Inducing Apoptosis of Human Nasopharyngeal Carcinoma Cell Line CNE-2Z by bcl-xL Short hairpin RNA
10.3321/j.issn:1000-467X.2005.06.002
- VernacularTitle:bcl-xL短发夹状RNA诱导人鼻咽癌细胞株CNE-2Z凋亡
- Author:
Cheng-Wei HE
1
;
Fang LIU
;
Yue-Fei ZHANG
;
Tong LIANG
;
Ke-Yuan ZHOU
Author Information
1. 广东医学院
- Keywords:
Nasopharyngeal neoplasms;
Short hairpin RNA;
bcl-xL;
Apoptosis
- From:Chinese Journal of Cancer
2005;24(6):646-652
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND & OBJECTIVE: Recent studies showed overexpression of bcl-xL in human nasopharyngeal carcinoma (NPC) cell line CNE-2Z; it may play a pivotal role in tumorigenesis, metastasis, and drug resistance of NPC. This study was to explore inducing effect of bcl-xL short hairpin RNA (shRNA) on apoptosis of CNE-2Z cells. METHODS: After transfection of recombinant plasmid pmU6-RNAi expressing bcl-xL shRNA,apoptotic CNE-2Z cells were detected by fluorescent staining and flow cytometry (FCM). mRNA levels of bcl-xL, bcl-2, survivin, and caspase-3 was detected by reverse transcription-polymerase chain reaction (RT-PCR); while protein levels of Bcl-xL, Caspase-3, and P53 were detected by Western blot.RESULTS: When treated with pmU6-RNAi for 24 h, an obvious apoptotic peak of CNE-2Z cells appeared; cell shrinkage, chromatin condensation, and nuclear fragmentation were observed in most cells under fluorescent microscope. RT-PCR analysis showed that pmU6-RNAi down-regulated mRNA levels of bcl-xL, bcl-2, and caspase-3, but had little or no effect on mRNA level of survivin; Western blot analysis showed an obvious reduction in protein levels of Bcl-xL and Caspase-3, and a great increase in protein level of P53.CONCLUSIONS: bcl-xL shRNA can induce apoptosis of CNE-2Z cells, which may be closely related to down-regulation of bcl-2, caspase-3 and p53. bclxL shRNA may be helpful for developing gene therapy for NPC.
