抗菌肽hCAP18在结肠癌患者血清中的表达及其临床应用价值  

Detection and clinical application value of antimicrobial peptide hCAP18 in serum of colorectal carcinoma patients

作　　者：吴军录[1] 权文强[1] 姚懿雯[1] 张宇[1] 李泽兵[2] 李冬[1] 

机构地区：[1]同济大学附属同济医院检验科,上海200065 [2]上海市普陀区精神卫生中心检验科

出　　处：《中华检验医学杂志》2017年第4期278-283,共6页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金（81272603,81472179）;上海申康医院发展中心课题（SHDC22014008）

摘　　要：目的探讨结肠癌患者血清中抗菌肽人类阳离子抗菌蛋白18（hCAP18）水平的变化及其在结肠癌辅助诊断和预后中的价值。方法采用病例对照研究。选择同济大学附属同济医院胃肠外科2014年1月至2015年6月结肠癌患者标本68例及同期体检科40名健康对照者进行研究，用酶联免疫吸附试验（ELISA）检测血清中hCAP18的浓度，并比较结肠癌患者术前和术后血清中hCAP18的浓度变化情况；免疫组化检测结肠癌组织中的hCAP18的表达；采用溴脱氧核苷尿嘧啶（BrdU）——酶联免疫吸附试验（ELISA）法和软琼脂克隆形成实验检测hCAP18对结肠癌细胞增殖能力的影响。通过分析受试者工作特征曲线（ROC）判断hCAP18浓度在结肠癌诊断中的敏感度、特异度。采用t检验和单因素方差分析等方法进行统计学分析。 结果血清hCAP18水平在结肠癌Ⅰ期、Ⅱ期、Ⅲ期、Ⅳ期患者中分别为（0.46±0.18）mg/L、（0.65±0.45）mg/L、（1.26±0.68）mg/L、（2.35±1.06）mg/L，均值为（1.16±0.88）mg/L；显著高于健康对照组的（0.19±0.07）mg/L，差异有统计学意义（t＝5.290，P〈0.05）；在结肠癌患者肿瘤切除后30 d血清中hCAP18的浓度为（0.26±0.06）mg/L，明显低于术前均值（1.16±0.88）mg/L，差异有统计学意义（t＝3.971，P〈0.05）；免疫组化结果显示与癌旁组织相比，结肠癌组织中hCAP18高表达；BrdU-ELISA法检测的结果显示，用浓度为0.05 ～ 1.00 mg/L的hCAP18在处理后，HCT116和SW480细胞的增殖能力明显增加；软琼脂克隆形成实验显示结肠癌细胞HCT116和SW480经1.00 mg/L hCAP18处理后，HCT116细胞克隆簇大小由（145.40±35.20）μm增加到（370.80±32.65）μm，差异有统计学意义（t＝10.50，P〈0.05），数量分别由（8.50±2.30）个增加到（42.80±6.60）个，差异有统计学意义（t＝3.945，P〈0.05）；SW480细胞克隆簇的大小由（101.00±27.10）μm增加到（36ObjectveTo evaluate the serum level of antimicrobial peptide human cationic antimicrobial protein 18 （hCAP18） in colorectal patients and it auxiliary diagnosis and prognosis value.MethodsCase-control study was used. The serum level of hCAP18 was measured by enzyme linked immunosorbent assay（ELISA） in 68 cases with colorectal patients of department of gastrointestinal surgery and 40 cases healthy people of department of physical examination from January 2014 to June 2015 in Tongji Hosptial of Tongji University. The concentrations of hCAP18 in serum of colorectal patients before and surgery were analyzed. Immunohistochemistry was used to detect hCAP18 expression in colorectal carcinoma. The effect of hCAP18 on colon carcinoma cell proliferation was detected by BrdU-ELISA and soft agar colony formation assay. The sensitivity and specificity of serum hCAP18 for the diagnosis of colorectal were evaluated using the receiver operating characteristic curves（ROC） . Date was analyzed by using the t-test and one-way analysis of variance.ResultshCAP18 serum levels in colon cancer of stage Ⅰ, Ⅱ, Ⅲ and IV patients were （0.46±0.18）mg/L, （0.65±0.45）mg/L, （1.26±0.68）mg/L and （2.35±1.06）mg/L. Mean value was（1.16±0.88） mg/L, which was significantly higher than in normal people （0.19±0.07）mg/L（t=5.290, P〈0.05）. hCAP18 levels had significantly decreased in serum of colorectal patients after 30 d surgery compared to preoperative results[from （1.16±0.88）mg/L to（0.26±0.06）mg/L; t=3.971, P〈0.05]. Immunohistochemistry results showed hCAP18 was high expression in colon cancer tissue compared with adjacent tissues; BrdU-ELISA assay results showed HCT116 and SW480 cell proliferation increased significantly after 0.05-1 mg/L of hCAP18 treatment; Soft agar clone formation experiment proved hCAP18 could significant enhance clone formation of HCT116 and SW480 colon cancer cell lines. The size of clonal cluster of HCT116 was increased from （145.40±35.20）μm to（370.80±32.65�

关 键 词：抗微生物阳离子肽类 肿瘤标记 生物学 结肠肿瘤 细胞增殖 

分 类 号：R730.43[医药卫生—肿瘤] R735.35[医药卫生—临床医学]