结直肠癌周围淋巴结CK20蛋白检测的临床应用研究

Study on clinical application of colorectal carcinoma peripheral lymph node CK20 protein detection

目的:探讨免疫组化(IHC)检测结直肠癌区域淋巴结微转移的细胞角蛋白20(CK20)对结直肠癌术后临床分期、预后判断和治疗干预的可行性和临床应用价值。方法:选取40例常规HE病理检查淋巴结转移阴性的结直肠癌患者为研究对象,术前抽取外周血行循环肿瘤细胞(CTCs)测定;术中、后沿亚甲蓝染剂标识的区域淋巴结按4站次淋巴结分别摘除、放置并标记,应用IHC技术检测淋巴结中CK20,统计淋巴结微转移患者的阳性例数及阳性率及其在各不同站次的分布情况,并对HE、CTC和IHC CK20方法检测的结果进行比较。结果:淋巴结CK20测定与HE染色法的淋巴结微转移阳性检出率间差异有统计学意义(P<0.01);不同站次间淋巴结微转移阳性检出率存在统计学差异(P<0.01或P<0.05);全组CK20检测有存在淋巴结微转移11例(11/40,27.5%);7例患者TNM分期提高,HE染色法重新分期率为17.5%(7/40)。全组CTC检测显现阳性7例,阳性表达率为17.5%(7/40),其中6例对应于IHC淋巴结CK20(+)患者,1例为IHC淋巴结CK20(-)患者,且CTCs细胞学分型表达与结直肠癌肿瘤分化程度、肿瘤浸润深度呈正相关性。结论:CK20免疫组化法是检测淋巴结微转移可靠而敏感的方法,结合亚甲蓝染色法显影淋巴结可简便、快速地发现微转移灶,与CTC检测联用可达互补效能,进一步提高微转移检出率,减少假阴性发生。

Objective：To explore the feasibility and clinical application value of immunohistochemical （IHC） method detecting cytokeratin 20 （CK20） of colorectal carcinoma （CRC） lymph node micrometastasis in the postoperative clinical staging,prognostic diagnosis and therapeutic interventions for patients with CRC. Methods：A total of 40 patients with negative CRC lymph node metastasis by hemotoxylin and eosin HE pathological examination,and peripheral blood circulating tumor cells （CTCs） were collected preoperatively. The lymph nodes were incised,placed and marked every 4 locations along methylene blue stain-marked regional lymph nodes during and after operation,IHC was used to detect CK20 in lymph nodes,the number of patients with positive lymph node micrometastasis and its distribution in different stations were recorded, and the detection results of HE, CTC and IHC CK20 were compared. Results：There was significant differences in the positive detection rate of lymph node micrometastasis between lymph node CK20 and HE stain （ P 〈 0. 01 ） , and the differences were also significant in the positive detection rate of lymph node micrometastasis among different locations （P 〈 0.01 or P 〈 0. 05）. According to full-set CK20 detection, 11 （11/40,27.5%） patients had lymph node micrometastasis ;7 patients had increased TNM stages,and the re-staging rate of HE stain was 17.5% （7/40）, Full-set CTC detection showed that 7 patients were positive,with positive expression rate being 17.5% （7/40） ,in whom 6 were corresponding to IHC lymph node CK20（ ＋ ） while 1 to IHC lymph node CK20 （ - ）. Meanwhile, CTCs cytological typing expression was in positive association with CRC tumor differentiation severity and tumor infiltration depth. Conclusion：CK20 immunohistochemical method,as a reliable and sensitive method for the detection of lymph node micrometastasis,can simply and quickly find the micrometastatic nidi combing with methylene blue stain,and has synergistic effect with CTC detection to further increase the detection rate of lymph node micrometastasls and reduce the occurrence of false negative detection.