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1 材料和方法1.1 材料收集我院胸外科手术切除并经病理检查诊断为食管鳞癌的新鲜癌组织标本及远离肿瘤的正常食管组织作为对照各40例(未经放化疗及电化学治疗).其中男32例,女8例;年龄43岁～75岁(平均55.3岁).PTNM 分期参照1987年UICC 标准.其中,Ⅰ期0例;Ⅱ_A期10例;Ⅱ_B期9例;Ⅲ期19例,Ⅳ期2例.根据 p16基因组序列,在第2外显子两侧设计一对扩增引物;P1:5′-TGTTCYCTCTGGCAGGTCAT-3′;P2:5-CTCAGATCATCAGTCCTCAC-3′;PCR 产物长度342 bp.选用β-actin 做内对照,引物序列如下:P1:5′-GGCGGCACCACCAT-GTACCCT-3′;P2:5′-AGGGGCCGGACTCGTCATACT-3′,PCR 产物为312 bp.引物和4种10 mmol/L dNTP 混合液购自上海
1 Materials and Methods 1.1 Materials Collection Our hospital thoracic surgery resected and pathologically diagnosed as esophageal squamous cell carcinoma of fresh tissue specimens and normal esophageal tissue away from the tumor as control 40 cases (without radiotherapy and chemotherapy). There were 32 males and 8 females, ranging in age from 43 years to 75 years (mean 55.3 years) .PTNM staging was based on the UICC standard of 1987. Among them, 0 cases were in stage Ⅰ, 10 cases in stage Ⅱ_A, 9 cases in stage Ⅱ_B, 19 cases in stage Ⅲ , And 2 cases in stage Ⅳ.A pair of amplification primers was designed on both sides of exon 2 according to the sequence of p16 genome.P1: 5’-TGTTCYCTCTGGCAGGTCAT-3 ’; P2: 5-CTCAGATCATCAGTCCTCAC-3’ The sequence of the primers was as follows: P1: 5’-GGCGGCACCACCAT-GTACCCT-3 ’; P2: 5’-AGGGGCCGGACTCGTCATACT-3’, and the PCR product was 312 bp. The primers and four 10 mmol / L dNTPs Mixed liquid from Shanghai