Inhibitory effects of c-erbB-2 antisense oligonucleotide transfection on uterine endometrial cancer Ishikawa cell lines
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1Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shen Yang, PR China. zhaofujie63@yahoo.com.cn
Background:
C-erbB-2 is a chief proto-oncogene of endometrial cancer, which plays an important role in the biological behavior of endometrial cancer. Its overespression is an important factor of poor progrosis. The objective of this study was to investigate the treatment effects of transfecting c-erbB-2 antisense oligonucleotide (ASODN) on the uterine endometrial cancer Ishikawa cell line.
Materials And Methods:
The c-erbB-2 expression on Ishikawa cell membranes was determined by immunohistochemistry and then the aim was to transfect ASODN into Ishikawa cells and assay the cellular growth inhibition by MTT, to observe the cellular ultrastructure changes under transmission electron microscope (TEM), and to assay the cellular apoptotic rate, c-erbB-2 mRNA and protein expression by flow cytometry, RT-PCR and Western blot, respectively.
Results:
C-erbB-2 protein was positively expressed on Ishikawa cell membranes. MTT showed that when the concentration of transfecting ASODNs was 0.3 uM and 0.6 uM, cell growth inhibition rates were 55.43% and 76.12%, respectively. After transfecting 0.3 uM ASODN, the Ishikawa cell ultrastructure was obviously damaged, the cellular apoptotic rate was 72.21%, and the c-erbB-2 mRNA and protein expression were 45.71% and 34.52%, respectively, compared with those of the normal control cells.
Conclusions:
Transfecting c-erbB-2 ASODN can obviously suppress its mRNA and protein expression in Ishikawa cells, cause cellular apoptosis and inhibit cell growth. It may have an important role in the gene therapy of endometrial cancer.
Insights
Transfecting c-erbB-2 antisense oligonucleotide (ASODN) into endometrial cancer cells significantly inhibits cell growth and suppresses c-erbB-2 expression. This approach shows promise for gene therapy in endometrial cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- C-erbB-2 is a proto-oncogene crucial to endometrial cancer development and progression.
- Overexpression of C-erbB-2 is linked to poor prognosis in endometrial cancer patients.
Purpose of the Study:
- To investigate the therapeutic effects of transfecting c-erbB-2 antisense oligonucleotide (ASODN) on the Ishikawa endometrial cancer cell line.
- To assess the impact of ASODN on cell growth, apoptosis, and c-erbB-2 expression.
Main Methods:
- Immunohistochemistry to detect c-erbB-2 expression.
- MTT assay for cell growth inhibition.
- Transmission electron microscopy (TEM) for ultrastructural changes.
- Flow cytometry, RT-PCR, and Western blot to analyze apoptosis, mRNA, and protein expression.
Main Results:
- Transfection with 0.3 uM and 0.6 uM ASODN resulted in 55.43% and 76.12% cell growth inhibition, respectively.
- At 0.3 uM ASODN, significant ultrastructural damage, a 72.21% apoptotic rate, and reduced c-erbB-2 mRNA (45.71%) and protein (34.52%) expression were observed.
Conclusions:
- Transfecting c-erbB-2 ASODN effectively suppresses c-erbB-2 mRNA and protein levels in Ishikawa cells.
- ASODN treatment induces cellular apoptosis and inhibits endometrial cancer cell growth.
- This strategy holds potential for endometrial cancer gene therapy.
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