喉癌及喉咽癌颈部淋巴结微转移的检测及临床意义初探

目的 比较半连续切片H&E染色及免疫组化染色两种方法，用于检测喉及喉咽部鳞形细胞癌患者颈部常规病理学阴性淋巴结中微转移的检出率，并初步探讨其与临床病理因素及早期复发的关系。方法 对13例喉及喉咽部鳞型细胞癌患者经常规病理检查为阴性的15侧326个淋巴结, 采用50μm半连续切片苏木精－伊红染色法及细胞角蛋白CK(AE1/ AE3)免疫组化EnVison两步法行微转移灶检测, χ2 检验用于检验两种不同方法的阳性检出率。统计所有病例的各临床病理因素（年龄、性别、临床分型、pT 分期、肿瘤分级、原发灶浸润深度），随访11～56个月（中位时间31个月），采用χ2 检验分析各临床病理因素与微转移之间的相关性以及微转移与早期复发的相关性。结果 ①全部75个蜡块经半连续切片后免疫组化和苏木精－伊红染色切片各750张，免疫组化组中经CK(AE1/ AE3) 单克隆抗体标记后,共40张切片（5.33%）分别对应5 例(38.46 %) 患者中的7 个淋巴结(2.14 %)呈阳性反应。而相应的苏木精-伊红染色切片，若不与免疫组化对照，其阳性切片数为24张（3.20%），分别对应2例患者的3个淋巴结。两组间阳性切片数的检出率差异有统计学意义（Ρ<0.05）；②除年龄外的各临床病理因素与微转移之间差异无统计学意义( Ρ>0.05) ;有微转移和无微转移患者的早期复发率差异无统计学意义(Ρ>0.05) 。结论 对于喉及喉咽鳞癌的颈部淋巴结微转移，单纯的苏木精－伊红染色切片，即使是连续或半连续切片，有经验的病理科医师仍然会有一定的漏检率，而免疫组化方法可提高其检出率。微转移与临床病理因素及预后间的可能相关性尚有待大样本和长期随访资料的进一步探讨。

Detection of micrometastasis in cervical lymph nodes from laryngeal or laryngopharyngeal carcinoma and preliminary research on the clinical significance of micrometastasis

Objective To Compare the positive detection rates of hematoxylin and eosin stain with immunohistochemical stain by semi-consecutive sectioning of histopathologically negative cervical lymph nodes from laryngeal or laryngopharyngeal squamous cell carcinoma.The relationship between micrometastasis and clinicopathological factors, micrometastasis and early recurrences was studied primitively. Methods Totally, 326 negative cervical lymph nodes (CLNs) from 13 patients (15 sides), with laryngeal or laryngopharyngeal squamous cell carcinoma, were included in this study. Two methods( H&;E stain and EnVison′s two-step IHC technique with monoclonal antibody CK (AE1/ AE3)) combined with semi--consecutive sectioning(per-50μm) were applied to detect the micrometestasis . The positive detection rates of H&;E and IHC were compared by Chisquare test . The clinicopathological factors, consisted of sex, age, pT stage, clinical typing, grade of differentiation,depth of primary tumor invasion, were studied and all the cases were followed-up for 11～56 months（median time 31.4 months）. Further, the relationship between micrometastasis and clinicopathological factors, micrometastasis and early recurrences were analyzed with Chisquare test. Results 1. All the paraffin specimens of 75 cases were semi-consecutively sectioned into two sets of 750 slices for H&;E stain and IHC stain. In the IHC group, 40（5.33%）slices were found positive in 7 (2.14 %) lymph nodes which came from 5(38.46 %) patients Howerver, only 24（3.20%）positive slices were found in 3 lymph nodes of 2 patients in the H&;E group.The positive detection rates of these two groups were statistically significant（Ρ<0.05）. 2. There was no statistically significance between micrometastasis and the clinicopathological factors (Ρ>0.05) except for the age (Ρ=0.032). There was no statistically significance between micrometastasis and early recurrences. Conclusion 1. Even the experienced pathologists may miss the micrometastasis in cervical lymph nodes from laryngeal or laryngopharyngeal carcinoma if only by H&;E stain,though semi-serial or serial section was applied in those cases.While the IHC technique can obviously increace the positive detection rates. 2.The possible relationship between micrometastasis and clinicopathological factors, micrometastasis and recurrences should be verified with more samples and long-term follow-up.