[Effects of vesicular stomatitis virus matrix protein on proliferation and apoptosis of mouse LL/2c cells].
OBJECTIVE
Vesicular stomatitis virus (VSV), an oncolytic virus, is an attractive candidate for tumor therapy. Although previous studies showed obvious antitumor effects of VSV on human A549 and mouse LL/2c tumor models, the clinical application of a live virus is confronted with the problem of bio-safety. The matrix (M) protein of VSV is related to the antitumor effect of VSV. This study was to investigate the effects of VSV-M protein on the proliferation and apoptosis of mouse LL/2c tumor cells.


METHODS
A eukaryotic expression plasmid pcDNA3.1-M encoding VSV-M protein was constructed by molecular cloning technique, and analyzed by enzyme digestion, polymerase chain reaction (PCR), and DNA sequencing, then transfected into LL/2c cells. The expression of VSV-M protein in LL/2c cells was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The effect of VSV-M protein on proliferation of LL/2c cells was assessed by MTT assay; its effect on cell apoptosis was assessed by DNA ladder and Hoechst 33528 staining.


RESULTS
A plasmid pcDNA3.1-M encoding VSV-M protein was constructed successfully and identified. After transfection, the expression of VSV-M protein was detected in LL/2c cells, morphologic changes of LL/2c cells was observed under microscope, inhibition rate of cell survival was 41.3% (P<0.05), DNA ladder was detected, apoptotic nuclei was observed.


CONCLUSIONS
VSV-M transfection could inhibit proliferation and induce apoptosis of LL/2c cells.
Zhao
Ju-Mei
JM
State Key Laboratory of Biotherapy, Cancer Center, West China Hospital, Sichuan University, Chengdu, Sichuan, PR China.
Liu
Tao
T
Xu
Jian-Rong
JR
Wang
Guo-Qing
GQ
Du
Xiao-Bo
XB
Wen
Yan-Jun
YJ
D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism; D017209:Apoptosis; D018827:Carcinoma, Lewis Lung / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018076:DNA, Complementary / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D005822:Genetic Vectors; D051379:Mice; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism*
D000970:Antineoplastic Agents; D018076:DNA, Complementary; D004273:DNA, Neoplasm; C094460:M protein, Vesicular stomatitis virus; D011994:Recombinant Proteins; D014763:Viral Matrix Proteins