Department of Obstetrics and Gynecology, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, No. 32 Yihuan Road, Chengdu 610072, Sichuan, China.
Biol Chem. 2018 Apr 25;399(5):485-497. doi: 10.1515/hsz-2017-0274.
This study aimed at investigating the biological functions of long non-coding RNAs (lncRNAs) hox transcript antisense intergenic RNA (HOTAIR) in resistant ovarian cancer cells, exploring the regulation effect of HOTAIR on HOXA7, and investigating their influence on the chemosensitivity of ovarian cancer cells. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied for the verification of HOTAIR expression in resistant and sensitive groups. How HOTAIR downregulation affected cell proliferation, migration and invasion, and apoptosis were determined using the MTT assay and the colony formation assay, the Transwell assay and flow cytometry analysis, respectively. Immunohistochemistry was used to inspect the protein expression of HOXA7 in resistant and sensitive ovarian cancer tissues. The regulation relationship between HOTAIR and HOXA7 was investigated by qRT-PCR and Western blot. The effect of HOTAIR and HOXA7 on tumor growth was confirmed by the tumor xenograft model of nude mice. By knocking down HOXA7, HOTAIR downregulation restrained the ovarian cancer deterioration in functional experiments. Silencing of HOTAIR and HOXA7 could effectively inhibit tumor growth and increase chemosensitivity of ovarian tumors in nude mice. Downregulation of HOTAIR negatively affected the survival and activity of resistant ovarian cancer cells, and suppressed the expression of HOXA7. Silencing of HOTAIR and HOXA7 could increase the chemosensitivity of ovarian cancer cells, thus suppressing tumor development.
本研究旨在探讨长非编码 RNA（lncRNA）HOTAIR 在耐药卵巢癌细胞中的生物学功能，研究 HOTAIR 对 HOXA7 的调控作用及其对卵巢癌细胞化疗敏感性的影响。采用实时定量聚合酶链反应（qRT-PCR）验证耐药组和敏感组中 HOTAIR 的表达。通过 MTT 检测和集落形成实验、Transwell 检测和流式细胞术分析分别检测 HOTAIR 下调对细胞增殖、迁移和侵袭以及细胞凋亡的影响。免疫组织化学检测耐药和敏感卵巢癌组织中 HOXA7 的蛋白表达。通过 qRT-PCR 和 Western blot 研究 HOTAIR 和 HOXA7 之间的调控关系。通过裸鼠卵巢癌移植瘤模型证实 HOTAIR 和 HOXA7 对肿瘤生长的影响。敲低 HOXA7 可抑制功能实验中卵巢癌的恶化。沉默 HOTAIR 和 HOXA7 可有效抑制裸鼠卵巢肿瘤的生长并增加其化疗敏感性。下调 HOTAIR 可负性影响耐药卵巢癌细胞的存活和活性，并抑制 HOXA7 的表达。沉默 HOTAIR 和 HOXA7 可增加卵巢癌细胞的化疗敏感性，从而抑制肿瘤的发展。
