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[Effects of RNA interference silencing RhoC gene upon paclitaxel sensitivity in ovarian cancer cell lines]
Ying Pan1, Wen-ying Zhang, Ji-yan Zou
1Department of Gynecology and Obstetrics, China Japan Union Hospital of Jilin University, Changchun 130033, China.
Objective:
To explore the changes of paclitaxel sensitivity by RNA interference of RhoC gene in ovarian cancer cell lines.
Methods:
The microRNA (miRNA) targeting RhoC gene was designed and synthesized by in vitro transcription and transfected into ovarian cancer cell line SKOV3. The mRNA and protein of RhoC were detected by the means of RT-PCR and Western blot. The changes of paclitaxel sensitivity after interference were examined by methyl thiazolyl tetrazolium (MTf) assay. At the same time, the mRNA of apoptosis gene Caspase-3 and survivin were evaluated by RT-PCR.
Results:
In SKOV3 cell, the mRNA and protein of RhoC dramatically decreased at 24, 48 and 72 hours post-transfection. The sensitivity of SKOV3 cell line to paclitaxel increased significantly after turning off the RhoC gene. The proapoptotic gene Caspase-3 increased significantly and anti-apoptosis gene survivin dramatically decreased after the combination treatment of paclitaxel and RhoC-miRNA in comparison with paclitaxel alone treatment.
Conclusion:
The sensitivity of ovarian cancer cell lines SKOV3 to paclitaxel can be enhanced by RNA interfering RhoC gene. The synergistic mechanism is possibly correlated with the up-regulation of Caspase-3 gene and the down-regulation of survivin gene.
Insights
RNA interference targeting the RhoC gene enhances paclitaxel sensitivity in ovarian cancer cells. This approach increases the pro-apoptotic Caspase-3 gene and decreases the anti-apoptotic survivin gene, offering a potential synergistic treatment strategy.
Area of Science:
- Molecular Biology
- Oncology
- Gene Regulation
Background:
- Ovarian cancer remains a significant health challenge with limited treatment options.
- Paclitaxel is a key chemotherapeutic agent, but resistance can develop.
- RhoC gene expression is implicated in cancer progression and drug resistance.
Purpose of the Study:
- To investigate the effect of RhoC gene silencing on paclitaxel sensitivity in ovarian cancer cells.
- To explore the underlying molecular mechanisms involving apoptosis-related genes.
Main Methods:
- Ovarian cancer cell line SKOV3 was transfected with microRNA targeting RhoC.
- RhoC mRNA and protein levels were assessed using RT-PCR and Western blot.
- Paclitaxel sensitivity was evaluated via MTT assay, alongside Caspase-3 and survivin gene expression analysis.
Main Results:
- RhoC gene silencing significantly reduced RhoC mRNA and protein expression in SKOV3 cells.
- Paclitaxel sensitivity of SKOV3 cells increased markedly following RhoC interference.
- Caspase-3 (pro-apoptotic) gene expression was upregulated, while survivin (anti-apoptotic) gene expression was downregulated.
Conclusions:
- RNA interference of the RhoC gene enhances paclitaxel efficacy in ovarian cancer cell line SKOV3.
- The observed synergy is potentially mediated by the modulation of Caspase-3 and survivin gene expression.
- Targeting RhoC offers a promising strategy to overcome paclitaxel resistance in ovarian cancer.
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