Department of Urology, Xiangya Hospital, Central South University, Changsha, China.
J Cell Biochem. 2019 Jun;120(6):8965-8974. doi: 10.1002/jcb.27851. Epub 2019 Feb 27.
Prostate cancer (PCa) is a prevalent and deadly cancer worldwide. Considering the malignant progression and therapeutic resistance of PCa, further dissection of the underlying mechanisms and exploration of novel therapeutic targets for PCa are urgently needed. The long noncoding RNA HOTTIP has recently been revealed as an oncogenic regulator in different cancers; however, whether HOTTIP is involved in PCa remains poorly understood. Here, we examined the crucial roles of HOTTIP in the proliferation and chemoresistance of PCa.
Quantitative real-time PCR (qRT-PCR) was performed to detect the HOTTIP messenger RNA (mRNA) levels in PCa samples from patients and PCa cells. Then, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation, and cell cycle and flow cytometry assays were performed to investigate the proliferation and cisplatin-resistance of PCa cells with silenced HOTTIP compared with a negative control. We applied Western blotting, qRT-PCR and a TOP/FOP assay to explore the relevant mechanisms.
In this study, we found that the HOTTIP mRNA levels were increased in the PCa patient samples and PCa cell lines compared with the controls. The knockdown of HOTTIP not only inhibited the proliferation of PCa cells but also facilitated cell cycle arrest and chemosensitivity to cisplatin. Furthermore, the qRT-PCR, Western blotting, TOP/FOP assays, MTT assay, and flow cytometry revealed that Wnt/β-catenin signaling was related to the regulation of HOTTIP in cell proliferation, cell cycle arrest, and chemoresistance to cisplatin in PCa.
Taken together, our findings suggest that HOTTIP may be a potent therapeutic target for PCa, and HOTTIP inhibitors might be regarded as effective strategies for PCa therapy.
前列腺癌（PCa）是一种在全球范围内普遍存在且致命的癌症。鉴于 PCa 的恶性进展和治疗抵抗，迫切需要进一步剖析其潜在机制，并探索用于 PCa 的新型治疗靶点。长链非编码 RNA HOTTIP 最近被揭示为不同癌症中的致癌调节剂；然而，HOTTIP 是否参与 PCa 仍知之甚少。在此，我们研究了 HOTTIP 在 PCa 增殖和化疗耐药中的关键作用。
采用实时定量 PCR（qRT-PCR）检测来自患者的 PCa 样本和 PCa 细胞中的 HOTTIP 信使 RNA（mRNA）水平。然后，通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）、集落形成、细胞周期和流式细胞术检测与阴性对照相比沉默 HOTTIP 的 PCa 细胞的增殖和顺铂耐药性。我们应用 Western blot、qRT-PCR 和 TOP/FOP 测定来探索相关机制。
在本研究中，我们发现与对照相比，HOTTIP mRNA 水平在 PCa 患者样本和 PCa 细胞系中升高。HOTTIP 的敲低不仅抑制了 PCa 细胞的增殖，而且促进了细胞周期停滞和对顺铂的化疗敏感性。此外，qRT-PCR、Western blot、TOP/FOP 测定、MTT 测定和流式细胞术表明，Wnt/β-catenin 信号通路与 HOTTIP 调节 PCa 细胞增殖、细胞周期停滞和对顺铂的化疗耐药性有关。
综上所述，我们的研究结果表明 HOTTIP 可能是 PCa 的一个有潜力的治疗靶点，HOTTIP 抑制剂可能被视为 PCa 治疗的有效策略。
