HOXC6在头颈部鳞状细胞癌中表达情况及其对体外细胞增殖、凋亡水平的影响

Expression of HOXC6 in the head and neck squamous cell carcinoma and its effect on cell proliferation and apoptosis levels in vitro

目的探究HOXC6在头颈部鳞状细胞癌中的表达情况及临床意义。方法通过TCGA数据库分析HOXC6基因在头颈鳞癌中的表达差异,并通过RT-qPCR检测临床头颈鳞癌标本中HOXC6的表达水平,通过Kaplan-Meier法分析HOXC6与头颈鳞癌患者临床预后的相关性,通过下调HOXC6在头颈鳞癌细胞株FaDu、TU-212中的表达,用CCK-8法及克隆形成实验检测头颈鳞癌细胞增殖和克隆形成能力,流式细胞术Annexin V-APC/7AAD双染色法检测癌细胞凋亡率,Western blot检测凋亡蛋白Caspase3、Caspase9的表达。结果生物信息学分析TCGA数据库得到HOXC6在头颈鳞癌中高表达(P<0.001),临床标本中HOXC6在头颈鳞癌组织中相对于癌旁组织高表达(P<0.001);预后分析发现HOXC6高表达患者总生存期明显下降,风险因子提高了0.42倍(P=0.01)且HOXC6高、低表达组在肿瘤的组织学分级上有差异(P<0.001)、在肿瘤淋巴结侵犯上有差异(P<0.05)、在总体生存期上有差异(P<0.01)。细胞实验证实下调头颈鳞癌细胞HOXC6表达可显著抑制癌细胞增殖和细胞克隆形成能力,同时癌细胞凋亡率上升且凋亡蛋白Caspase3及Caspase9剪接体表达增加。结论HOXC6在头颈鳞癌中高表达,并促进头颈鳞癌的增殖,抑制头颈鳞癌的凋亡,从而导致不良预后。

Objective To explore the expression,prognosis and clinical significance of HOXC6 in head and neck squamous cell carcinoma(HNSCC).Methods The TCGA database was used to analyze the expression difference of HOXC6 gene in head and neck squamous cell carcinoma,and the expression level of HOXC6 in clinical head and neck squamous cell carcinoma specimens was detected by RT-qPCR,and the correlation between HOXC6 and the clinical prognosis of head and neck squamous cell carcinoma patients was analyzed by Kaplan-Meier method,and the expression of HOXC6 in the head and neck squamous cell lines FaDu and TU-212 was down-regulated.The effect of HOXC6 on the proliferation and clonal formation of head and neck squamous cell carcinoma cells was detected by CCK-8 method and cloning formation experiment,the apoptosis rate of cancer cells was detected by flow cytometry Annexin V-APC/7AAD double staining,and the expression of apoptotic proteins Caspase3 and Caspase9 was detected by Western blot.Results Bioinformatics analysis of TCGA database showed that HOXC6 was highly expressed in head and neck squamous cell carcinoma(P<0.001),and HOXC6 was highly expressed in head and neck squamous cell carcinoma tissues relative to adjacent tissues(P<0.001).Prognostic analysis showed that the overall survival of patients with high HOXC6 expression was significantly decreased,and the risk factor was increased by 0.42 times(P=0.01).There were differences in histological grade of tumors(P<0.001),lymph node invasion(P<0.05),and overall survival(P<0.01)between the high and low HOXC6 expression groups.Cell experiments confirmed that downregulation of HOXC6 expression in head and neck squamous cell carcinoma cells could significantly inhibit cancer cell proliferation and cell clonal formation,while the apoptosis rate of cancer cells increased and the expression of apoptotic proteins Caspase3 and Caspase9 splicing bodies increased.Conclusion HOXC6 is highly expressed in HNSCC and promotes proliferation and suppresses apoptosis in HNSCC,thus leading to poor prognosis.