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[Growth inhibitory effects of lipofectamine-mediated DCC gene on ovarian epithelial carcinoma]
Pei-ling Li1, Chun-jie Hu, Chang-min Li
1Department of Obstetrics and Gynecology, Second Affiliated Hospital of Harbin Medical University, Harbin 150086, China.
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.
Insights
The deleted colorectal carcinoma (DCC) gene inhibits ovarian cancer cell growth. Introducing DCC into SKOV3 cells significantly reduced proliferation and altered cell cycle, indicating its role in ovarian cancer development.
Area of Science:
- Molecular Biology
- Genetics
- Oncology
Background:
- Ovarian cancer is a significant health concern with limited treatment options.
- The deleted colorectal carcinoma (DCC) gene's role in ovarian cancer is not fully understood.
Purpose of the Study:
- To investigate the inhibitory effects of the DCC gene on the ovarian cancer cell line SKOV3.
- To determine if lipofectamine-mediated DCC gene transfer can suppress ovarian cancer cell growth.
Main Methods:
- Constructed a recombinant eukaryotic expression vector (pcDNA3.1(+)-DCC) with human DCC cDNA.
- Transfected SKOV3 cells with the vector using lipofectamine, creating SKOV3/DCC, SKOV3/Neo, and SKOV3 cell lines.
- Analyzed DCC mRNA and protein expression using RT-PCR and immunocytochemistry.
Main Results:
- Successfully achieved stable expression of exogenous DCC in SKOV3 cells.
- SKOV3/DCC cells exhibited significantly slower growth rates compared to control groups (SKOV3/Neo and SKOV3).
- Cell cycle analysis showed an increase in G1 phase and a decrease in S phase in SKOV3/DCC cells, indicating growth retardation.
Conclusions:
- The DCC gene plays a crucial role in the development of ovarian carcinomas.
- Gene therapy targeting DCC may offer a potential therapeutic strategy for ovarian cancer.
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