Zhonghua fu chan ke za zhi · 2006
[Growth inhibitory effects of lipofectamine-mediated DCC gene on ovarian epithelial carcinoma].
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Li Pei-ling, Hu Chun-jie, Li Chang-min, Meng Chun-yan, Gao Lei
Аннотация
OBJECTIVE: To study the inhibitory effects of lipofectamine-mediated deleted colorectal carcinoma gene on ovarian epithelial carcinoma (ovarian cancer) cell line SKOV3.
METHODS: We constructed a recombinant eukaryotic expression vector pcDNA3.1 (+)-DCC containing exogenous human DCC cDNA and vector with neomycin resistance gene, which were introduced by lipofectamine-mediated gene transfection into SKOV3 cell line that does not express DCC endogenously, thus forming SKOV3/DCC. Therefore, the experimental cells were classified into SKOV3/DCC, SKOV3/Neo and SKOV3. By using reverse transcriptase-polymerase chain reaction and immunocytochemistry, the expression of DCC mRNA and its protein were examined.
RESULTS: Exogenous DCC had successfully been transferred into SKOV3 cells and obtained permanent expression. The growth speed of SKOV3/DCC was slower than the other two groups, there was significant difference between them (P < 0.01). SKOV3/DCC clones number was 38 +/- 8, while SKOV3 and SKOV3/Neo were 192 +/- 8 and 186 +/- 10, respectively, there was significant difference between them (P < 0.01). The percentage of G(1) phase cells increased to 78.0%, which that of S phase decreased to 5.3% by analyzing cell cycle, there was significant difference between them (20.0% and 3.2%, P < 0.01). The ultrastructural changes of the cells were observed under electron microscope, revealing growth retardation.
CONCLUSION: DCC gene played an important role in generation and development of ovarian carcinomas.
METHODS: We constructed a recombinant eukaryotic expression vector pcDNA3.1 (+)-DCC containing exogenous human DCC cDNA and vector with neomycin resistance gene, which were introduced by lipofectamine-mediated gene transfection into SKOV3 cell line that does not express DCC endogenously, thus forming SKOV3/DCC. Therefore, the experimental cells were classified into SKOV3/DCC, SKOV3/Neo and SKOV3. By using reverse transcriptase-polymerase chain reaction and immunocytochemistry, the expression of DCC mRNA and its protein were examined.
RESULTS: Exogenous DCC had successfully been transferred into SKOV3 cells and obtained permanent expression. The growth speed of SKOV3/DCC was slower than the other two groups, there was significant difference between them (P < 0.01). SKOV3/DCC clones number was 38 +/- 8, while SKOV3 and SKOV3/Neo were 192 +/- 8 and 186 +/- 10, respectively, there was significant difference between them (P < 0.01). The percentage of G(1) phase cells increased to 78.0%, which that of S phase decreased to 5.3% by analyzing cell cycle, there was significant difference between them (20.0% and 3.2%, P < 0.01). The ultrastructural changes of the cells were observed under electron microscope, revealing growth retardation.
CONCLUSION: DCC gene played an important role in generation and development of ovarian carcinomas.