肝外胆管癌淋巴结微转移的检测及其对预后的影响

Detection of lymph node micrometastasis for patients with extrahepatic cholangiocarcinoma and its prognostic significance

目的观察CK19和CK20在肝外胆管癌（EHCC）淋巴结微转移中的表达，探讨淋巴结微转移与临床病理特征和CA19-9、癌胚抗原（CEA）的关系，以及淋巴结微转移对预后的影响。方法选取59例行手术切除EHCC患者的279枚淋巴结，分别以CK19、CK20单克隆抗体进行免疫组化染色和常规HE染色，确定有无淋巴结转移与微转移，并与各病例的临床病理及随访资料进行比较，分析微转移与临床病理因素的关系及对生存率的影响。结果59例患者的淋巴结转移率，HE诊断仅23．72％（14／59），而CK诊断为35．59％（21／59，P〈0．05）；淋巴结转移发生率由HE染色的5．37％提高到CK染色的13．98％（P〈0．05）；在常规HE染色诊断没有淋巴结转移的45例患者中，微转移的发生率为15．56％（7／45）。淋巴结微转移术前血清CA19-9浓度显著高于无淋巴结微转移者（对照组）（P〈0．05）；淋巴结微转移与血清CA19-9浓度呈正相关（rs=0．371，P〈0．05）。对12个因素进行Logistic回归分析，结果显示肿瘤的组织学类型和淋巴浸润对淋巴结微转移有重要影响（P〈0．05）。结论CK免疫组化染色能检测出HE染色阴性淋巴结中隐匿的癌细胞，淋巴结微转移能够更准确地判断EHCC患者的预后。

Objective To observe the expression of antibodies of cytokeratin 19 and 20 in lymph node micrometastasis in patients with extrahepatic cholangiocareinoma （ EHCC ）, evaluate the prognostic significance of lymph node （LN） micrometastasis and study the correlation between lymph node micrometastasis and clinicopathological features, CA19-9 and CEA. Methods A total of 279 lymph nodes was intra-operatively collected from 59 EHCC patients and routine histological examination performed. Immnnohistochemical staining was performed on all samples by the murine antibodies of anti-CK19 and anti- CK20 respectively. Then the micrometastasis was identified microscopically according to the color of cells. The results were analyzed according to clinical, pathological and follow-up data. And the relation of micrometastasis with clinical pathological factors and its impact upon survival rate were analyzed. Results Among59 EHCC patients, 14 （23.72%） LN metastasis were found with HE staining and 21 micrometastases with CK staining. The incidence of nodal involvement in 59 EHCC patients increased from 5.37% （15/279） by HE staining to 13.98% （39/279） by CK staining. Among 45 patients not positive for LN metastases with HE staining, CK staining was positive in 7 patients and the incidence of mierometastasis was 15.56%. The preoperative serum CA19-9 levels in patients with LN micrometastasis was higher than that those without LN metastasis （P 〈 0. 05 ）. And there was a positive correlation between occult nodal micrometastasis and serum concentrations of CA19-9 （ r5 = 0. 371, P 〈 0. 05）. The histological type and lymphatic vessel infiltration of tumor were the most importance factors for LN micrometastasis through Logistic regression analysis （ P 〈 0. 05 ） . Conclusion The CK immunohistochemical staining can detect the micrometastases in HE negative LN. And LN micrometastasis can more accurately predict the prognosis of EHCC patients.