Xiao-Yun Yang1, Hai Yu, Wei Lin, Zhi-Lan Peng. 1. Department of Obstetrics and Gynecology, West China Second Hospital, Sichuan University, Chengdu 610041, China.
Abstract
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). CONCLUSION: 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.
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). CONCLUSION: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.