表皮生長(zhǎng)因子及其受體在食管癌中的表達(dá)

1、表皮生長(zhǎng)因子及其受體在食管癌中的表達(dá)【摘要】目的研究表皮生長(zhǎng)因子(epidermal growth factor, EGF)及其受體(EGF receptor, EGFR)在食管鱗狀細(xì)胞癌中的表達(dá)。方法用免疫組織化學(xué)ABC法分析并研究染色結(jié)果與病理改變之間的關(guān)系。EGF染色，以正常人頜下腺作為陽(yáng)性對(duì)照。EGFR以正常人胎盤組織為陽(yáng)性對(duì)照。EGF和EGFR染色一抗的濃度分別為1200，1100，4孵育48h。結(jié)果EGF強(qiáng)陽(yáng)性18例，弱陽(yáng)性16例，EGFR強(qiáng)陽(yáng)性18例，弱陽(yáng)性16例。EGF和EGFR的表達(dá)分為兩種類型：M型：癌組織本身免疫組織化學(xué)反應(yīng)強(qiáng)弱不同，D型：癌組織中免疫組織化學(xué)反應(yīng)強(qiáng)度均一

2、。D型與M型在EGF、EGFR的表達(dá)中較一致。M型較多發(fā)生淋巴結(jié)轉(zhuǎn)移(P0.05)，并且強(qiáng)陽(yáng)性組中M型所占比例較大。結(jié)論EGF及EGFR強(qiáng)陽(yáng)性表達(dá)同食管癌病人的預(yù)后有關(guān)?！娟P(guān)鍵詞】表皮生長(zhǎng)因子(EGF)表皮生長(zhǎng)因子受體(EGFR)食管癌免疫組織化學(xué)EXPRESSION OF EPIDERMAL GROWTH FACTOR ANDITS RECEPTOR IN ESOPHAGEAL CANCERDing Mingjie，Ding Wei，Yin Deren*，Bai Jingxiu，Zhang Wei，Gao Xiaodong，Zhang Linqing，Niu Fuwen，F(xiàn)u Shixian

3、(Department of Histology and Embryology, Henan Medical University, Zhengzhou,*Department of Pathology, Xinxiang Central Hospital)【Abstract】 Objective In order to detect the expression of both EGF and EGFR in human esophageal squamous cell carcinoma. Method The specimens from 34 patients with esophag

4、eal cancer were studied by immunohistochemical assay. Results The EGF and EGFR were faintly expressed in basal and parabasal layers of normal esophageal epithelia. No correlation was found between the intensity of EGF、EGFR staining and the histologic type, the patients with strongly expressed EGF an

5、d EGFR had lymph node metastases more frequently. The EGF and EGFR expression showed a mosaic (M) pattern in 15 patients and diffuse (D) pattern in 19 patients. The patients with a M pattern had lymph node metastases more frequently than those with a D pattern. In cancer cells, EGFR was stained both

6、 in the cytoplasm and on the cell membrane, some of them in the nucleus. Conclusion The overexpression of EGF and EGFR could be of prognostic significance in esophageal cancer.【Key words】 Epidermal growth factor(EGF); Epidermal growth factor receptor (EGFR); Esophageal carcinoma; Immunohistochemistr

7、yEGF是美國(guó)學(xué)者Cohen1首先發(fā)現(xiàn)的。近年來(lái)發(fā)現(xiàn)，erb-B癌基因產(chǎn)物與EGF受體十分相似。已有報(bào)道，在肺癌，乳腺癌，胃癌，膀胱癌，腦瘤，胰腺癌等2腫瘤組織中，含有EGF或EGF受體，并發(fā)現(xiàn)EGF受體同腫瘤病人的預(yù)后有關(guān)3。EGF能刺激某些腫瘤細(xì)胞株的生長(zhǎng)4，5。可見，EGF在腫瘤的發(fā)生發(fā)展過(guò)程中起重要的作用。有關(guān)表皮生長(zhǎng)因子同食管癌發(fā)生的關(guān)系方面，近來(lái)國(guó)外已有少數(shù)報(bào)道，且報(bào)道結(jié)果不一6，國(guó)內(nèi)尚未見報(bào)道。為進(jìn)一步探討表皮生長(zhǎng)因子同食管癌發(fā)生、發(fā)展的關(guān)系。我們運(yùn)用免疫組織化學(xué)方法，檢測(cè)食管癌組織中EGF及EGF受體的表達(dá)情況，結(jié)果發(fā)現(xiàn)，34例食管癌病例中EGF和EGFR均有不同程度的表達(dá)，E

8、GF強(qiáng)陽(yáng)性組18例，EGFR強(qiáng)陽(yáng)性組18例，EGF和EGFR的表達(dá)分為M型和D型，M型較多發(fā)生淋巴結(jié)轉(zhuǎn)移(P0.05(2檢驗(yàn))，顯示差異無(wú)顯著性(表1)。表1表皮生長(zhǎng)因子與食管癌分級(jí)的關(guān)系Table 1 Correlation between EGF and esophageal squamous cell carcinoma(SCC) grade食管癌分級(jí)grade of SCC例數(shù)casesEGF免疫反應(yīng)強(qiáng)度intensity of EGF staining強(qiáng)陽(yáng)性弱陽(yáng)性+(+)126612841046合計(jì)341816 ：高分化鱗癌 well differentiated squamous

9、cell carcinoma(SCC):中分化鱗癌 moderately differentiated squamous cell carcinoma(SCC)：低分化鱗癌 poorly differentiated squamous cell carcinoma(SCC)EGF：表皮生長(zhǎng)因子epidermal growth factor. 2=1.62 P0.051.2表皮生長(zhǎng)因子同食管癌淋巴結(jié)轉(zhuǎn)移的關(guān)系：34例食管癌中，發(fā)生淋巴結(jié)轉(zhuǎn)移的有17例。EGF樣反應(yīng)強(qiáng)陽(yáng)性組中有14例發(fā)生淋巴結(jié)轉(zhuǎn)移(2)，EGF弱陽(yáng)性組中3例有淋巴結(jié)轉(zhuǎn)移。而在未發(fā)生淋巴結(jié)轉(zhuǎn)移的病例中，強(qiáng)陽(yáng)性組僅有4例，弱陽(yáng)性組有1

10、3例(35)。同時(shí)在轉(zhuǎn)移的淋巴結(jié)中檢測(cè)到較強(qiáng)的EGF反應(yīng)(6)。說(shuō)明反應(yīng)強(qiáng)陽(yáng)性組發(fā)生淋巴結(jié)轉(zhuǎn)移的機(jī)率明顯大于弱陽(yáng)性組(P0.01)(表2)。1.3EGF陽(yáng)性反應(yīng)強(qiáng)度與陽(yáng)性反應(yīng)類型的關(guān)系：34例食管癌中，反應(yīng)類型為M型和D型者各為15例和19例(2，3)。M型在強(qiáng)陽(yáng)性組為11例，在弱陽(yáng)性組為4例。D型在強(qiáng)陽(yáng)性組和弱陽(yáng)性組分別為7例或12例。強(qiáng)陽(yáng)性組中M型多于D型(P0.05)(表3)。表2表皮生長(zhǎng)因子與食管癌淋巴結(jié)轉(zhuǎn)移的關(guān)系Table 2 Correlation between EGF and lymph node metastasisEGF陽(yáng)性分組groups of EGF staining淋

11、巴結(jié)轉(zhuǎn)移lymph node metastasisn(+)n(-)強(qiáng)陽(yáng)性組(+)strongly expressed144弱陽(yáng)性組(+)faintly expressed313合計(jì)1717 EGF:表皮生長(zhǎng)因子 epidermal growth factorn(+)：有淋巴結(jié)轉(zhuǎn)移 lymph node metastasisn(-):無(wú)淋巴結(jié)轉(zhuǎn)移 no lymph node metastasis 2=11.81 P0.01表3表皮生長(zhǎng)因子陽(yáng)性反應(yīng)強(qiáng)度同陽(yáng)性反應(yīng)類型的關(guān)系Table 3 Correlation between EGF and staining patternEGF陽(yáng)性分組group

12、s of EGF stainingEGF陽(yáng)性反應(yīng)類型pattern of EGF-IRM型D型強(qiáng)陽(yáng)性組(+)(18例)strongly expressed117弱陽(yáng)性組(+)(16例)faintly expressed412 EGF：表皮生長(zhǎng)因子 epidermal growth factorM型：不均一型 mosaic patternD型：均一型 diffuse pattern 2=4.48 P0.05)。2.2EGFR陽(yáng)性反應(yīng)強(qiáng)度同食管癌淋巴結(jié)轉(zhuǎn)移的關(guān)系：共有17例發(fā)生淋巴結(jié)轉(zhuǎn)移，其中EGFR強(qiáng)陽(yáng)性的有12例，EGFR弱陽(yáng)性的有5例。在未發(fā)生淋巴結(jié)轉(zhuǎn)移的病例中，EGFR免疫反應(yīng)強(qiáng)陽(yáng)性組中有

13、6例未發(fā)生淋巴結(jié)轉(zhuǎn)移，弱陽(yáng)性組中11例未發(fā)生轉(zhuǎn)移。在有鱗癌轉(zhuǎn)移的淋巴結(jié)中同樣檢測(cè)出有較強(qiáng)的EGFR免疫反應(yīng)(11)，即EGFR強(qiáng)陽(yáng)性組比EGFR弱陽(yáng)性組更容易發(fā)生淋巴結(jié)轉(zhuǎn)移。(P0.05)(表4)。表4表皮生長(zhǎng)因子受體同食管癌淋巴結(jié)轉(zhuǎn)移的關(guān)系Table 4 Correlation between EGFR and lymph node metastasisEGFR免疫反應(yīng)分組groups of EGFR-IR淋巴結(jié)轉(zhuǎn)移lymph node metastasesn(+)n(-)強(qiáng)陽(yáng)性組(18例)(+)strongly expressed126弱陽(yáng)性組(16例)(+)faintly express

14、ed511 n(+):有淋巴結(jié)轉(zhuǎn)移 lymph nodes metastasisn(-)：無(wú)淋巴結(jié)轉(zhuǎn)移 no lymph nodes metastasis 2=4.25 P0.05)。2.4表皮生長(zhǎng)因子(受體)陽(yáng)性反應(yīng)類型同淋巴結(jié)轉(zhuǎn)移的關(guān)系：17例淋巴結(jié)轉(zhuǎn)移的病例中M型為12例，D型為3例。在未發(fā)生淋巴結(jié)轉(zhuǎn)移的病例中M型為3例，D型為14例，在發(fā)生淋巴結(jié)轉(zhuǎn)移的病例中，表皮生長(zhǎng)因子(受體)免疫反應(yīng)類型為M型者明顯多于D型(P0.01)(表5)。表5表皮生長(zhǎng)因子陽(yáng)性反應(yīng)類型同淋巴結(jié)轉(zhuǎn)移的關(guān)系Table 5 Correlation between EGFR-IR pattern and lymph

15、node metastasis表皮生長(zhǎng)因子陽(yáng)性反應(yīng)類型pattern of EGF staining淋巴結(jié)轉(zhuǎn)移lymph node metastasesn(+)n(-)M型(15例)123D型(19例)514 n(+):有淋巴結(jié)轉(zhuǎn)移lymph nodes metastasisn(-)：無(wú)淋巴結(jié)轉(zhuǎn)移no lymph nodes metastasisM型：不均一型 mosaic pattern D型：均一型 diffuse pattern 2=9.66 P0.01討論表皮生長(zhǎng)因子及其受體與食管癌EGF通過(guò)與EGF受體結(jié)合，在細(xì)胞內(nèi)傳遞中構(gòu)成一個(gè)復(fù)雜的代謝網(wǎng)絡(luò)，控制著細(xì)胞的代謝、生長(zhǎng)、分化與癌變。在

16、實(shí)驗(yàn)中，34例食管癌組織的EGF和EGFR均有不同程度的表達(dá)。說(shuō)明表皮生長(zhǎng)因子同食管癌的關(guān)系密切。就其分布的部位來(lái)看，EGF和EGFR在食管癌細(xì)胞的胞膜、胞漿及胞核中均有表達(dá)。根據(jù)表皮生長(zhǎng)因子與其受體的作用機(jī)制，可以認(rèn)為，檢測(cè)到的EGF或EGFR，可能就是被內(nèi)轉(zhuǎn)入胞漿的EGF與其受體的復(fù)合物。本實(shí)驗(yàn)未能證實(shí)結(jié)合的EGF是外源性的，還是細(xì)胞自身的分泌物。已有報(bào)道，在鱗癌的發(fā)生中存在有自分泌機(jī)制(autocrine system)。7關(guān)于EGF受體在核中分布有不少報(bào)道，用I125標(biāo)記的mEGF為探針，在肝細(xì)胞核發(fā)現(xiàn)了EGF的結(jié)合點(diǎn)8。但食管癌細(xì)胞核中存在EGFR還未見報(bào)道。這可能同DNA的合成有關(guān)

17、9，10。表皮生長(zhǎng)因子可以刺激c-fos,c-myc等原癌基因的轉(zhuǎn)錄與表達(dá)，已發(fā)現(xiàn)在食管細(xì)胞中c-myc基因的擴(kuò)增，它們多為細(xì)胞核上的DNA結(jié)合蛋白，可以調(diào)節(jié)其他基因的表達(dá)。在實(shí)驗(yàn)中，正常上皮細(xì)胞核中沒(méi)有發(fā)現(xiàn)有EGFR及EGF樣免疫反應(yīng)，而在某些食管癌細(xì)胞中，卻發(fā)現(xiàn)有EGFR及EGF的表達(dá)。因此，我們可以認(rèn)為，存在于細(xì)胞核中的EGFR同細(xì)胞的癌變有某種關(guān)系。食管癌組織發(fā)生淋巴結(jié)轉(zhuǎn)移的機(jī)率較大。在本實(shí)驗(yàn)中，34例食管癌發(fā)生淋巴轉(zhuǎn)移者有17例(占50%)。無(wú)論是EGF還是EGFR，強(qiáng)陽(yáng)性組發(fā)生淋巴結(jié)轉(zhuǎn)移的明顯多于弱陽(yáng)性組。概率分別為P0.01和P0.05。另外，在免疫反應(yīng)性的類型上，陽(yáng)性反應(yīng)表現(xiàn)為

18、不均一型(M型)的比表現(xiàn)為均一型(D型)的較多地發(fā)生淋巴結(jié)轉(zhuǎn)移P0.01，這可能是表現(xiàn)為M型的食管癌組織中EGFR的濃度不同所致。眾所周知，淋巴結(jié)轉(zhuǎn)移和腫瘤侵潤(rùn)深度是判定食管癌預(yù)后的重要因素。但因我們尚未進(jìn)行臨床追蹤，我們只能初步認(rèn)為，同時(shí)表達(dá)EGF及EGFR強(qiáng)陽(yáng)性，以及反應(yīng)類型為不均一型(M型)同食管癌病人的預(yù)后有關(guān)。我們還發(fā)現(xiàn)在正常食管(斷端正常組織)上皮的基底層及基底旁層細(xì)胞表皮生長(zhǎng)因子及受體呈弱陽(yáng)性反應(yīng)，食管腺體未發(fā)現(xiàn)陽(yáng)性反應(yīng)，食管腺導(dǎo)管呈陽(yáng)性反應(yīng)，上皮的中間層相當(dāng)于棘細(xì)胞層的部位卻出現(xiàn)較強(qiáng)的陽(yáng)性反應(yīng)。Sato等提出，食管癌細(xì)胞可能起源于基底細(xì)胞和/或基底旁層細(xì)胞。本實(shí)驗(yàn)支持這種理論，

19、因?yàn)樵谡Ｇ闆r下，這些細(xì)胞具有較少的EGFR和EGF，而癌變的細(xì)胞卻具有較多的EGFR和EGF。EGFR基因的擴(kuò)增可能是其癌變的機(jī)制之一9。因?yàn)橐寻l(fā)現(xiàn)多種癌基因產(chǎn)物同EGFR具有高度同源性，另外，腫瘤細(xì)胞還能分泌一些生長(zhǎng)因子，這些生長(zhǎng)因子同EGF具有明顯的免疫交叉反應(yīng)。從而使EGFR持續(xù)受到刺激，使細(xì)胞發(fā)生轉(zhuǎn)化。從某種意義上講，細(xì)胞的癌變是以某種方式干擾或破壞生長(zhǎng)因子的正常信號(hào)通路，使細(xì)胞的生長(zhǎng)、分裂和分化從有控狀態(tài)變?yōu)槭Э貭顟B(tài)。至于棘細(xì)胞層的較強(qiáng)的陽(yáng)性反應(yīng)，可能是維持其本身細(xì)胞生長(zhǎng)、分化所必須的。因而，EGF是誘導(dǎo)腫瘤的發(fā)生還是促進(jìn)正常細(xì)胞生長(zhǎng)，可能取決于作用的細(xì)胞的類型、內(nèi)外環(huán)境以及EGF

20、的濃度等因素。版說(shuō)明1正常人頜下腺，ABC法，EGF樣免疫反應(yīng)性 2002ABC法，EGF樣免疫反應(yīng)染色強(qiáng)陽(yáng)性不均一型，(M型)。食管鱗癌級(jí)，有淋巴結(jié)轉(zhuǎn)移 2003ABC法，EGF樣免疫反應(yīng)弱陽(yáng)性，均一型(D型)。食管鱗癌級(jí)，無(wú)淋巴結(jié)轉(zhuǎn)移 2004ABC法，EGF樣免疫反應(yīng)強(qiáng)陽(yáng)性，均一型(D型)。食管鱗癌級(jí)，有淋巴結(jié)轉(zhuǎn)移 2005ABC法，EGF樣免疫反應(yīng)弱陽(yáng)性，(M型)。食管鱗癌級(jí)，無(wú)淋巴結(jié)轉(zhuǎn)移 2006ABC法，EGF樣免疫反應(yīng)強(qiáng)陽(yáng)性，(M型)。有轉(zhuǎn)移的淋巴結(jié) 1007ABC法，EGFR樣免疫反應(yīng)性。正常人胎盤組織 2008ABC法，EGFR樣免疫反應(yīng)強(qiáng)陽(yáng)性，(M型)。食管鱗癌級(jí)，有淋巴結(jié)

21、轉(zhuǎn)移 4009ABC法，EGFR樣免疫反應(yīng)弱陽(yáng)性，(D型)。食管鱗癌級(jí)，未發(fā)生淋巴結(jié)轉(zhuǎn)移 20010ABC法，EGFR樣免疫反應(yīng)性?？梢妭€(gè)別細(xì)胞核中出現(xiàn)陽(yáng)性反應(yīng)顆粒 40011ABC法，EGFR樣免疫反應(yīng)性。轉(zhuǎn)移的淋巴結(jié)，(M型) 10012ABC法，EGFR樣免疫反應(yīng)性。正常食管上皮 200Explanation of figuresFig.1 EGF-IR in normal human submandibular gland, ABC method 200Fig.2 Strongly expressed EGF-IR appeared as mosaic (M) pattern in h

22、uman esophageal squamous cell carcinoma(grade ) with lymph node metastasis. ABC method 200Fig.3 Faintly expressed EGF-IR appeared as a diffuse (D) partten in human esophageal squamous cell carcianoma (grade ) without lymph node metastasis. ABC method 200Fig.4 Strongly expressed EGF-IR appeared as a

23、diffuse (D) pattern in human esohpageal squamous cell carcinoma (grade) with lymph node metastasis. ABC method 200Fig.5 Faintly expressed EGF-IR appeared as a masaic (M) pattern in human esophageal squamous cell carcinoma (grade )without lymph node metastasis. ABC method 200Fig.6 Strongly expressed

24、EGF-IR appeared as a mosaic (M) pattern in the metastatic lymph node. ABC method 100Fig.7 EGFR-IR in normal human placenta tissues. ABC method 200Fig.8 Strongly expressed EGFR-IR appeared as a mosaic (M) pattern in human esophageal squamous cell carcinoma (grade ) with lymph node metastasis. ABC met

25、hod 400Fig.9 Faintly expressed EGFR-IR appeared as a diffuse (D) pattern in human esophageal squamous cell carcinoma (grade ) without lymph node metastasis. ABC method 200Fig.10 EGFR-IR appeared in some nucleus. ABC method 400Fig.11 EGFR-IR appeared as a mosaic partten in the metastatic lymph node.

26、ABC method 100Fig.12 EGFR-IR in normal human esophageal epithelia. ABC method 200 Department of Histology and Embryology, Henan Medical University, Zhengzhou 450052, China作者單位：丁明杰丁蔚白經(jīng)修張娓高曉東張林慶牛富文付士顯(河南醫(yī)科大學(xué)組織學(xué)胚胎學(xué)教研室，450052）銀德仁（新鄉(xiāng)市中心醫(yī)院病理科)參考文獻(xiàn)1Cohen S. Purification and metabolic effects of a nerve growth-promoting protein from snake venom. J Biol Chem, 1959,234(5):11292Ozawa S, Uede M, Ando N, et al. Epidermal growth factor receptors in cancer tissues of esophagus, lung, pancreas, colorectum, breast and stomach. Jpn J Cancer Res (Gann), 1988,79(11):12013陳元方