摘要目的 分析Kruppel样因子4(KLF4)和CD44在食管鳞状细胞癌组织和对应的癌旁组织中的表达情况,并探讨其对预后的影响.方法 选取2012年6月至2016年9月于南阳市第二人民医院外科行手术治疗的100例食管鳞状细胞癌患者,取其食管鳞状细胞癌组织及对应的癌旁正常组织(对照),采用免疫组织化学法检测其K L F4和CD44的蛋白质表达情况;选取其中50例患者的新鲜组织标本,采用实时荧光定量PCR法和蛋白质印迹法分别检测KLF4、CD44的mRNA与蛋白质表达水平.统计学分析采用t检验、卡方检验、Kaplan-Meier法和Pearson相关性分析.结果 KLF4蛋白质在食管鳞状细胞癌组织中的阳性率为55％(55/100),低于对应癌旁组织的77％(77/100),差异有统计学意义(χ2=10.778,P=0.001).CD44蛋白质在食管鳞状细胞癌组织中的阳性率为81％(81/100),高于所对应癌旁组织的11％(11/100),差异有统计学意义(χ2=112.600,P<0.01).食管鳞状细胞癌组织中K L F4的mRNA表达水平比癌旁组织降低者43例,CD44的mRNA表达水平比癌旁组织升高者46例,差异均有统计学意义(χ2=51.837、70.563,P均<0.01).癌组织中K L F4的阳性表达在不同性别、不同分化程度、不同浸润深度、不同TNM分期和有无淋巴结转移中差异均有统计学意义(P均<0.05).癌组织中CD44的阳性表达在不同分化程度、不同浸润深度、不同T N M分期和有无淋巴结转移中差异均有统计学意义(P均<0.05).不管是蛋白质水平还是mRNA水平,KLF4与CD44的表达均呈负相关(r=-0.284、-0.518,P均<0.01).食管鳞状细胞癌组织中K L F4蛋白质阳性患者的中位生存期为33个月,长于K L F4蛋白质阴性患者的20个月,差异有统计学意义(χ2=4.021,P=0.019);食管鳞状细胞癌组织中CD44蛋白质阳性患者的中位生存期为24个月,短于CD44蛋白质阴性患者的37个月,差异有统计学意义(χ2=4.379,P=0.016).单因素分析显示,T N M分期、K L F4表达和CD44表达与总生存期相关(P均<0.05);多因素分析显示,TNM分期、KLF4低表达和CD44高表达是影响患者生存的独立危险因素(P均<0.05).结论 食管鳞状细胞癌组织中的K L F4低表达和CD44高表达可能与其发生、发展和预后密切相关.

abstractsObjective To analyze the expression of Kruppel-like factor 4 (KLF4 ) and CD44 in esophageal squamous cell carcinoma (ESCC) tissues and adjacent non-cancerous tissues ,and to investigate their effects on the prognosis .Methods From June 2012 to September 2016 ,in The Second People′s Hospital of Nanyang ,a total of 100 patients with ESCC who receiving operation were selected .The ESCC tissues and the adjacent non-cancerous tissues (control) of the patients were collected .The expression levels of KLF4 and CD44 protein were detected by immunohistochemistry .The expressions of KLF4 and CD44 at mRNA and protein level of 50 paired fresh tissues were examined by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting ,respectively . T-test ,chi-square ,Kaplan-Meier method and Pearson correlation analysis were performed for statistical analysis .Results The positive expression rate of KLF4 protein in ESCC tissues was 55％ (55/100) ,which was lower than that in adjacent non-cancerous tissues (77％ ,77/100) ,and the difference was statistically significant (χ2 =10 .778 ,P=0 .001) .The positive expression rate of CD44 protein in ESCC tissues was 81％ (81/100) ,which was higher than that in adjacent non-cancerous tissues (11％ ,11/100) ,and the difference was statistically significant (χ2=112 .600 ,P<0 .01) .The expression level of KLF4 mRNA in 43 cases was lower than that in adjacent non-cancerous tissues ,the expression level of CD44 mRNA in 46 cases was much higher than that of adjacent non-cancerous tissues ,and the differences were statistically significant (χ2 =51 .837 and 70 .563 ,both P< 0 .01) .There were statistically significant differences in positive expression rates of KLF4 in cancer tissues between different gender , differentiation degree , invasion depth ,TNM stage and lymph node metastasis (all P<0 .05) .Similarly there were statistically significant differences in positive expression rates of CD 44 in cancer tissues between different differentiation degree ,invasion depth ,TNM stage and lymph node metastasis (all P< 0 .05) .The expression of KLF4 was negatively correlated with CD44 expression ,either at protein level or mRNA level (r= -0 .284、-0 .518 ,both P< 0 .01) .The median survival time of patients with positive KLF4 expression in cancer tissues was 33 months ,which was longer than that of patients with negative KLF4 expression (20 months) ,and the difference was statistically significant (χ2 =4 .021 , P= 0 .019) .The median survival time of patients with positive CD44 expression in cancer tissues was 24 months ,which was shorter than that of patients with negative CD44 expression (37 months) , and the difference was statistically significant (χ2 = 4 .379 , P= 0 .016) .The results of univariate analysis showed that TNM stage ,KLF4 expression and CD44 expression were correlated with overall survival time (all P<0 .05) . The results of multivariate analysis indicated that TNM stage ,lower KLF4 expression and higher CD44 expression were the independent risk factors for survival (all P<0 .05) .Conclusion Lower expression of KLF4 and higher expression of CD44 in ESCC may be closely correlated with its occurrence ,development and prognosis .