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慢病毒介导的MPHOSPH8沉默抑制甲状腺髓样癌增殖并增强凋亡。

Lentivirus-mediated silencing of MPHOSPH8 inhibits MTC proliferation and enhances apoptosis.

作者信息

Li Peiyong, Yang Weiping, Shen Baiyong, Li Hongwei, Yan Jiqi

机构信息

Department of Nuclear Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, P.R. China.

Department of Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, P.R. China.

出版信息

Oncol Lett. 2016 Jun;11(6):4117-4122. doi: 10.3892/ol.2016.4545. Epub 2016 May 6.

Abstract

Thyroid carcinoma (TC) is the most common malignancy of the endocrine organs, and its incidence rate has steadily increased over the last decade. For medullary thyroid cancer (MTC), a type of TC, a high mortality rate has been reported. In previous studies, M-phase phosphoprotein 8 (MPHOSPH8) displayed an elevated expression in various human carcinoma cells. Thus, MPHOSPH8 may be a sensitive biomarker that could be used for the diagnosis and follow-up of MTC. In the present study, plasmids of RNA interference targeting the MPHOSPH8 gene were constructed. Once these lentiviruses targeting MPHOSPH8 were transfected into the MTC cell line TT, cell viability and proliferation were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Flow cytometry was used to assess the cell cycle distribution and apoptosis. The expression levels of MPHOSPH8 were detected by reverse transcription quantitative-polymerase chain reaction and western blot analyses. Depletion of MPHOSPH8 significantly inhibited cell proliferation. Furthermore, knockdown of MPHOSPH8 in TT cells led to G0/G1 phase cell cycle arrest and apoptosis. The results of the present study suggest that MPHOSPH8 promotes cell proliferation and may be a potential target for anticancer therapy of MTC.

摘要

甲状腺癌(TC)是内分泌器官最常见的恶性肿瘤,其发病率在过去十年中稳步上升。对于甲状腺髓样癌(MTC)这种TC类型,已有报道称其死亡率很高。在先前的研究中,M期磷蛋白8(MPHOSPH8)在各种人类癌细胞中表达升高。因此,MPHOSPH8可能是一种可用于MTC诊断和随访的敏感生物标志物。在本研究中,构建了靶向MPHOSPH8基因的RNA干扰质粒。一旦将这些靶向MPHOSPH8的慢病毒转染到MTC细胞系TT中,就通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法测量细胞活力和增殖。使用流式细胞术评估细胞周期分布和凋亡。通过逆转录定量聚合酶链反应和蛋白质免疫印迹分析检测MPHOSPH8的表达水平。MPHOSPH8的缺失显著抑制细胞增殖。此外,在TT细胞中敲低MPHOSPH8导致G0/G1期细胞周期停滞和凋亡。本研究结果表明,MPHOSPH8促进细胞增殖,可能是MTC抗癌治疗的潜在靶点。

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