Korean J Physiol Pharmacol.  2025 Mar;29(2):191-204. 10.4196/kjpp.24.132.

Tasquinimod promotes the sensitivity of ovarian cancer cells to cisplatin by down-regulating the HDAC4/p21 pathway

• Li Z
• Wu YH
• Guo YQ
• Min XJ
• Lin Y

• KMID: 2565431
• DOI: http://doi.org/10.4196/kjpp.24.132

Abstract

To investigate whether Tasquinimod can influence cisplatin resistance in drug-resistant ovarian cancer (OC) cell lines by regulating histone deacetylase 4 (HDAC4) or p21, we explored its effects on the cell cycle, and associated mechanisms. RT-PCR and Western blot analyses, flow cytometry, CCK8 assay, and immunofluorescence were utilized to investigate the effects of Tasquinimod on gene expression, cell cycle, apoptosis, viability, and protein levels in OC cells. The results showed that Tasquinimod inhibited cell viability and promoted apoptosis in SKOV3/DDP (cisplatin) and A2780/DDP cells more effectively than DDP alone. In combination with cisplatin, Tasquinimod further enhanced cell apoptosis and reduced cell viability in these cell lines, an effect that could be reversed following HDAC4 overexpression. Tasquinimod treatment down-regulated HDAC4, Bcl-2, and cyclin D1, and CDK4 expression and up-regulated the cleaved-Caspase-3, and p21 expression in SKOV3/DDP and A2780/ DDP cells. Additionally, Tasquinimod inhibited DDP resistance in OC/DDP cells. These effects were similarly observed in OC mouse models treated with Tasquinimod. In conclusion, Tasquinimod can improve OC cells' sensitivity to DDP by down-regulating the HDAC4/p21 axis, offering insights into potential strategies for overcoming cisplatin resistance in OC.

Keyword

Cisplatin; Drug resistance; Drug therapy; Histone deacetylases; Ovarian neoplasms

Figure

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  Fig. 1 Tasquinimod enhanced the anti-proliferative and pro-apoptotic effects of cisplatin in OC/DDP cells. (A) Western blot analysis verified that Tasquinimod inhibited HDAC4 expression in SKOV3/DDP and A2780/DDP cells. (B) CCK8 assays were used to assess the effects of Tasquinimod monotherapy on the viability of SKOV3/DDP and A2780/DDP cells. (C) Flow cytometry was used to detect apoptosis in SKOV3/DDP and A2780/DDP cells treated with Tasquinimod monotherapy. All experiments were independently repeated three times. Tukey’s test and one-way ANOVA were used to analyze statistical significance. Values are presented as mean ± SD. OC, ovarian cancer; HDAC4, histone deacetylase 4; CCK8, cell counting kit-8. *p < 0.05 vs. SKOV3 or A2780 group, #p < 0.05 vs. SKOV3/DDP or A2780/DDP group, &p < 0.05 vs. SKOV3/DDP cells or A2780/DDP cells group, $p < 0.05 vs. SKOV3/DDP cells + cisplatin or A2780/DDP cells + cisplatin group.

• 

  Fig. 2 Tasquinimod promoted apoptosis in OC/DDP cells by inhibiting HDAC4. (A) Western blot analysis of the effect of Tasquinimod on HDAC4 expression. (B) CCK8 assays were used to evaluate the effect of Tasquinimod on cell viability. (C) Flow cytometry was used to detect apoptosis in SKOV3/DDP and A2780/DDP cells treated with Tasquinimod. (D, E) qRT-PCR and Western blot analyses were used to measure the expression levels of Bcl-2 and cleaved-Caspase-3 in tumor cells treated with Tasquinimod. All experiments were independently repeated three times. Tukey’s test and one-way ANOVA were used to analyze statistical significance. Values are presented as mean ± SD. OC, ovarian cancer; HDAC4, histone deacetylase 4; CCK8, cell counting kit-8; NC, negative control. *p < 0.05 vs. SKOV3/DDP + NC or A2780/DDP + NC group, #p < 0.05 vs. SKOV3/DDP or A2780/DDP group, &p < 0.05 vs. SKOV3/DDP + Tasquinimod or A2780/DDP + Tasquinimod group.

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  Fig. 3 Tasquinimod affected the cell cycle of cisplatin-resistant OC/DDP cells. (A) Flow cytometry analysis of the cell cycle in SKOV3/DDP and A2780/DDP cells after Tasquinimod treatment. (B, C) qRT-PCR and Western blot analyses of p21 expression levels in SKOV3/DDP and A2780/DDP cells after Tasquinimod treatment. All experiments were independently repeated three times. Tukey’s test and one-way ANOVA were used to analyze statistical significance. Values are presented as mean ± SD. OC, ovarian cancer; HDAC4, histone deacetylase 4; NC, negative control. *p < 0.05 vs. SKOV3/DDP + NC or A2780/DDP + NC group, #p < 0.05 vs. SKOV3/DDP or A2780/DDP group.

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  Fig. 4 Tasquinimod regulated cell cycle-related gene expression in OC/DDP cells. (A, B) qRT-PCR analysis of p21, cyclin D1, and CDK4 expression levels in SKOV3/DDP cells and A2780/DDP cells after Tasquinimod treatment. (C, D) Immunofluorescence analysis of p21, cyclin D1, and CDK4 expression levels in SKOV3/DDP cells and A2780/DDP cells after Tasquinimod treatment. Scale bars represent 25 μm. (E) CCK8 assays to verify the cell viability in SKOV3/DDP cells and A2780/DDP cells after Tasquinimod treatment. (F) Flow cytometry detection of SKOV3/DDP cell and A2780/DDP cells cycle after Tasquinimod treatment. All experiments were independently repeated six times, Turkey’s test and one-way ANOVA were used to analyze and calculate statistical significance. Values are presented as mean ± SD. OC, ovarian cancer; CCK8, cell counting kit-8; NC, negative control; IOD, integrated optical density. *p < 0.05 vs. SKOV3/DDP or A2780/DDP group, #p < 0.05 vs. SKOV3/DDP + Tas + NC or A2780/DDP + Tas + NC group, &p < 0.05 vs. SKOV3/DDP + Tas or A2780/DDP + Tas group.

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  Fig. 5 Tasquinimod promoted the anti-tumor of cisplatin in OC/DDP cells xenografts in mice. (A–C) Tumor volume and weight were measured and compared in mice after treatment with Tasquinimod combined with cisplatin. (D) Immunofluorescence analysis of HDAC4 expression levels in tumor cells in mice. (E) Western blot analysis of HDAC4, p21, cyclin D1, and CDK4 expression levels in mouse tumor tissues. (F) Flow cytometry analysis of apoptosis in tumor tissues in mice. All experiments were independently repeated three times. Tukey’s test and one-way ANOVA were used to analyze statistical significance. Values are presented as mean ± SD. OC, ovarian cancer; HDAC4, histone deacetylase 4; IOD, integrated optical density. *p < 0.05 vs. SKOV3 or A2780 group, #p < 0.05 vs. SKOV3/DDP or A2780/DDP group.

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