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Updated: May 15, 2026
Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence
Published on: November 2, 2014
[Preliminary study of the prohibitin protein and paclitaxel resistance in ovarian cancer]
Juan Tang1, Lanqin Cao, Hong Yi
1Department of Gynecology and Obstetrics, Central South University, Changsha, China.
Objective:
To determine the effect of RNA interference with transferred pshRNA/PHB on the biological characteristics of paclitaxel-resistant ovarian cancer cell lines.
Methods:
Western blot and real time-PCR were used to assay the expression of PHB protein and mRNA in SKOV3/Taxol-25 and SKOV3 cell lines. The SKOV3/Taxol-25 cell lines were transiently transfected by 3 target-specific small hairpin RNA (shRNA) interference fragments with fluorescent protein named the pshRNA427/PHB1, pshRNA248/PHB2, and pshRNA136/PHB3. The empty plasmid transfection via vehicle Lipofectamine2000 served as a negative control. The expression levels of PHB protein and mRNA were detected by Western blot and real time-PCR after the transfection for 48 h. The silence effect of PHB1 and PHB3 groups was obvious. PHB1, PHB3, and the negative control groups were used for the following experiments. MTT and flow cytometry assay were used to test the cell proliferation, IC50 of paclitaxel, and cell apoptosis in the 3 groups.
Results:
The expression levels of PHB protein and mRNA (2(-ΔΔCt)) were significantly higher in SKOV3/Taxol-25 cell line than those in SKOV3 cell line (P<0.05). The expression levels of PHB protein and mRNA were significantly lower in the PHB1 and PHB3 groups than those in the negative control group (P<0.05). The cell proliferations in the PHB1 and PHB3 groups were obviously slower than those in the negative control group after transfection for 48 h and 72 h (P<0.05). The IC50 of paclitaxel in the PHB1 and PHB3 groups significantly decreased after transfection for 72 h compared with the negative control group(P<0.05). The cell apoptotic rate in the PHB1 and PHB3 groups significantly increased after transfection for 48 h compared with the negative control group (P<0.05).
Conclusion:
The shRNA/PHB can effectively suppress the expression of PHB gene in paclitaxel-resistant ovarian cancer cell lines. The cell proliferation in paclitaxel-resistant cell lines with removed PHB gene is significantly reduced. The apoptotic rate and the paclitaxel sensitivity of resistant cell lines with removed PHB gene are significantly increased. PHB gene is related to paclitaxel-resistance and interfering PHB gene expression may reduce paclitaxel resistance in ovarian cancer.
Insights
Silencing the PHB gene using shRNA/PHB significantly reduces proliferation and increases paclitaxel sensitivity in paclitaxel-resistant ovarian cancer cells. This suggests PHB gene interference is a potential strategy to overcome drug resistance in ovarian cancer.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- Paclitaxel resistance is a major challenge in ovarian cancer treatment.
- The PHB gene's role in this resistance is not fully understood.
- Targeting specific genes offers a potential strategy to overcome drug resistance.
Purpose of the Study:
- To investigate the effect of RNA interference targeting the PHB gene on paclitaxel-resistant ovarian cancer cell lines.
- To determine if suppressing PHB gene expression can re-sensitize cells to paclitaxel.
- To analyze the impact on cell proliferation, apoptosis, and drug sensitivity.
Main Methods:
- Utilized Western blot and real-time PCR to measure PHB protein and mRNA expression.
- Transfected paclitaxel-resistant SKOV3/Taxol-25 cells with small hairpin RNA (shRNA) targeting PHB (PHB1, PHB3).
- Assessed cell proliferation (MTT assay), paclitaxel IC50, and apoptosis (flow cytometry) post-transfection.
Main Results:
- PHB gene and protein expression were significantly higher in resistant cells.
- shRNA/PHB effectively reduced PHB expression in transfected cells.
- Suppression of PHB led to decreased cell proliferation, reduced paclitaxel IC50, and increased apoptosis.
Conclusions:
- PHB gene interference effectively suppresses PHB expression in paclitaxel-resistant ovarian cancer cells.
- Reducing PHB gene expression significantly decreases cell proliferation and increases apoptosis.
- Interfering with PHB gene expression enhances paclitaxel sensitivity, indicating its role in drug resistance and potential as a therapeutic target.
