫印迹法检测细胞色素c(cyto.c)、半胱天冬酶-3(caspase-3)和半胱天冬酶-9(caspase-9)的激活情况。

结果

凝胶电泳检测到DNA梯状条带,泛半胱天冬酶抑制剂Z-VAD-FMK可部分抑制细胞死亡。凝胶电泳显示,白藜芦醇-牛血清白蛋白纳米粒(RES-BSANP)作用组出现DNA梯状条带和拖尾现象,而Z-VAD-FMK组DNA梯状条带消失,仅留拖尾现象。细胞质中的cyto.c在2小时释放,caspase-9蛋白表达在4小时达到峰值,caspase-3表达在8小时明显增强。在4小时时,与50 μmol/L RES-BSANP处理的细胞相比,100 μmol/L RES-BSANP处理的细胞中caspase-9表达显著降低(P>0.05)。

结论

RES-BSANP可通过半胱天冬酶依赖性或非依赖性途径诱导SKOV3细胞坏死和凋亡。

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