STAT3 silencing with lentivirus inhibits growth and induces apoptosis and differentiation of U251 cells.
Glioblastoma multiforme (GBM), the most common type of central nervous system tumor in humans, is highly proliferative and resistant to apoptosis associated with genetic mutations that deregulate cell cycle. Signal transduction and activation of transcription 3 (Stat3) is a key signal transduction protein that mediated signaling by cytokines and contributed to oncogenesis. It is constitutively activated in numerous cancers including glioblastoma. To determine the effect on proliferation and differentiation of glioblastoma U251 cells after inhibiting STAT3 expression by RNAi, STAT3 gene was silenced with lentivirus vector in U251 cells. We demonstrate that a lentivirus-based shRNA vector had highly infecting efficiency to U251 cells and lentivirus vector-mediated RNAi significantly suppressed Stat3 expression and activation in U251 cells. Knockdown of STAT3 expression by RNAi suppressed the growth and induced apoptosis of U251 cells by down-regulating expression of Bcl-2. It was found that the cell proportion of G0/G1 phase significantly increased after silencing Stat3 by down-regulating expression of cyclin D1. Knockdown of Stat3 also induces morphological changes of U251 cell. It increases significantly expression of myelin basic protein (MBP) in U251 cells. This study demonstrates that STAT3 silencing with lentivirus effectively inhibits STAT3 gene expression and activation. Stat3 is associated with the survival, growth and differentiation of U251 cells. Lentivirus vector-mediated RNAi may be serve as a novel therapeutic strategy for treatment of GBM with expression constitutively and activation of STAT3 gene.
Li
Guang-hui
GH
Third Military Medical University, Chongqing, 400037, China.
Wei
Hong
H
Chen
Zheng-tang
ZT
Lv
Sheng-qing
SQ
Yin
Chang-lin
CL
Wang
Dong-lin
DL
D000704:Analysis of Variance; D017209:Apoptosis / Q000235:genetics*; D002454:Cell Differentiation / Q000235:genetics*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005434:Flow Cytometry / Q000379:methods; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005909:Glioblastoma / Q000503:physiopathology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000502:physiology; D004676:Myelin Basic Protein / Q000378:metabolism; D034622:RNA Interference / Q000502:physiology; D050796:STAT3 Transcription Factor / Q000235:genetics* / Q000378:metabolism*; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods
D005904:Glial Fibrillary Acidic Protein; D004676:Myelin Basic Protein; D050796:STAT3 Transcription Factor; D049452:Green Fluorescent Proteins