Effects of Sam68 gene silence on proliferation of acute T lymphoblastic leukemia cell line Jurkat.
10.7534/j.issn.1009-2137.2014.04.003
- Author:
Chi-Juan WANG
1
;
Hua XU
1
;
Hai-Rui ZHANG
1
;
Jian WANG
1
;
Ya-Ni LIN
1
;
Tian-Xiang PANG
1
;
Qing-Hua LI
2
Author Information
1. State key Laboratory of Experimental Hematology, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China.
2. State key Laboratory of Experimental Hematology, Institute of Hematology & Hospital of Blood Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China. E-mail: qhli@ihcams.ac.cn.
- Publication Type:Journal Article
- MeSH:
Adaptor Proteins, Signal Transducing;
genetics;
Cell Proliferation;
DNA-Binding Proteins;
genetics;
Genetic Vectors;
Humans;
Jurkat Cells;
Lentivirus;
genetics;
Precursor Cell Lymphoblastic Leukemia-Lymphoma;
genetics;
RNA Interference;
RNA, Messenger;
genetics;
RNA, Small Interfering;
genetics;
RNA-Binding Proteins;
genetics
- From:
Journal of Experimental Hematology
2014;22(4):894-898
- CountryChina
- Language:Chinese
-
Abstract:
This study was purpose to investigate the effect of Sam68 gene silence on proliferation of human acute T lymphoblastic leukemia cell line Jurkat. The sequence of shRNA targeting the site 531-552 of Sam68 mRNA was designed and chemically synthesized, then a single-vector lentiviral, Tet-inducible shRNA-Sam68 system (pLKO-Tet-On) was constructed; next the Jurkat cells were infected with lentivirus to create stable cell clones with regulatable Sam68 gene expression. The inhibitory efficiency of Sam68 gene was assayed by Real-time PCR and Western blot; the cell activity of Jurkat cells was detected with MTT assay; the change of colony forming potential of Jurkat cells was analyzed by colony forming test; the cell cycle distribution was tested by flow cytometry. The results indicated that the expression of Sam68 in experimental cells was statistically decreased as compared with that of the control cells; the cells activity and colony forming capacity of the Jurkat cells with Sam68 gene silence were significantly inhibited; with Sam68 gene silencing, the percentage of S phase cells was significantly increased, while the percentage of G2 phase cells was significantly decreased. It is concluded that the silencing Sam68 gene using shRNA interference can effectively inhibit the proliferation of human acute T lymphoblastic leukemia cell line Jurkat.
