Roudkenar Mehryar Habibi, Fukumoto Motoi, Roushandeh Amaneh Mohammadi, Kuwahra Youshikazu, Uroshihara Yusuke, Harada Hiroshi, Fukumoto Manabu
Cardiovascular Diseases Research Center Department of Cardiology, Heshmat Hospital, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
Department of Pathology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Miyagi, 980-8575, Japan.
Cytotechnology. 2020 Feb;72(1):141-153. doi: 10.1007/s10616-019-00364-9. Epub 2020 Jan 8.
Cellular radioresistance is one of the major obstacles to the effectiveness of cancer radiotherapy. In an attempt to elucidate the implication of HIF-1α and miR-17-92 expressions in refractory radioresistant cells and also in order to study the potential applications of these molecules as novel therapeutic modalities to overcome radioresistant cancers, the current study was conducted. Clinically relevant radioresistant (CRR) cells from human cancer cell lines were established by exposing to long-term fractionated radiation of X-rays. Correspondingly, microarray analysis and real time RT-PCR were performed to find miRNA involved in the CRR phenotype. HIF-1α was down-regulated and miR17-92 cluster was overexpressed in CRR cells by transfection. The expression of miR 17-3p was inhibited by specific inhibitors and miR 19a was enforced by mimics, respectively in parental cells. Overexpression of HIF-1α in parental cells or down regulation of HIF-1α in CRR cells were not involved in radioresistance. However, when HIF-1α was genetically modified to constitutively express under normoxia condition, it was rendered for protection to cells. Exogenous overexpression of miR 17-92 cluster in CRR cells resulted in abolition of HIF-1α expression and restored sensitizations to ionizing radiation. Attenuated expression of miR-17-3p in parental cells protected them from irradiation. Overall, fine-tune deregulation of miR 17-92 cluster in CRR cells might account for the accumulation of HIF-1α in the CRR cells following exposure to irradiation.
细胞放射抗性是癌症放疗有效性的主要障碍之一。为了阐明缺氧诱导因子-1α(HIF-1α)和miR-17-92表达在难治性放射抗性细胞中的意义,并研究这些分子作为克服放射抗性癌症的新型治疗方式的潜在应用,开展了本研究。通过对人癌细胞系进行长期分次X射线辐射,建立了临床相关的放射抗性(CRR)细胞。相应地,进行了微阵列分析和实时逆转录聚合酶链反应(RT-PCR)以寻找参与CRR表型的微小RNA(miRNA)。通过转染,CRR细胞中HIF-1α表达下调,miR17-92簇过表达。分别在亲代细胞中用特异性抑制剂抑制miR 17-3p的表达,并用模拟物增强miR 19a的表达。亲代细胞中HIF-1α的过表达或CRR细胞中HIF-1α的下调与放射抗性无关。然而,当HIF-1α经基因改造在常氧条件下组成性表达时,它能保护细胞。CRR细胞中外源性过表达miR 17-92簇导致HIF-1α表达消失,并恢复对电离辐射的敏感性。亲代细胞中miR-17-3p表达减弱使其免受辐射。总体而言,CRR细胞中miR 17-92簇的微调失调可能是照射后CRR细胞中HIF-1α积累的原因。