Ming Fei, Sun Qianqiang
Department of Thoracic Surgery, Hubei Cancer Hospital, Wuhan, Hubei 430000, P.R. China.
Mol Med Rep. 2017 Jul;16(1):746-754. doi: 10.3892/mmr.2017.6665. Epub 2017 May 31.
Protein tyrosine phosphatase receptor‑type O (PTPRO), a member of the PTP family, has been frequently reported as potential tumor suppressor in many types of cancer. However, the exact function of PTPRO in lung squamous cell carcinoma (LSCC) remains unclear. Bisulfite sequencing and methylation specific polymerase chain reaction (PCR) were used to identify the methylation status of PTPRO in LSCC cells, and quantitative methylation specific PCR was used to evaluate the methylation levels of PTPRO in LSCC patients. Stably expressing PTPRO vectors were constructed and transfected into H520 and SK‑MES‑1 cells, followed by MTT and colony formation assays, and analysis of tumor weight and volume in in vivo mouse xenograft models. The present study demonstrated that the CpG island of PTPRO exon 1 was obviously hypermethylated in LSCC cells and tissues. The mRNA expression of PTPRO could be restored by treatment with a demethylation agent. Increased methylation and decreased mRNA levels of PTPRO were observed in LSCC samples compared with adjacent healthy tissues, and were associated with poor prognosis of patients. The mRNA expression of PTPRO was negatively correlated with its methylation level in tumors. Functionally, ectopic PTPRO expression in LSCC cells significantly inhibited the proliferation rates, and colony formation, in comparison with control and non‑transfected cells. In vivo assays confirmed the inhibitory effect of PTPRO on LSCC cell growth. In conclusion, these data provided evidence that epigenetic regulation of PTPRO impairs its tumor suppressor role in LSCC, and restoration of PTPRO may be a potential therapeutic strategy.
蛋白酪氨酸磷酸酶受体O型(PTPRO)是蛋白酪氨酸磷酸酶(PTP)家族的成员之一,在多种癌症中常被报道为潜在的肿瘤抑制因子。然而,PTPRO在肺鳞状细胞癌(LSCC)中的具体功能仍不清楚。采用亚硫酸氢盐测序和甲基化特异性聚合酶链反应(PCR)来鉴定LSCC细胞中PTPRO的甲基化状态,并使用定量甲基化特异性PCR评估LSCC患者中PTPRO的甲基化水平。构建稳定表达PTPRO的载体并将其转染到H520和SK-MES-1细胞中,随后进行MTT和集落形成试验,并在体内小鼠异种移植模型中分析肿瘤重量和体积。本研究表明,LSCC细胞和组织中PTPRO外显子1的CpG岛明显高度甲基化。用去甲基化剂处理可恢复PTPRO的mRNA表达。与相邻的健康组织相比,在LSCC样本中观察到PTPRO的甲基化增加和mRNA水平降低,并且这与患者的不良预后相关。肿瘤中PTPRO的mRNA表达与其甲基化水平呈负相关。在功能上,与对照细胞和未转染细胞相比,LSCC细胞中异位表达PTPRO可显著抑制增殖率和集落形成。体内试验证实了PTPRO对LSCC细胞生长的抑制作用。总之,这些数据表明PTPRO的表观遗传调控损害了其在LSCC中的肿瘤抑制作用,恢复PTPRO可能是一种潜在的治疗策略。