| 王君霞,刘欣悦,刘杨钰,等.拓扑异构酶Ⅱα激活PI3K/AKT轴促进上皮间质转化并影响卵巢癌顺铂耐药性的研究[J].肿瘤学杂志,2026,32(5):375-384. |
| 拓扑异构酶Ⅱα激活PI3K/AKT轴促进上皮间质转化并影响卵巢癌顺铂耐药性的研究 |
| Topoisomerase Ⅱα Promotes Epithelial-Mesenchymal Transition and Enhances Cisplatin Resistance via PI3K/AKT Axis Activation in Ovarian Cancer |
| 投稿时间：2025-03-07 |
| DOI：10.11735/j.issn.1671-170X.2026.05.B004 |
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| 中文关键词: 卵巢肿瘤 上皮性肿瘤 拓扑异构酶 PI3K/AKT通路 上皮间质转化 顺铂 耐药 |
| 英文关键词:ovarian neoplasms epithelial neoplasms topoisomerase PI3K/AKT pathway epithelial-mesenchymal transition cisplatin drug resistance |
| 基金项目:国家自然科学基金（61975105）；山西省基础研究计划项目（202303021212366） |
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| 中文摘要: |
| 摘 要：［目的］ 研究拓扑异构酶Ⅱα（topoisomerase Ⅱ alpha，TOP2A）在上皮性卵巢癌组织中的表达及其对耐药上皮性卵巢癌细胞的生物学行为和顺铂敏感性的影响，并探究其作用机制。［方法］ 研究纳入2021—2024年山西医科大学附属山西省人民医院75例上皮性卵巢癌患者的卵巢癌组织和15例因非卵巢疾病（子宫腺肌病、子宫多发肌瘤）切除双侧附件的正常卵巢组织。采用免疫组化检测TOP2A在上皮性卵巢癌组织中的表达。蛋白印迹分析（Western blot）筛选TOP2A高表达的卵巢癌细胞株（OVCAR3、SKOV3）及其相应顺铂耐药细胞株（SKOV3/DDP、OVCAR3/DDP）作为研究对象，随后分别慢病毒构建TOP2A低表达的耐药细胞稳转株用于后续研究。实验分为空转对照组、TOP2A低表达组1及TOP2A低表达组2三组，上述两种细胞系均按此分组通过细胞计数试剂盒8实验、集落形成实验、细胞划痕实验、细胞侵袭实验、实时定量PCR以及Western blot实验检测细胞活力、增殖能力、迁移及侵袭能力、TOP2AmRNA及TOP2A蛋白水平表达和磷脂酰肌醇3激酶/蛋白激酶B（phosphoinositide 3 kinase/protein kinase B，PI3K/AKT）轴、上皮间质转化（epithelial-mesenchymal transition，EMT）的相关蛋白表达情况。随后使用PI3K激动剂（740Y-P）处理低表达TOP2A的耐药细胞稳转株（SKOV3/DDP、OVCAR3/DDP），Western blot检测PI3K、磷酸化PI3K及EMT相关标志蛋白的表达，进一步验证TOP2A通过PI3K/AKT通路影响耐药上皮性卵巢癌细胞恶性生物行为及耐药性。［结果］ 上皮性卵巢癌组织和卵巢癌耐药细胞中TOP2A表达水平升高，其表达程度与FIGO分期、分级及淋巴结转移呈正相关（P<0.05），与患者的年龄未见明显相关性（P>0.05）。在两种卵巢癌耐药细胞系中，TOP2A细胞活力、增殖能力、侵袭和迁移能力均下降（P<0.01），铂敏感性均上升（P<0.01），磷酸化PI3K、磷酸化AKT蛋白表达均下降（P<0.01）、上皮标志物E-cadherin蛋白表达均上升、间质标志物N-cadherin、Snail和Vimentin蛋白表达均下降（P<0.01）。加入PI3K激动剂740Y-P后，能够部分恢复磷酸化PI3K、磷酸化AKT和间质标志物N-cadherin、Snail和Vimentin水平，同时降低 E-钙黏蛋白的表达（P均<0.01）。［结论］ TOP2A在原发上皮性卵巢癌组织中异常高表达，并通过调控PI3K/AKT轴调节EMT过程，从而影响肿瘤细胞恶性生物学行为及EMT过程，进而影响卵巢癌顺铂敏感性。 |
| 英文摘要: |
| Abstract： ［Objective］ To investigate the expression of topoisomerase Ⅱ alpha (TOP2A) in epithelial ovarian cancer (EOC) tissues and its impact on the biological behavior and cisplatin sensitivity of drug-resistant EOC cells， as well as to explore the underlying mechanism. ［Methods］ A total of 75 ovarian cancer tissues from patients with EOC and 15 normal ovarian tissues from patients who underwent bilateral adnexectomy due to non-ovarian diseases (adenomyosis， multiple uterine fibroids) were included in the study from Shanxi Provincial People’s Hospital Affiliated to Shanxi Medical University from 2021 to 2024. Western blot was employed to screen EOC cell lines with high TOP2A expression (OVCAR3， SKOV3) and their corresponding cisplatin-resistant counterparts(SKOV3/DDP， OVCAR3/DDP) for further study. Lentiviral-mediated knockdown was then used to generate stable TOP2A-low-expressing cisplatin-resistant cell lines. The cells were divided into three groups： empty vector control， and two TOP2A knockdown groups. Cell viability， proliferation， migration， and invasion were assessed using cell counting kit 8， colony formation， wound healing，and Transwell assays， respectively. The expression levels of TOP2A， phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) pathway proteins， and epithelial-mesenchymal transition (EMT) markers were detected by real-time polymerase chain reaction and Western blot. Subsequently， TOP2A-knockdown cells were treated with the PI3K agonist 740Y-P， and the expression of related proteins was analyzed to further confirm that TOP2A affects malignant behaviors and drug resistance through the PI3K/AKT axis. ［Results］ TOP2A was significantly upregulated in EOC tissues and cisplatin-resistant cells. Its expression was correlated positively with FIGO stage， grade， and lymph node metastasis (P<0.05)， but not with patient age (P>0.05). Compared with the control group， TOP2A knockdown inhibited cell viability， proliferation， migration， and invasion， increased cisplatin sensitivity， downregulated phosphorylated PI3K (p-PI3K) and phosphorylated AKT (p-AKT)， upregulated the epithelial marker E-cadherin， and downregulated the mesenchymal markers N-cadherin， Snail， and Vimentin (all P<0.01). Treatment with 740Y-P in TOP2A-knockdown cells partially restored the levels of p-PI3K， p-AKT， and mesenchymal markers N-cadherin， Snail， and Vimentin， while reducing E-cadherin expression (all P<0.01). ［Conclusion］ TOP2A is aberrantly overexpressed in primary EOC tissues. It regulates the EMT process by modulating the PI3K/AKT axis， thereby influencing the malignant behaviors of tumor cells and ultimately contributing to cisplatin resistance in ovarian cancer. |
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