[Effects of ARLTS1 gene on growth and apoptosis of epithelial ovarian cancer SKOV3 cells].
OBJECTIVE
To investigate the effect of ARLTS1 (ADP-ribosylation factor-like tumor suppressor gene 1) on the growth and apoptosis of ovarian cystadenocarcinoma cell line SKOV3.


METHODS
The recombinant plasmid pCMV-Tag 3B-ARLTS1 was constructed and transfected into SKOV3 cells by liposome protocol. The expression of ARLTS1 mRNA and its protein were examined by RT-PCR and Western blotting. Effects of ARLTS1 gene on growth and apoptosis of the transfected cells were evaluated by MTT assay and flow cytometry (FCM). The alterations of caspase-3 and bcl-2 protein levels were examined by Western blotting.


RESULTS
The stable transfection of pCMV-Tag 3B-ARLTS1 in SKOV3 cells were obtained and verified by RT-PCR and Western blotting methods. After transfected with ARLTS1, more SKOV3 cells were observed in S phase by FCM compared with those in the other two groups (P is 0.035 and 0.011, respectively). The 36.7% apoptotic index of ARLTS1 transfected cell was significantly higher than that of the other two groups (P < 0.001). The growth of ARLTS1 transfected cells was dramatically inhibited compared with cells transfected with Vector (P < 0.05). Western blotting indicated a significantly decrease in caspase-3 and bcl-2 protein levels in cells transfected with ARLTS1 compared with SKOV3 cells (P is 0.021 and 0.013, respectively).


CONCLUSIONS
ARLTS1 transfection can inhibit proliferation and induce apoptosis of SKOV3 cells in vitro and down-regulate the expression of caspase-3 and bcl-2. ARLTS1 gene may be a candidate tumor suppressor gene in ovarian cancer.
Yang
Xiao-Yun
XY
Department of Obstetrics and Gynecology, West China Second Hospital, Sichuan University, Chengdu 610041, China.
Yu
Hai
H
Lin
Wei
W
Peng
Zhi-Lan
ZL
D020727:ADP-Ribosylation Factors / Q000235:genetics* / Q000378:metabolism; D017209:Apoptosis / Q000235:genetics*; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003536:Cystadenocarcinoma / Q000235:genetics / Q000473:pathology; D005260:Female; D006801:Humans; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000473:pathology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D053148:Caspase 3; D020727:ADP-Ribosylation Factors; C500039:ARL11 protein, human