The impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy
Objective To investigate the impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy (NACT). Methods Tumor biopsy tissue samples were collected before NACT from 47 patients with advanced high-grade serous ovarian cancer (HGSOC) who underwent NACT and intermediate tum...
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Editorial Office of Journal of Precision Medicine
2025-08-01
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| Series: | 精准医学杂志 |
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| Online Access: | https://jpmed.qdu.edu.cn/fileup/2096-529X/PDF/1754471601716-1632270267.pdf |
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| author | LIU Hui, CHEN Yunqing, QI Huiyang, SUN Xin, LOU Yanhui |
| author_facet | LIU Hui, CHEN Yunqing, QI Huiyang, SUN Xin, LOU Yanhui |
| author_sort | LIU Hui, CHEN Yunqing, QI Huiyang, SUN Xin, LOU Yanhui |
| collection | DOAJ |
| description | Objective To investigate the impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy (NACT). Methods Tumor biopsy tissue samples were collected before NACT from 47 patients with advanced high-grade serous ovarian cancer (HGSOC) who underwent NACT and intermediate tumor cell reduction surgery in Department of Gynecology, The Affiliated Hospital of Qingdao University, from June 2017 to June 2019. Immunohistochemistry was used to measure the expression level of RAD51 protein in tumor tissue, and the correlation between its expression level and the sensitivity to NACT was analyzed. SKOV3 cells were divided into groups a-f, and cisplatin (DDP)-resistant SKOV3 cells were divided into groups A-F; the cells in groups b-f or groups B-F were transfected with NC-siRNA, siRNA-RAD51-264, siRNA-RAD51-416, siRNA-RAD51-608, and siRNA-RAD51-807, respectively, and then quantitative real-time PCR was used to measure the mRNA expression levels of RAD51 in each group of SKOV3 cells and DDP-resistant SKOV3 cells. The DDP-resistant SKOV3 cells were divided into SKOV3-Control group (control group), SKOV3-DDP-Control group (DDP control group), SKOV3-DDP-NC-siRNA group (transfected with NC-siRNA), and SKOV3-DDP-siRNA-RAD51 group (transfected with siRNA-RAD51), and Western blotting was used to measure the expression level of RAD51 protein in each group of cells. The DDP-resistant SKOV3 cells were divided into NC-siRNA group (transfected with NC-siRNA) and siRNA-RAD51 group (transfected with siRNA-RAD51-264), and DDP at various concentrations (0.39, 0.78, 1.56, 3.12, 6.25, 12.50, and 25.00 mg/L) was added for culture for 48 h; CCK-8 assay was used to measure the viability of DDP-resistant SKOV3 cells after transfection. The DDP-resistant SKOV3 cells were divided into Control group, NC-siRNA group (transfected with NC-siRNA), siRNA-RAD51 group (transfected with siRNA-RAD51), and siRNA-RAD51+DDP group (transfected with siRNA-RAD51 and subsequently treated with DDP), and flow cytometry was used to observe cell cycle alterations and apoptosis rate in DDP-resistant SKOV3 cells. Results Immunohistochemistry showed that there were no significant differences in age, FIGO stage (stage Ⅲ or Ⅳ), lymph node metastasis, ascites, and initial serum CA125 level between the patients with high RAD51 expression and those with low expression (P>0.05), and there was a significant difference in the expression level of RAD51 protein in tumor tissue between the chemotherapy-sensitive group and the insensitive group (χ2=10.85,P<0.05). RT-qPCR showed that the relative mRNA expression level of RAD51 in SKOV3-DDP cells was significantly higher than that in SKOV3 cells (t=6.73,P<0.05), and the mRNA expression level of RAD51 in group c/C was significantly lower than that in group a/A (F=7.81,6.58,P<0.05); there was no significant difference in the mRNA expression level of RAD51 between groups d/D, e/E, f/F and group a/A (P>0.05). Western blotting showed that compared with group A, group B had a significant increase in the protein expression level of RAD51 in cells, and compared with groups A and B, group D had a significant reduction in the protein expression level of RAD51 (F=10.56,P<0.05). CCK-8 assay showed that compared with the NC-siRNA group, the siRNA-RAD51-264 group had a significant reduction in cell viability at DDP concentrations of 3.12, 6.25, 12.50, and 25.00 mg/L (t=3.72-63.90,P<0.05). Flow cytometry showed that compared with the Control group and the NC-siRNA group, the siRNA-RAD51 group and the siRNA-RAD51+DDP group had a significant increase in the proportion of cells in the G0/G1 phase, and compared with the siRNA-RAD51 group, the siRNA-RAD51+DDP group had a significant increase in the proportion of cells in the G0/G1 phase (F=23.29,P<0.05); compared with the Control group and the NC-siRNA group, the siRNA-RAD51+DDP group had a significant reduction in the proportion of cells in the S phase (F=12.95,P<0.05). Compared with the Control group and the NC-siRNA group, both the siRNA-RAD51 group and the siRNA-RAD51+DDP group had a significant increase in apoptosis rate (F=16.34,P<0.05). Conclusion Patients with a high expression level of RAD51 protein in ovarian cancer tissue tend to have poorer sensitivity to NACT, and knocking down the expression of RAD51 gene in DDP-resistant SKOV3 cells can effectively reverse the resistance of these cells to DDP and enhance the sensitivity to chemotherapy. |
| format | Article |
| id | doaj-art-9c3ae942344140a4a357139ce1c67857 |
| institution | Kabale University |
| issn | 2096-529X |
| language | zho |
| publishDate | 2025-08-01 |
| publisher | Editorial Office of Journal of Precision Medicine |
| record_format | Article |
| series | 精准医学杂志 |
| spelling | doaj-art-9c3ae942344140a4a357139ce1c678572025-08-20T04:00:28ZzhoEditorial Office of Journal of Precision Medicine精准医学杂志2096-529X2025-08-0140436236810.13362/j.jpmed.202540087The impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapyLIU Hui, CHEN Yunqing, QI Huiyang, SUN Xin, LOU Yanhui0Department of Gynecology, The Affiliated Hospital of Qingdao University, Qingdao 266003, ChinaObjective To investigate the impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy (NACT). Methods Tumor biopsy tissue samples were collected before NACT from 47 patients with advanced high-grade serous ovarian cancer (HGSOC) who underwent NACT and intermediate tumor cell reduction surgery in Department of Gynecology, The Affiliated Hospital of Qingdao University, from June 2017 to June 2019. Immunohistochemistry was used to measure the expression level of RAD51 protein in tumor tissue, and the correlation between its expression level and the sensitivity to NACT was analyzed. SKOV3 cells were divided into groups a-f, and cisplatin (DDP)-resistant SKOV3 cells were divided into groups A-F; the cells in groups b-f or groups B-F were transfected with NC-siRNA, siRNA-RAD51-264, siRNA-RAD51-416, siRNA-RAD51-608, and siRNA-RAD51-807, respectively, and then quantitative real-time PCR was used to measure the mRNA expression levels of RAD51 in each group of SKOV3 cells and DDP-resistant SKOV3 cells. The DDP-resistant SKOV3 cells were divided into SKOV3-Control group (control group), SKOV3-DDP-Control group (DDP control group), SKOV3-DDP-NC-siRNA group (transfected with NC-siRNA), and SKOV3-DDP-siRNA-RAD51 group (transfected with siRNA-RAD51), and Western blotting was used to measure the expression level of RAD51 protein in each group of cells. The DDP-resistant SKOV3 cells were divided into NC-siRNA group (transfected with NC-siRNA) and siRNA-RAD51 group (transfected with siRNA-RAD51-264), and DDP at various concentrations (0.39, 0.78, 1.56, 3.12, 6.25, 12.50, and 25.00 mg/L) was added for culture for 48 h; CCK-8 assay was used to measure the viability of DDP-resistant SKOV3 cells after transfection. The DDP-resistant SKOV3 cells were divided into Control group, NC-siRNA group (transfected with NC-siRNA), siRNA-RAD51 group (transfected with siRNA-RAD51), and siRNA-RAD51+DDP group (transfected with siRNA-RAD51 and subsequently treated with DDP), and flow cytometry was used to observe cell cycle alterations and apoptosis rate in DDP-resistant SKOV3 cells. Results Immunohistochemistry showed that there were no significant differences in age, FIGO stage (stage Ⅲ or Ⅳ), lymph node metastasis, ascites, and initial serum CA125 level between the patients with high RAD51 expression and those with low expression (P>0.05), and there was a significant difference in the expression level of RAD51 protein in tumor tissue between the chemotherapy-sensitive group and the insensitive group (χ2=10.85,P<0.05). RT-qPCR showed that the relative mRNA expression level of RAD51 in SKOV3-DDP cells was significantly higher than that in SKOV3 cells (t=6.73,P<0.05), and the mRNA expression level of RAD51 in group c/C was significantly lower than that in group a/A (F=7.81,6.58,P<0.05); there was no significant difference in the mRNA expression level of RAD51 between groups d/D, e/E, f/F and group a/A (P>0.05). Western blotting showed that compared with group A, group B had a significant increase in the protein expression level of RAD51 in cells, and compared with groups A and B, group D had a significant reduction in the protein expression level of RAD51 (F=10.56,P<0.05). CCK-8 assay showed that compared with the NC-siRNA group, the siRNA-RAD51-264 group had a significant reduction in cell viability at DDP concentrations of 3.12, 6.25, 12.50, and 25.00 mg/L (t=3.72-63.90,P<0.05). Flow cytometry showed that compared with the Control group and the NC-siRNA group, the siRNA-RAD51 group and the siRNA-RAD51+DDP group had a significant increase in the proportion of cells in the G0/G1 phase, and compared with the siRNA-RAD51 group, the siRNA-RAD51+DDP group had a significant increase in the proportion of cells in the G0/G1 phase (F=23.29,P<0.05); compared with the Control group and the NC-siRNA group, the siRNA-RAD51+DDP group had a significant reduction in the proportion of cells in the S phase (F=12.95,P<0.05). Compared with the Control group and the NC-siRNA group, both the siRNA-RAD51 group and the siRNA-RAD51+DDP group had a significant increase in apoptosis rate (F=16.34,P<0.05). Conclusion Patients with a high expression level of RAD51 protein in ovarian cancer tissue tend to have poorer sensitivity to NACT, and knocking down the expression of RAD51 gene in DDP-resistant SKOV3 cells can effectively reverse the resistance of these cells to DDP and enhance the sensitivity to chemotherapy.https://jpmed.qdu.edu.cn/fileup/2096-529X/PDF/1754471601716-1632270267.pdfcystadenocarcinoma, serous|ovarian neoplasms|chemotherapy, adjuvant|rad51 recombinase|drug resistance, neoplasm|cisplatin |
| spellingShingle | LIU Hui, CHEN Yunqing, QI Huiyang, SUN Xin, LOU Yanhui The impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy 精准医学杂志 cystadenocarcinoma, serous|ovarian neoplasms|chemotherapy, adjuvant|rad51 recombinase|drug resistance, neoplasm|cisplatin |
| title | The impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy |
| title_full | The impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy |
| title_fullStr | The impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy |
| title_full_unstemmed | The impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy |
| title_short | The impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy |
| title_sort | impact of rad51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy |
| topic | cystadenocarcinoma, serous|ovarian neoplasms|chemotherapy, adjuvant|rad51 recombinase|drug resistance, neoplasm|cisplatin |
| url | https://jpmed.qdu.edu.cn/fileup/2096-529X/PDF/1754471601716-1632270267.pdf |
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