Induction of apoptosis of ovarian cancer cells and influence on Fas-mediated apoptosis pathway by nanosecond pulsed electric fields.
- Author:
Rumin XIA
1
;
Junying TANG
;
Xue ZHAO
;
Fei GUO
;
Jian WANG
;
Chenguo YAO
Author Information
1. Department of Obstetrics and Gynecology, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
radiation effects;
Caspase 8;
metabolism;
Cell Line, Tumor;
Electromagnetic Fields;
Electroporation;
methods;
Fas Ligand Protein;
Female;
Humans;
Ovarian Neoplasms;
pathology;
fas Receptor;
metabolism
- From:
Journal of Biomedical Engineering
2012;29(6):1144-1149
- CountryChina
- Language:Chinese
-
Abstract:
This paper is to investigate the apoptosis effect of ovarian cancer SKOV3 cells induced by nanosecond plused electric fileds (nsPEFs) and to study its influence on Fas-mediated apoptosis. SKOV3 cell were exposed to the 45kV/cm of field intensity, 30 pulses, and 50ns, 100ns, and 200ns of pulse width, respectively. Flow cytometry were used to assay apoptosis. Agarose gel electrophoresis was used to detect DNA ladder. Real time PCR (RT-PCR) and Western blot analysis were used to measure the expression level of Fas, FasL, caspase-8 and Bid. Flow cytometry results revealed that the late apoptosis rates and (or) necrosis were significantly higher than those in control group (3.03% +/- 0.57%) (P < 0.05), with apoptosis rates and (or) necrosis being (18.31 +/- 0.65%), (45.55% +/- 3.71%), (47.47% +/- 7.01%) in the groups of 50ns, 100ns, 200ns, respectively. A typical DNA ladder pattern of internucleosomal fragmentation was observed in the groups of 50ns and 100ns, but not clear in the 200ns group. RT-PCR results revealed that the mRNA expression of Fas, FasL, caspase8 and Bid were significantly increased in groups of 50ns, 100ns, but significantly decreased in group of 200ns (P < 0.05). Meanwhile, Western blot analysis demonstrated that the Fas, FasL, Caspase-8 and Bid expression were significantly higher in groups of 50ns, 100ns, but significantly lower in group of 200ns (P < 0.05). It indicated that 45kV/cm, 50ns, 100ns nsPEFs could induce apoptosis in ovarian cancer SKOV3 cells and activate Fas-mediated apoptosis pathway.
