Department of Gynecology and Obstetrics, The Second Affiliated Hospital of Soochow University, Suzhou, China.
Department of Gynecology, Shanghai Changning Maternity and Infant Health Hospital, Shanghai, China.
Cells Tissues Organs. 2022;211(1):16-29. doi: 10.1159/000519844. Epub 2021 Sep 27.
Long non-coding RNA HOTAIR has been revealed to participate in the tumorigenesis of various cancers. However, the mechanism of HOTAIR involvement in cervical cancer has not been identified. Hence, this study aimed to explore the oncogenic and chemoresistant roles of HOTAIR in cervical cancer, and its underlying mechanism. RT-PCR, Western blot, and Luciferase assay were employed to determine the relationship of HOTAIR with miR-29b and PTEN and to study the role of HOTAIR in cervical cancer. CCK8 assay, cell migration, and invasion assay were used to reveal the role of HOTAIR in cervical cancer cell proliferation and metastasis. The inhibitory dose of chemotherapeutics was determined by CCK8 assay using probit analysis. HOTAIR was found to bind with miR-29b, and a negative correlation existed between HOTAIR and miR-29b expression in cervical cancer cells. In addition, HOTAIR promoted the migration and proliferation of cervical cancer cell lines HeLa and Siha, showing effects opposite to miR-29b. Further, HOTAIR facilitated the resistance of both HeLa and Siha cells against cisplatin, paclitaxel and docetaxel, whereas miR-29 suppressed the resistance of both cervical cancer cells against the 3chemotherapeutics. In addition, HOTAIR enhanced epithelial-to-mesenchymal transition (EMT), while miR-29b exerted an inhibitory effect on EMT. In cervical cancer cells, miR-29b did not affect promoter methylation of PTEN but regulated PTEN expression by targeting SP1. Transfection of miR-29b mimics led to a significant downregulation of PI3K. HOTAIR promotes chemoresistance by facilitating EMT through the miR-29b/PTEN/PI3K axis in cervical cancer.
长链非编码 RNA HOTAIR 已被揭示参与多种癌症的发生。然而，HOTAIR 参与宫颈癌的机制尚未确定。因此，本研究旨在探讨 HOTAIR 在宫颈癌中的致癌和化疗耐药作用及其潜在机制。RT-PCR、Western blot 和 Luciferase 测定用于确定 HOTAIR 与 miR-29b 和 PTEN 的关系，并研究 HOTAIR 在宫颈癌中的作用。CCK8 测定、细胞迁移和侵袭测定用于揭示 HOTAIR 在宫颈癌细胞增殖和转移中的作用。使用概率分析的 CCK8 测定确定化疗药物的抑制剂量。发现 HOTAIR 与 miR-29b 结合，并且宫颈癌细胞中 HOTAIR 和 miR-29b 的表达呈负相关。此外，HOTAIR 促进了宫颈癌细胞系 HeLa 和 Siha 的迁移和增殖，表现出与 miR-29b 相反的作用。此外，HOTAIR 促进了 HeLa 和 Siha 细胞对顺铂、紫杉醇和多西紫杉醇的耐药性，而 miR-29b 抑制了这两种宫颈癌细胞对 3 种化疗药物的耐药性。此外，HOTAIR 增强了上皮间质转化 (EMT)，而 miR-29b 对 EMT 具有抑制作用。在宫颈癌细胞中，miR-29b 不影响 PTEN 启动子的甲基化，但通过靶向 SP1 调节 PTEN 的表达。miR-29b 模拟物的转染导致 PI3K 的显著下调。HOTAIR 通过 miR-29b/PTEN/PI3K 轴促进 EMT 从而促进宫颈癌的化疗耐药性。
