Triptolide induces apoptosis of human acute T lymphocytic leukemia Jurkat cells via inhibiting transcription of human endogenous retrovirus HERV-K Np9 gene
10.3969/j.issn.1673-4254.2015.05.15
- VernacularTitle:雷公藤甲素通过抑制逆转录病毒HERV-K Np9基因转录诱导人急性T淋巴细胞白血病Jurkat细胞凋亡
- Author:
Jianghua CHEN
1
;
Weiwei ZHENG
;
Xudong JIANG
;
Xiaoya LU
;
Rongzhen XU
Author Information
1. 浙江大学医学院附属第二医院血液科
- Keywords:
Triptolide;
acute T lymphocytic leukemia Jurkat cells;
apoptosis;
HERV-K Np9 gene
- From:
Journal of Southern Medical University
2015;(5):702-706
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the molecular mechanisms by which triptolide induces apoptosis of human acute T lymphocytic leukemia Jurkat cells. Methods MTT assay was employed to detect the proliferation inhibition of Jurkat cells by triptolide, and the IC50 was calculated by OriginPro8. Flow cytometry was used to analyze apoptosis of Jurkat cells. Np9 mRNA levels were detected by RT-PCR and analyzed quantitatively by Kodak 1D 3.6 software. Correlation between the inhibition of Np9 transcription and the cell apoptosis was analyzed by SPSS 19.0.Western blotting was employed to determine Np9 downstream signaling molecules c-myc,β-catenin, ERK, AKT and Notch1 protein level in Jurkat cells after exposure to different concentrations of triptolide for 48h. Results Triptolide treatment resulted in dose-dependent inhibition of Jurkat cells proliferation and its IC50 was 12.7nmol/L. Triptolide induced apoptosis of Jurkat cells in dose-dependent manner. Furthermore, triptolide inhibited Np9 mRNA transcription level in Jurakt cells in a dose-dependent manner. There was a correlation between the triptolide-mediated the apoptosis and the inhibition of Np9 transcription of Jurkat cells (R2=0.907). Western blotting results displayed that triptolide inhibited transcription levels of Np9 mRNA with a concomitant decrease of its downstream signaling molecules c-myc,β-catenin, ERK, AKT and Notch1 at protein levels. Conclusion Inhibition of HERV-K Np9 mRNA and its downstream signaling molecules c-myc, β-catenin, ERK, Akt and Notch1 protein might be one of important molecular mechanisms by which triptolide induces apoptosis of human acute T lymphocytic leukemia Jurkat cells.
