Rui Yan, Peng Wen-Jia, Wang Ming, Wang Qian, Liu Zi-Li, Chen Yu-Qing, Huang Li-Nian
Department of Respiration and Critical Care Medicine, The First Affiliated Hospital of Bengbu Medical College, Lung Cancer Diagnosis and Treatment Center of Anhui Province, Anhui Provincial Key Laboratory of Clinical Basic Research on Respiratory Disease, Bengbu, Anhui 233004, P.R. China.
Department of Epidemiology and Health Statistics, Bengbu Medical College, Bengbu, Anhui 233000, P.R. China.
Int J Oncol. 2017 Mar;50(3):815-822. doi: 10.3892/ijo.2017.3856. Epub 2017 Jan 19.
HIST1H3D gene encodes histone H3.1 and is involved in gene-silencing and heterochromatin formation. HIST1H3D expression is upregulated in primary gastric cancer tissue. In this study, we explored the effects of HIST1H3D expression on lung cancer, and its mechanisms. HIST1H3D expression was measured by immunohistochemistry and RT-PCR in lung cancer tissues and human lung cancer cell lines. Cell proliferation was assessed by MTT assay. Flow cytometric analysis was used to determine cell cycle distribution and apoptosis. Levels of related proteins were detected by western blotting. Bioinformatics analysis was performed to investigate related signaling pathways. cDNA microarray analysis was performed to identify differentially expressed genes following HIST1H3D knockdown. HIST1H3D expression was upregulated in lung cancer tissue samples and the H1299 human lung cancer cell line (P<0.01). Regulation of HIST1H3D expression in nucleus of cells in lung cancer tissues was significant associated with tumor stage (P=0.02) and lymph node metastases (P=0.04). Downregulation of HIST1H3D expression led to suppression of proliferation and colony forming ability, cell cycle arrest at the G0/G1 phase, and promotion of cell apoptosis. The microarray data revealed 522 genes that were differentially expressed after HIST1H3D knockdown in H1299 cells. These genes were shown to be linked to numerous pathways, including the cell cycle, p53 signaling, and MCM. Western blot analysis confirmed upregulated expression of the THBS1 and TP53I3 genes, and downregulated expression of the CDK6, CDKN1 and CCNE2 genes. In conclusion, our results suggest that HIST1H3D is highly expressed in lung cancer cell lines and tissues. Furthermore, HIST1H3D may be important in cell proliferation, apoptosis and cell cycle progression, and is implicated as a potential therapeutic target for lung cancer.
HIST1H3D基因编码组蛋白H3.1，并参与基因沉默和异染色质形成。HIST1H3D在原发性胃癌组织中表达上调。在本研究中，我们探讨了HIST1H3D表达对肺癌的影响及其机制。通过免疫组织化学和RT-PCR检测肺癌组织和人肺癌细胞系中的HIST1H3D表达。通过MTT法评估细胞增殖。采用流式细胞术分析确定细胞周期分布和凋亡情况。通过蛋白质印迹法检测相关蛋白水平。进行生物信息学分析以研究相关信号通路。进行cDNA微阵列分析以鉴定HIST1H3D敲低后差异表达的基因。HIST1H3D在肺癌组织样本和H1299人肺癌细胞系中表达上调（P<0.01）。肺癌组织细胞核中HIST1H3D表达的调节与肿瘤分期（P=0.02）和淋巴结转移（P=0.04）显著相关。HIST1H3D表达下调导致增殖和集落形成能力受到抑制，细胞周期停滞在G0/G1期，并促进细胞凋亡。微阵列数据显示，H1299细胞中HIST1H3D敲低后有522个基因差异表达。这些基因被证明与众多通路相关，包括细胞周期、p53信号通路和MCM。蛋白质印迹分析证实THBS1和TP53I3基因表达上调，CDK6、CDKN1和CCNE2基因表达下调。总之，我们的结果表明HIST1H3D在肺癌细胞系和组织中高表达。此外，HIST1H3D可能在细胞增殖、凋亡和细胞周期进程中起重要作用，并被认为是肺癌的潜在治疗靶点。
