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[Study on the Relationship Between XIAP Gene and Resistance of Taxol in Ovarian Cancer]
Chi Yue1,2, Ran-Hong Li3, Chen Chen1,2
1Department of Obstetrics and Gynecology,West China Second University Hospital,Sichuan University,Chengdu 610041,China.
Objective:
To research the expression of X-linked inhibitor of apoptosis protein gene (XIAP) on paclitaxel resistance in ovarian cancer.
Methods:
A2780 and A2780/T cells were treated with paclitaxel respectively at the concentrations of 5 ng/mL,10 ng/mL,20 ng/mL , 40 ng/mL,80 ng/mL,160 ng/mL,320 ng/mL,then the inhibition rate of cells were detected by MTT assay. The expression of XIAP mRNA and protein among the A2780 and A2780/T cells treated respectively with paclitaxel at the concentration of 100 ng/mL was detected by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and Western blot. The A2780/T cells were divided into blank group,empty group,small interfering RNA (siRNA) XIAP group and siRNA-non-specific group. The expression of XIAP mRNA and protein of four groups were detected by RT-qPCR and Western blot. Apoptotic rate of these groups with addition of paclitaxel at the concentrations of 0 ng/mL,1 000 ng/mL,1 500 ng/mL,2 000 ng/mL and 2 500 ng/mL were detected by flow cytometry.
Results:
After the treatments on A2780 and A2780/T cells with the different concentrations of paclitaxel,the inhibition rate of A2780 cells were gradually increased with the increased paclitaxel concentrations (P<0.05),while there were no obvious differences in A2780/T cells (P>0.05). After the treatment on these cells with paclitaxel at the concentration of 100 ng/mL,the expression of XIAP mRNA was lower than that non-treatment with paclitaxelin A2780 cells (P<0.05),and the expression of XIAP mRNA in the A2780/T cells were no statistical significance between the treatment group and non-treatment group with paclitaxel (P>0.05). However,the expression of A2780/T cells'XIAP mRNA and protein treated with paclitaxel were higher than A2780 cells' (P<0.05). The expression of XIAP mRNA and protein in siRNA-XIAP group was lower than those of other groups (P<0.05). The apoptotic rate of siRNA-XIAP group was higher than those of other groups treated with the paclitaxel at concentrations of 2 000 ng/mL and 2 500 ng/mL (P<0.05).
Conclusion:
XIAP's high expression on mRNA and protein was correlated with ovarian cancer paclitaxel-resistance,specific siRNA can promote cell apoptosis by reducing the expression of XIAP,and increase the sensitivity of drug-resistant cancer cells to paclitaxel.
Insights
High X-linked inhibitor of apoptosis protein gene (XIAP) expression correlates with paclitaxel resistance in ovarian cancer. Silencing XIAP with siRNA enhances apoptosis and resensitizes resistant cells to paclitaxel.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Ovarian cancer remains a significant health concern, with paclitaxel resistance posing a major therapeutic challenge.
- The X-linked inhibitor of apoptosis protein gene (XIAP) is implicated in apoptosis regulation and cancer progression.
Purpose of the Study:
- To investigate the role of XIAP expression in paclitaxel resistance in ovarian cancer.
- To evaluate the potential of targeting XIAP to overcome drug resistance.
Main Methods:
- Cell lines (A2780 and paclitaxel-resistant A2780/T) were treated with varying paclitaxel concentrations.
- XIAP mRNA and protein levels were assessed using RT-qPCR and Western blot.
- Small interfering RNA (siRNA) targeting XIAP was used to modulate its expression, followed by apoptosis assays (flow cytometry).
Main Results:
- Paclitaxel resistance in A2780/T cells was associated with higher XIAP mRNA and protein expression compared to sensitive A2780 cells.
- siRNA-mediated knockdown of XIAP significantly reduced XIAP levels.
- XIAP knockdown increased apoptosis rates and enhanced sensitivity to paclitaxel in resistant cells.
Conclusions:
- Elevated XIAP expression is linked to paclitaxel resistance in ovarian cancer.
- Targeting XIAP with siRNA can overcome paclitaxel resistance by promoting apoptosis and increasing drug sensitivity.
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