Effect of silencing NOTCH1 gene by shRNA interference on AKT/mTOR pathway in mantle cell lymphoma.
10.7534/j.issn.1009-2137.2014.06.021
- Author:
Yi-Qun HUANG
1
;
Xiao-Lu HUANG
1
;
Xu-Dong MA
2
Author Information
1. Department of Hematology, Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou 363000, Fujian Province, China.
2. Department of Hematology, Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou 363000, Fujian Province, China. E-mail: maxudong005@hotmail.com.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Caspase 3;
Caspase 9;
Cell Line, Tumor;
Cell Proliferation;
Humans;
Lymphoma, Mantle-Cell;
genetics;
Proto-Oncogene Proteins c-akt;
RNA, Messenger;
RNA, Small Interfering;
genetics;
Receptor, Notch1;
genetics;
Signal Transduction;
TOR Serine-Threonine Kinases;
Transfection
- From:
Journal of Experimental Hematology
2014;22(6):1616-1620
- CountryChina
- Language:Chinese
-
Abstract:
The study was purposed to investigate the effect of silencing NOTCH1 gene by shRNA interference on the proliferation, apoptosis and the expression of AKT signaling pathway-related proteins in mantle cell lymphoma Jeko-1 cell line. The hairpin-like oligonucleotide sequences targeting NOTCH1 gene were designed and transfected into Jeko-1 cells by lipofectamine (TM) 2 000. NOTCH1 mRNA and protein were detected respectively by RT-PCR and Western blot. Cell growth was determined by MTT. Cell apoptosis was analyzed by flow cytometry. The expressions of BCL-2, BAX, procaspase-3, procaspase-9, Akt, p-Akt, p-mTOR, p-P70S6K were detected by Western blot. The results showed that NOTCH1 mRNA expression was markedly suppressed by the shRNA targeting NOTCH1. NOTCH1 shRNA suppressed the proliferation of Jeko-1 cells and induced apoptosis of these cells. The cell apoptotic rate was (34.5 ± 3.4)%, (2.4 ± 1.3) %, (1.7 ± 0.6) % in NOTCH1 shRNA, Neg-shRNA and blank groups, respectively, and the difference between them was statistically significant (P < 0.01). NOTCH1 shRNA down-regulated the expression of BCL-2, procaspase-3, procaspase 9, p-Akt, p-mTOR and p-70S6K, up-regulated the expression of BAX, but no change protein expression of Akt was observed. It is concluded that the silencing NOTCH1 gene expression by shRNA interference may inhibit Jeko-1 proliferation, induce the cell apoptosis, and the mechanisms may be associated with the inhibition of Akt/mTOR signaling pathway by dephosphorylation.
