| 王 充,杨诗敏,张春晓,周冬梅,向江东,席晓薇.m6A 去甲基化酶ALKBH5 对上皮性卵巢癌恶性生物学行为的影响及其作用机制研究[J].现代生物医学进展英文版,2023,(17):3201-3210. |
| m6A 去甲基化酶ALKBH5 对上皮性卵巢癌恶性生物学行为的影响及其作用机制研究 |
| Effects of m6A Demethylase ALKBH5 on the Malignant Biological Behavior and Mechanism Action of Epithelial Ovarian Cancer |
| Received:March 06, 2023 Revised:March 28, 2023 |
| DOI：10.13241/j.cnki.pmb.2023.17.001 |
| 中文关键词: m6A ALKBH5 上皮性卵巢癌 |
| 英文关键词: m6A ALKBH5 Epithelial ovarian cancer |
| 基金项目:国家自然科学基金项目（81772767） |
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| 中文摘要: |
| 摘要 目的：ALKBH5最近被证明是与各种癌症相关的RNA N6-甲基腺苷（m6A）去甲基转移酶之一，但其在上皮性卵巢癌中的作用缺乏研究。本研究旨在探讨ALKBH5在上皮性卵巢癌中的机制。方法：利用蛋白免疫印迹、免疫组化检测ALKBH5在卵巢癌细胞和组织中的表达情况，并分析其与临床病理特征和预后关系；通过慢病毒感染构建ALKBH5稳定过表达或敲减卵巢癌细胞系，采用CCK-8、平板克隆、划痕、Transwell 迁移和侵袭等实验研究ALKBH5 对细胞增殖、迁移及侵袭能力的影响；通过蛋白免疫印迹和斑点印迹实验，探究ALKBH5-HIF-1α环的相互作用情况。结果：ALKBH5在卵巢癌细胞系和卵巢癌组织中高表达，ALKBH5蛋白在卵巢癌组织中的表达与CA125水平、FIGO分期和组织学类型相关，且高表达的ALKBH5水平与较差的预后相关。通过慢病毒感染构建出可靠的ALKBH5敲减/过表达稳转细胞系，CCK-8、平板克隆实验表明敲减ALKBH5降低卵巢癌细胞体外增殖速率和集落形成，划痕、Transwell 迁移和侵袭实验表明敲减ALKBH5可抑制卵巢癌细胞体外迁移和侵袭能力。与对照组相比，过表达ALKBH5增强卵巢癌细胞体外增殖速率、集落形成、迁移和侵袭能力。进一步的实验发现ALKBH5和HIF-1α互相促进表达。结论：m6A去甲基化酶ALKBH5在上皮性卵巢癌中高表达且与预后相关，通过促进卵巢癌细胞增殖、迁移及侵袭发挥致癌基因的作用。因此，抑制ALKBH5的表达可能是靶向治疗上皮性卵巢癌的潜在策略。 |
| 英文摘要: |
| ABSTRACT Objective: ALKBH5 has recently been identified as one of the RNA N6-methyladenosine (m6A) demethylases associated with various types of cancer. However, its role in epithelial ovarian cancer remains unclear. This study aimed to investigate the mechanism of ALKBH5 in epithelial ovarian cancer. Methods: The expression of ALKBH5 in ovarian cancer cells and tissues was detected by western blotting and immunohistochemistry, respectively. The online Kaplan-Meier survival analysis was used to explore the correlation between ALKBH5 and ovarian cancer prognosis and to analyze its relationship with clinicopathological features and prognosis. Stable ovarian cancer cell lines with overexpression or knockdown of ALKBH5 were constructed by lentivirus infection. The function of ALKBH5 was investigated in vitro by using CCK-8, colony formation assay, wound healing assay, Transwell migration and invasion assay and other experimental methods to study the effects of ALKBH5 on cell proliferation, migration and invasion. Ovarian cancer cells were treated with hypoxia and the interaction between ALKBH5-HIF-1α loop was explored by western blotting and dot blotting experiments. Results: ALKBH5 was highly expressed in ovarian cancer cell lines and ovarian cancer tissues. The expression of ALKBH5 protein in ovarian cancer tissues was correlated with CA125 level, FIGO stage and histological type, and high expression of ALKBH5 level was associated with poor prognosis. Reliable ALKBH5 knockdown/overexpression stable cell lines were constructed by lentivirus infection. CCK-8 and colony formation assay showed that knockdown of ALKBH5 reduced the proliferation rate and colony formation of ovarian cancer cells in vitro. Wound healing and Transwell migration and invasion assay showed that knockdown of ALKBH5 inhibited the migration and invasion abilities of ovarian cancer cells in vitro. In vitro experiments confirmed that ALKBH5 promoted proliferation, migration and invasion in ovarian cancer. Compared with the control group, overexpression of ALKBH5 enhanced the proliferation rate, colony formation, migration and invasion abilities of ovarian cancer cells in vitro. Further experiments revealed that ALKBH5 and HIF-1α positively regulated each other's expression. Conclusion: m6A demethylase ALKBH5 is highly expressed and associated with prognosis in epithelial ovarian cancer, and plays an oncogenic role by promoting ovarian cancer cell proliferation, migration and invasion. Therefore, inhibiting the expression of ALKBH5 may be a potential strategy for targeted therapy of epithelial ovarian cancer. |
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