Objective: To analyze the differentially expressed genes related to the chemosensitivity with the TPF regimen for hypopharyngeal squamous cell carcinoma and to measure potential functional targeting genes expressions. Methods: Twenty-nine patients with primary hypopharyngeal cancer who underwent induction chemotherapy with TPF from January 2013 to December 2017 in Beijing Tongren Hospital were enrolled for microarray analysis, including 28 males and 1 female, aged from 43 to 73 years old. Among them, 16 patients were sensitive to chemotherapy while 13 patients were non-sensitive. Illumina Human HT-12 Bead Chip was applied to analyze the gene expressions and online bioinformatics analysis was used to analyze the differentially expressed genes. Reverse transcription and quantitative real-time PCR (RT-qPCR) was used to measure the mRNA expression of potential functional genes of TPF induction chemotherapy in 43 samples, 29 from original patients and 14 from additional patients. Graphpad prism 7.0 software was used for statistical analysis. Results: A total of 1 381 significantly differentially expressed genes were screened out. By GO analysis, up-regulated genes included sequestering in extracellular matrix, chemokine receptor binding and potassium channel regulator activity; down-regulated genes included regulation of angiogenesis, calcium ion binding and natural killer cell activation involved in immune response. With KEGG database analysis, down-regulated pathways included ECM-receptor interaction and peroxisome and up-regulated pathways included Glutathione metabolism and PPAR signaling pathway. The expressions of CD44 and IL-6R were significantly different and appeared biologically significant. CD44 was significantly upregulated in insensitive tissues (0.54±0.06) compared with sensitive tissues (0.33±0.04)(P<0.01). IL-6R was significantly downregulated in insensitive tissues (0.44±0.03) compared with sensitive tissues. (0.68±0.03) (P<0.01). Conclusion: CD44 and IL-6R may be potentially functional genes of TPF induction chemotherapy in hypopharyngeal squamous cell carcinoma.
目的： 应用基因表达谱芯片，筛选下咽鳞状细胞癌患者对紫杉醇+顺铂+5-氟尿嘧啶（TPF）方案诱导化疗敏感性的差异表达基因，并进行初步分析。 方法： 选取2013年1月至2017年12月北京同仁医院头颈外科29例原发下咽鳞状细胞癌患者（男28例，女1例，年龄43~73岁）TPF方案诱导化疗治疗前的新鲜肿瘤标本。分为诱导化疗敏感组（16例）和不敏感组（13例）。应用Illumina人类全基因组表达芯片检测2组的表达差异，对差异基因进行生物信息学分析。在29例样本基础上增加符合入组条件的14例样本，应用反转录实时定量聚合酶链反应（reverse transcription and quantitative real-time PCR，RT-qPCR）技术检测潜在功能核心基因在诱导化疗敏感组患者及诱导化疗不敏感组患者组织中表达的差异。采用Graphpad prism 7.0软件对测量结果进行统计分析。 结果： 共筛选出1 381个差异基因。经GO（Gene Ontology）数据库分析，功能上调的集合主要包括：细胞外基质隔离、趋化因子受体结合、钾离子通道调节活性等集合；下调的集合包括：血管生成调节、钙离子结合、自然杀伤细胞免疫应答活性等集合。经KEGG（Kyoto Encyclopedia of Genes and Genomes）数据库分析，下调通路主要有：细胞外基质受体相互作用通路、过氧化物酶体通路等；上调通路主要有：谷胱甘肽代谢通路、过氧化物酶体增殖物激活受体（peroxisome proliferator activated receptor，PPAR）信号通路等。其中CD44、IL-6R分别为表达下调基因和上调基因，可能位于核心功能调控位置。经RT-qPCR检测，CD44 mRNA在诱导化疗不敏感组织中相对表达量（0.54±0.06）显著高于诱导化疗敏感组织（0.33±0.04）中相对表达量（P<0.01）；IL-6R mRNA在诱导化疗不敏感组织中相对表达量（0.44±0.03）显著低于诱导化疗敏感组织中相对表达量（0.68±0.03），差异有统计学意义（P<0.01）。 结论： CD44、IL-6R可能是下咽鳞状细胞癌TPF诱导化疗敏感性相关的潜在功能性基因。.
Keywords: Bioinformatics; Carcinoma, squamous cell; Gene expression profiling; Hypopharyngeal neoplasms; Induced chemotherapy.