PAK2在喉鳞状细胞癌中的表达及其与患者临床病理特征、化疗敏感性的关系

Expression of p21-activated kinase 2 in laryngeal squamous cell carcinoma and its correlation with clinicopathological characteristics and chemosensitivity

目的探讨p21活化激酶2(PAK2)在喉鳞状细胞癌中的表达情况及其与患者临床病理特征、化疗敏感性的关系。方法从癌症基因组图谱(TCGA)数据库中下载喉鳞状细胞癌相关转录组测序(RNA-seq)数据,123例患者列入研究(12例有癌组织和正常组织数据,其余111例仅有癌组织数据);应用R软件分析PAK2在癌和癌旁组织中的差异表达,应用京都基因与基因组百科全书(KEGG)数据库信号通路富集分析PAK2在喉鳞状细胞癌中的潜在功能。回顾性选取2016年4月至2021年6月于朝阳市中心医院行手术切除的原发性喉鳞状细胞癌组织、对应癌旁组织标本各34例,另取喉正常黏膜组织20例作为对照;应用免疫组织化学法检测各组织中PAK2表达情况,分析其与临床病理因素的相关性。回顾性选取同期诱导化疗前行组织标本采集的声门上型原发性喉鳞状细胞癌患者35例,包括20例化疗敏感患者和15例化疗耐药患者;应用实时荧光定量聚合酶链反应(qRT-PCR)检测癌组织中PAK2 mRNA相对表达量。结果TCGA数据库数据分析显示,与癌旁组织相比,癌组织中PAK2表达升高(P=0.012);KEGG数据库信号通路分析显示,喉鳞状细胞癌PAK2高表达与信号转导通路、细胞周期、癌症等相关。免疫组织化学法检测显示,34例喉鳞状细胞癌组织中PAK2阳性患者比例高于癌旁组织和正常组织[58.82%(20/34)比0.03%(1/34)、0(0/20),均P<0.001];不同分化程度[高分化比低、中分化:33.33%(6/18)比87.50%(14/16)]、淋巴结转移情况[有比无:90.91%(10/11)比43.48%(10/23)]、TNM分期[Ⅲ~Ⅳ期比Ⅰ~Ⅱ期:82.35%(14/17)比35.29%(6/17)]患者间PAK2阳性患者比例差异均有统计学意义(均P<0.05),PAK2阳性与临床分型、肿瘤大小、吸烟史、饮酒史、年龄均无关(均P>0.05)。qRT-PCR检测显示,化疗耐药组癌组织PAK2 mRNA相对表达量高于化疗敏感组(3.89±0.12比0.78±0.23,P<0.001)。结论喉鳞状细胞癌组织PAK2表达水平升高,PAK2高表达与喉鳞状细胞癌患者恶性临床病理特征密切相关,其高表达可能提示对传统化疗方案不敏感;PAK2可能是靶向调节喉鳞状细胞癌化疗敏感性的潜在基因。

Objective To investigate the expression of p21-activated kinase 2(PAK2)in laryngeal squamous cell carcinoma and its relationship with the clinicopathological characteristics and chemosensitivity of patients.Methods Transcriptome sequencing(RNA-seq)data for laryngeal squamous cell carcinoma were downloaded from the Cancer Genome Atlas(TCGA)database,and 123 patients were included in the study(12 cases had cancer tissues and normal tissues data,and the remaining 111 only had cancer tissues data).Differential expression of PAK2 in cancer and para-cancer tissues was analyzed by using R software,and the potential function of PAK2 in laryngeal squamous cell carcinoma was investigated by using the Kyoto Encyclopedia of Genes and Genomes(KEGG)database signaling pathway enrichment.A total of 34 patients with primary laryngeal squamous cell carcinoma tissues and corresponding para-carcinoma 34 tissue specimens who underwent surgical resection were retrospectively selected from Chaoyang Central Hospital between April 2016 and June 2021,and 20 cases of normal laryngeal mucosa tissues were selected as the controls.Immunohistochemistry was used to detect the expression of PAK2 in various tissues,and its correlation with clinicopathological factors was analyzed.A total of 35 supraglottic primary laryngeal squamous cell carcinoma patients were retrospectively collected before induction chemotherapy during the same period,including 20 patients sensitive to chemotherapy and 15 patients resistant to chemotherapy.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the relative expression level of PAK2 mRNA in cancer tissues.Results Analysis of TCGA database data showed that PAK2 expression was increased in cancer tissues compared with that in para-cancer tissues(P=0.012);KEGG database signaling pathways showed that the high expression of PAK2 in laryngeal squamous cell carcinoma was related to signal transduction pathways,cell cycle,and cancer.Immunohistochemistry showed that the proportion of PAK2 positive in 34 cases of laryngeal squamous cell carcinoma tissues was higher than that in adjacent tissues and normal tissues[58.82%(20/34)vs.0.03%(1/34),0(0/20),all P<0.001].There were statistically significant differences in the proportion of PAK2 positive patients stratified with different degrees of differentiation[high differentiation vs.low or middle differentiation:33.33%(6/18)vs.87.50%(14/16)],lymph node metastasis[presence vs.absence:90.91%(10/11)vs.43.48%(10/23)],TNM staging[stageⅢ-Ⅳvs.stageⅠ-Ⅱ:82.35%(14/17)vs.35.29%(6/17)](all P<0.05),and PAK2 positive patients were not associated with clinical type,tumor size,smoking history,drinking history,and age(all P>0.05).qRT-PCR showed that the relative expression level of PAK2 mRNA in the chemotherapy-resistant group was higher than that in the chemotherapy-sensitive group(3.89±0.12 vs.0.78±0.23,P<0.001).Conclusions The expression level of PAK2 in laryngeal squamous cell carcinoma tissues is increased,and the high expression of PAK2 is closely related to the malignant clinical characteristics of patients with laryngeal squamous cell carcinoma.The high expression of PAK2 may indicate the insensitivity to traditional chemotherapy regimens,and PAK2 may be a potential gene that targets and regulates the chemosensitivity of laryngeal squamous cell carcinoma.