張繼君 石曉欣 陶麗麗 陳耀麗 趙昌林
[摘要]目的 探討叉頭盒A1(FOXA1)及AMACR/P504s在前列腺癌中的表達(dá)及其臨床意義。方法 收集2014年1月~2018年10月北京大學(xué)深圳醫(yī)院及深圳市第二人民醫(yī)院經(jīng)病理確診的49例原發(fā)性前列腺癌標(biāo)本及8例前列腺癌骨轉(zhuǎn)移標(biāo)本。采用免疫組化EnVision法檢測(cè)前列腺癌組織中FOXA1及AMACR/P504s的蛋白表達(dá),分析FOXA1和AMACR/P504s的表達(dá)情況。結(jié)果 FOXA1在前列腺癌轉(zhuǎn)移標(biāo)本中的陽(yáng)性率為62.5%(5/8),低于原發(fā)性前列腺癌的93.9%(46/49),差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。AMACR/P504s在前列腺癌轉(zhuǎn)移標(biāo)本中的陽(yáng)性率為75.0%(6/8),與原發(fā)性前列腺癌的83.7%(41/49)比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05)。原發(fā)性前列腺癌:T分期中,T3~4期的FOXA1陽(yáng)性表達(dá)率為67.3%,顯著高于T2a~c期的26.5%,差異有統(tǒng)計(jì)學(xué)意義(P<0.05);Gleason評(píng)分中,7~10分的FOXA1陽(yáng)性表達(dá)率為77.6%,顯著高于6分的16.3%,差異有統(tǒng)計(jì)學(xué)意義(P<0.05);有脈管浸潤(rùn)的FOXA1陽(yáng)性表達(dá)率為14.3%,顯著低于無(wú)脈管浸潤(rùn)的79.6%,差異有統(tǒng)計(jì)學(xué)意義(P<0.05);不同年齡、神經(jīng)束侵犯情況下的FOXA1陽(yáng)性表達(dá)率比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05);不同T分期、Gleason評(píng)分、年齡、脈管浸潤(rùn)以及神經(jīng)束侵犯情況下的AMACR/P504s陽(yáng)性表達(dá)率比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05)。結(jié)論 FOXA1和AMACR/P504s是前列腺癌良好的標(biāo)志物,F(xiàn)OXA1高表達(dá)提示預(yù)后差。
[關(guān)鍵詞]前列腺癌;叉頭盒;AMACR/P504s
[中圖分類號(hào)] R737.25 ? ? [文獻(xiàn)標(biāo)識(shí)碼] A ? ? [文章編號(hào)] 1674-4721(2019)9(b)-0007-04
Expression and clinical significance of FOXA1 and AMACR/P504s in prostate cancer
ZHANG Ji-jun1? ?SHI Xiao-xin2? ?TAO Li-li2? ?CHEN Yao-li2▲? ?ZHAO Chang-lin3
1. Department of Pathology, Nanshan District People′s Hospital of Shenzhen City, Guangdong Province, Shenzhen? ?518000, China; 2. Department of Pathology, Peking University Shenzhen Hospital, Guangdong Province, Shenzhen? ?518036, China; 3. Department of Oncology, Guangzhou University of Chinese Medicine Clifford Hospital, Guangdong Province, Guangzhou? ?511495, China
[Abstract] Objective To investigate the expression and clinical significance of FOXA1 and AMACR/P504s in prostate cancer. Methods A total of 49 specimens of primary prostate cancer and 8 specimens of bone metastases from prostate cancer confirmed by pathology were collected from Peking University Shenzhen Hospital and Shenzhen Second People′s Hospital from January 2014 to October 2018. Immunohistochemical EnVision method was used to detect the protein expression of FOXA1 and AMACR/P504s in prostate cancer tissues, and the expression of FOXA1 and AMACR/P504s was analyzed. Results The positive rate of FOXA1 in prostate cancer metastasis specimens was 62.5% (5/8), which was lower than that of primary prostate cancer accounting for 93.9% (46/49), and the difference was statistically significant (P<0.05). The positive rate of AMACR/P504s in prostate cancer metastasis specimens was 75.0% (6/8), compared with 83.7% (41/49) of primary prostate cancer, and the difference was not statistically significant (P>0.05). Primary prostate cancer: in the T stage, the positive expression rate of FOXA1 in T3-4 stage was 67.3%, which was significantly higher than that in T2a-c stage for 26.5%, and the difference was statistically significant (P<0.05). In the Gleason score, the positive expression rate of FOXA1 at 7-10 points was 77.6%, which was significantly higher than that of 6 points for 16.3%, and the difference was statistically significant (P<0.05). The positive expression rate of FOXA1 in vascular infiltration was 14.3%, which was significantly lower than that of no vessel infiltration (79.6%), and the difference was statistically significant (P<0.05). There was no significant difference in the positive expression rate of FOXA1 in different ages and nerve bundle intrusion (P>0.05). There was no significant difference in the positive expression rate of AMACR/P504s in different T stages, Gleason scores, ages, vascular infiltration and nerve bundle intrusion (P>0.05). Conclusion FOXA1 and AMACR/P504s are good markers for prostate cancer, and high expression of FOXA1 suggests poor prognosis.
[Key words] Prostate cancer; FOXA1;AMACR/P504s
前列腺癌是發(fā)達(dá)國(guó)家男性常見(jiàn)的惡性腫瘤[1-2],在我國(guó)的發(fā)病率也迅速升高,其腫瘤進(jìn)展的分子學(xué)機(jī)制尚不完全清楚。AMACR酶參與過(guò)氧化物酶體-氧化支鏈脂肪酸及其衍生物的代謝,已經(jīng)發(fā)現(xiàn)AMACR/P504s在前列腺癌組織內(nèi)其mRNA及蛋白均升高。叉頭盒A1(FOXA1)又被稱為肝細(xì)胞核因子3a(HNF3a),是FOX家族轉(zhuǎn)錄因子成員[3],能夠與非轉(zhuǎn)錄狀態(tài)的染色體結(jié)合,提高轉(zhuǎn)錄起始因子的基因表達(dá)及轉(zhuǎn)錄效率,從而調(diào)節(jié)下游基因的表達(dá)[4]。研究表明,F(xiàn)OXA1在多種癌癥中高表達(dá),包括涎腺導(dǎo)管癌[5]、非小細(xì)胞肺癌[6]、食管癌[7]、甲狀腺癌[8]及乳腺癌[9-10]。FOXA因子在器官包括前列腺[11-12]的發(fā)育中具有重要作用,可影響前列腺癌的增殖和激素應(yīng)答[13-14]。本研究通過(guò)免疫組化EnVision法檢測(cè)FOXA1與AMACR/P504s在原發(fā)性及轉(zhuǎn)移性前列腺癌組織中的表達(dá),分析其與臨床的關(guān)系,評(píng)估FOXA1與AMACR/P504s在前列腺癌預(yù)后的價(jià)值,現(xiàn)報(bào)道如下。
1資料與方法
1.1一般資料
收集2014年1月~2018年10月北京大學(xué)深圳醫(yī)院及深圳市第二人民醫(yī)院經(jīng)病理確診的49例原發(fā)性前列腺癌標(biāo)本及8例前列腺癌骨轉(zhuǎn)移標(biāo)本。49例原發(fā)性前列腺癌標(biāo)本取材前均無(wú)化療或放療病史。57例患者,年齡52~79歲,平均(67.4±14.4)歲;<64歲20例,≥64歲37例;病理類型:前列腺腺癌56例,前列腺小細(xì)胞癌1例。原發(fā)性前列腺癌中,T分期:T2a~c 14例,T3~4 35例;Gleason評(píng)分:6分11例,7分16例,8~10分22例;年齡:<64歲19例,≥64歲30例;脈管浸潤(rùn):有7例,無(wú)42例;神經(jīng)束侵犯:有28例,無(wú)21例。
1.2方法
標(biāo)本采用10%中性福爾馬林固定,經(jīng)脫水、石蠟包埋,4 μm厚度切片及免疫組化EnVision兩步法檢測(cè)。FOXA1和AMACR/P504s分別為鼠抗、兔抗人單克隆抗體,均購(gòu)于福州邁新公司。具體操作步驟按照試劑盒說(shuō)明書(shū)進(jìn)行,其中AMACR/P504s為即用型試劑,F(xiàn)OXA1為濃縮液抗體,稀釋濃度為1:100。PBS代替一抗作為空白對(duì)照,采用DAB顯色,蘇木精對(duì)比染色。
1.3結(jié)果判定標(biāo)準(zhǔn)
FOXA1蛋白在細(xì)胞核表達(dá)。AMACR/P504s陽(yáng)性染色為彌漫性或粒狀、胞漿性或腔內(nèi)染色。AMACR/P504s在相鄰的良性腺體如有染色,可見(jiàn)部分弱染色[15]。陰性染色屬于無(wú)染色或局灶性、弱的非腔緣細(xì)顆粒染色,對(duì)染色強(qiáng)度及陽(yáng)性細(xì)胞百分?jǐn)?shù)分別進(jìn)行評(píng)判。①按陽(yáng)性腫瘤細(xì)胞百分?jǐn)?shù)評(píng)分:陽(yáng)性細(xì)胞數(shù)占1%~10%為1分;11%~50%為2分;51~80%為3分;81%~100%為4分。②按染色強(qiáng)度評(píng)分:陰性為0分;弱陽(yáng)性為1分;中度陽(yáng)性為2分;強(qiáng)陽(yáng)性為3分。將兩項(xiàng)得分之積作為最終得分,總分為0~12分,其中0分為陰性,其余均為陽(yáng)性[16]。
1.4統(tǒng)計(jì)學(xué)方法
采用SPSS 19.0統(tǒng)計(jì)學(xué)軟件進(jìn)行數(shù)據(jù)分析,計(jì)量資料用均數(shù)±標(biāo)準(zhǔn)差(x±s)表示,兩組間比較采用t檢驗(yàn);計(jì)數(shù)資料采用率表示,組間比較采用χ2檢驗(yàn)或Fisher精確檢驗(yàn),以P<0.05為差異有統(tǒng)計(jì)學(xué)意義。
2結(jié)果
2.1 FOXA1和AMACR/P504s在原發(fā)性前列腺癌中的表達(dá)情況
FOXA1僅存在于上皮細(xì)胞細(xì)胞核,在正常和腫瘤組織間質(zhì)中均未觀察到免疫反應(yīng)。腫瘤旁前列腺周圍帶組織內(nèi)有弱至中等程度表達(dá),腫瘤旁前列腺移行帶組織內(nèi)FOXA1蛋白幾乎不表達(dá)(圖1A);AMACR/P504s在腫瘤旁正常前列腺組織不表達(dá)(圖2A),F(xiàn)OXA1與AMACR/P504s蛋白在前列腺癌及前列腺上皮內(nèi)病變均有表達(dá)(圖1B~C、圖2B~C)。FOXA1與AMACR/P504s在原發(fā)性前列腺癌中的陽(yáng)性率分別為93.9%(46/49)和83.7%(41/49),兩者的陽(yáng)性率比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05)。
2.2 FOXA1和AMACR/P504s在前列腺癌骨轉(zhuǎn)移組織中的表達(dá)情況
FOXA1在前列腺癌轉(zhuǎn)移標(biāo)本中的陽(yáng)性率為62.5%(5/8)(圖3A),低于原發(fā)性前列腺癌的93.9%(46/49),差異有統(tǒng)計(jì)學(xué)意義(P<0.05);AMACR/P504s在前列腺癌轉(zhuǎn)移標(biāo)本中的陽(yáng)性率為75.0%(6/8)(圖3B),與原發(fā)性前列腺癌的83.7%(41/49)比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05);FOXA1蛋白在前列腺小細(xì)胞癌中表達(dá)較強(qiáng)(圖3C)。
2.3 FOXA1和AMACR/P504s在原發(fā)性前列腺癌組織不同病理參數(shù)情況下的表達(dá)
T分期中,T3~4期的FOXA1陽(yáng)性表達(dá)率為67.3%,顯著高于T2a~c期的26.5%,差異有統(tǒng)計(jì)學(xué)意義(P<0.05);Gleason評(píng)分中,7~10分的FOXA1陽(yáng)性表達(dá)率為77.6%,顯著高于6分的16.3%,差異有統(tǒng)計(jì)學(xué)意義(P<0.05);有脈管浸潤(rùn)的FOXA1陽(yáng)性表達(dá)率為14.3%,顯著低于無(wú)脈管浸潤(rùn)的79.6%,差異有統(tǒng)計(jì)學(xué)意義(P<0.05);不同年齡、神經(jīng)束侵犯情況下的FOXA1陽(yáng)性表達(dá)率比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05);不同T分期、Gleason評(píng)分、年齡、脈管浸潤(rùn)以及神經(jīng)束侵犯情況下的AMACR/P504s陽(yáng)性表達(dá)率比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05)(表1)。
3討論
FOXA1和AMACR/P504s可以作為前列腺癌病理診斷的標(biāo)志物。FOXA1具有調(diào)節(jié)類固醇激素受體轉(zhuǎn)活化的功能,是前列腺上皮分化所必需的轉(zhuǎn)錄因子[17]。小鼠模型研究表明FOXA1在前列腺上皮內(nèi)瘤變和前列腺腺癌中的表達(dá)持續(xù)上調(diào)[18]。本研究結(jié)果提示,F(xiàn)OXA1在前列腺癌及前列腺上皮內(nèi)病變中表達(dá)均上調(diào),AMACR/P504s在前列腺癌及其上皮內(nèi)病變組織內(nèi)高表達(dá)[19-21],F(xiàn)OXA1與AMACR/P504s在原發(fā)性前列腺癌中的陽(yáng)性率比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05)。兩種蛋白在前列腺癌的診斷中各有優(yōu)缺點(diǎn),一方面AMACR/P504s的特異性高于FOXA1,在正常前列腺組織中幾乎不表達(dá),在前列腺癌中高表達(dá);而FOXA1除了在前列腺癌中高表達(dá),在正常前列腺周圍帶組織內(nèi)也有一定的表達(dá)。本研究共選擇57例前列腺癌組織(包括原發(fā)性及前列腺癌轉(zhuǎn)移標(biāo)本),聯(lián)合應(yīng)用FOXA1與AMACR/P504s兩種標(biāo)志物的診斷率達(dá)100.0%,尤其是聯(lián)合應(yīng)用這兩種抗體對(duì)鑒別診斷轉(zhuǎn)移性前列腺癌與其他來(lái)源腫瘤具有非常重要的臨床意義。FOXA1蛋白表達(dá)在腫瘤細(xì)胞胞核,而AMACR/P504s蛋白表達(dá)在腫瘤細(xì)胞胞漿,兩種蛋白的表達(dá)模式有利于應(yīng)用雞尾酒染色法同時(shí)標(biāo)記腫瘤細(xì)胞。兩者可作為前列腺癌診斷的標(biāo)志物。
FOXA1高表達(dá)提示疾病分期晚、惡性程度高、預(yù)后差。本研究結(jié)果顯示,不同T分期、Gleason評(píng)分、脈管浸潤(rùn)情況下的FOXA1陽(yáng)性表達(dá)率比較,差異有統(tǒng)計(jì)學(xué)意義(P<0.05);不同年齡、神經(jīng)束侵犯情況下的FOXA1陽(yáng)性表達(dá)率比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05);不同T分期、Gleason評(píng)分、年齡、脈管浸潤(rùn)以及神經(jīng)束侵犯情況下的AMACR/P504s陽(yáng)性表達(dá)率比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05)。提示Gleason評(píng)分高、T分期晚,F(xiàn)OXA1表達(dá)越強(qiáng),F(xiàn)OXA1有可能成為一種新的治療靶點(diǎn)。
綜上所述,F(xiàn)OXA1和AMACR/P504s是前列腺癌良好的標(biāo)志物。FOXA1高表達(dá)提示預(yù)后差。本研究缺乏去勢(shì)抵抗性前列腺癌(CRPC)標(biāo)本,尚不清楚FOXA1與AMACR/P504s在這一類腫瘤中表達(dá)情況,有待進(jìn)一步的研究。
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(收稿日期:2019-03-06? 本文編輯:任秀蘭)