| Abstract: | 研究目的︰原發子宮頸腺癌與原發子宮內膜腺癌因病變解剖位置靠近，有時在臨床上與病理上，都不易區分；尤其發生上下延伸侵犯時，更難以區分何者為原發部位。二者基本生物特性差異極大，臨床上這兩個地方癌症的治療方式完全不同，而且一直缺乏可靠的區分方式。當發生腺癌來源無法判斷時，在婦科病理學中，免疫組織染色法可提供輔助診斷。本研究包括三部份﹕(1)找出最適當且簡單的計分法可以應用於p16INK4a免疫組織染色，在臨床上輔助鑑別診斷子宮頸腺癌及子宮內膜腺癌。(2)除p16INK4a外，再針對其他常用於鑑別診斷子宮頸腺癌及子宮內膜腺癌的四種組織免疫標誌（包括ER, PR, Vim, CEA），進行驗證﹔(3)期望能整合數種潛在有用的免疫組織標誌，開發出最妥適生物標誌診斷套組，以進行子宮頸腺癌與子宮內膜腺癌鑑別診斷。
材料與方法︰本研究樣本來自早年就已經去連結檢體，包括14例子宮頸腺癌與21例子宮內膜腺癌，製成石蠟包埋之組織蠟塊，做成微陣列排序的組織晶片。以五種生物標誌(包括ER, PR, Vim, CEA, p16INK4a)抗體，採用avidin-biotin complex 技術，進行免疫組織染色。採用semiquantative計分法，考量染色強度及區城面積做評分的依據。
結果︰我們的結果顯示(1) 在p16INK4a的四個計分方法中，包括「單獨胞核評估計分法」、「單獨胞質評估計分法」、「胞核與胞質分數之平均計分法」、「胞核與胞質之單獨計分較高者計分法」等四者；其中之p16INK4a免疫組織呈色的「單獨胞質評估計分法」，對區分子宮頸腺癌及子宮內膜腺癌並不具統計學上意義，而其他三種計分法則有明顯差異。(2) 除p16INK4a以外，其餘四種生物標誌（ER, PR, Vim, CEA），在子宮頸腺癌及子宮內膜腺癌的免疫組織呈色，也都出現有意義的顯著差異。(3)經比較各種組合，我們證實︰以二合一生物標誌 (Vim，CEA) 組合，在鑑別原發性子宮頸腺癌及子宮內膜腺癌的診斷中，呈現最佳的整體準確度(overall accuracy)，達0.783 (0.648, 0.917)。
結論: 我們的研究證實︰(1) 使用p16INK4a生物標誌時，以較為簡單方便的單獨胞核評估計分法，不需考慮細胞質呈色的結果，就足以區分原發性子宮頸腺癌與原發性子宮內膜腺癌。(2)另外四種個別生物標誌（ER, PR, Vim, CEA），對區分原發性子宮頸腺癌與原發性子宮內膜腺癌，亦有幫助。(3) 五合一、四合一、三合一、及二合一的各種生物標誌組合(panels)，都可以用於鑑別診斷原發性子宮頸腺癌與原發性子宮內膜腺癌， 但二合一生物標誌組合中之 Vim/CEA panel， 應該是最經濟有效的組合了。
Objective: The choice of appropriate therapeutic plans for primary endocervical adenocarcinomas (ECA) and endometrial adenocarcinomas (EMA) depends on the tumor’s site of origin, because they have differing biological behaviors. Although careful morphologic examination usually results in a confident diagnosis, difficulties may also arise in the pathological examination of a large surgically resected specimen for a tumor that involves both the lower uterine segment and upper endocervix. When there is doubt and indistinguishable primaries, immunohistochemistry (IHC) may assist. The purposes of this study are: (1) to define a sufficient, simple and useful scoring mean of p16INK4a in distinguishing between ECA and EMA (2) to compare the individual expression status of five immunomarkers (ER, PR, Vim, CEA and p16INK4a). (3) to determine the most favorable panel in distinguishing between ECA and EMA.
Methods and materials: A tissue microarray (TMA) was constructed using paraffin-embedded, formalin-fixed tissues from 35 hysterectomy specimens, including 14 ECA and 21 EMA. TMA sections were immunostained with five antibodies, by avidine-biotin complex (ABC) method for antigen visualization. The staining intensity and area extent of the immunohistochemical (IHC) reactions were appraised by using the semi-quantitative scoring system.
Result: (1) Method Cytoplasm for p16INK4a IHC was not a statistically significant method to distinguish between ECA and EMA. However the other three methods for p16INK4a IHC, Method Nucleus, Method Mean of Cytoplasm plus Nucleus, and Method Dominant Cytoplasm or Nucleus, resulted in significant frequency differences between ECA and EMA. The overall accuracy rate of Method Mean of Cytoplasm plus Nucleus was 80 %, the highest among the four scoring methods. (2) The expressions of five respective markers (ER, PR, Vim, CEA and p16INK4a) showed significant frequency differences between ECA and EMA. (3) The 2-markers (Vim, CEA) panel exhibited the most efficiency in the diagnostic distinction between primary ECA and EMA.
Conclusions: Method Nucleus, based on independent of cytoplasmic stains, can conveniently and effectively distinguish between ECA and EMA. The 2- markers (Vim, CEA) panel would be sufficient in making a diagnostic distinction between primary ECA and EMA |
