TAZ Regulates the Cisplatin Resistance of Epithelial Ovarian Cancer Cells via the ANGPTL4/SOX2 Axis

Figures

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  Figure 1. TAZ was highly expressed in drug-resistant EOC cells. The A2780/DDP cell models were established. (a) Cell viability was detected by CCK-8. (b) After EOC cells were treated with cisplatin at different concentrations for 48 hours, the drug sensitivity of EOC cells was detected by CCK-8. (c) TAZ mRNA expression was determined using RT-qPCR. (d) Protein expression of TAZ was measured via WB. Cell experiment was repeated three times. Data were expressed as mean ± SD, and independent sample t test was used for comparisons between the two groups. ∗∗P < 0.01.

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  Figure 2. Knockdown of TAZ improved the drug sensitivity of EOC cells. Drug-resistant EOC cells and EOC cells were transfected with siR-TAZ and pcDNA3.1-TAZ, respectively. (a) RT-qPCR was used to detect the expression of TAZ. (b) WB was employed to determine the protein expression of TAZ. (c) After cells were treated with different concentrations of cisplatin for 48 hours based on transfection with siR-TAZ and pcDNA3.1-TAZ, CCK-8 was employed to measure the drug sensitivity of EOC cells. Cell experiment was repeated three times. Data were shown as mean ± SD, and one-way ANOVA was adopted for comparisons among groups. Tukey's multiple comparisons test was implemented for the post hoc test. ∗P < 0.05, ∗∗P < 0.01.

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  Figure 3. Knockdown of TAZ inhibited the drug-resistant EOC cell stemness. The drug-resistant EOC cells were transfected with siR-TAZ or treated with 120 nM cisplatin for 48 hours. (a) Apoptosis rate was measured by flow cytometry. (b) The expression of biomarkers for CSCs was detected by WB. (c) Impairment of tumorsphere formation was observed by spheroid formation assay. Cell experiment was repeated three times. Data were presented as mean ± SD, and one-way ANOVA was used for comparisons among groups. Tukey's multiple comparisons test was employed for the post hoc test. ∗∗P < 0.01.

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  Figure 4. ANGPTL4 was a direct target gene of TAZ to regulate cell resistance. (a) ChIP-qPCR was used to verify the binding relationship of TAZ and ANGPTL4. (b) WB was adopted to detect the protein expression of ANGPTL4. (c) ELISA was used to determine the expression of ANGPTL4 in cell supernatant. (d) CCK-8 was employed to detect the drug sensitivity of drug-resistant EOC cells. (e) Flow cytometry was implemented to measure the apoptosis rate. Cell experiment was repeated three times. Data were expressed as mean ± SD, and one-way ANOVA analysis was employed for comparisons among groups. Tukey's multiple comparisons test was used for the post hoc test. ∗P < 0.05, ∗∗P < 0.01.

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  Figure 5. ANGPTL4 regulated the DDP sensitivity of drug-resistant EOC cells through SOX2. The drug-resistant EOC cells were transfected with siR-SOX2 based on TAZ knockdown and ANGPTL4 overexpression. (a) Protein expression of SOX2 was detected by WB. (b) Drug sensitivity was measured by CCK-8. (c) Apoptosis rate was determined by flow cytometry. Cell experiment was repeated three times. Data are presented as mean ± SD and one-way ANOVA was used for comparisons among groups. Tukey's multiple comparisons test was adopted for the post hoc test. ∗P < 0.05, ∗∗P < 0.01.