張?zhí)毂耄尬妮x，趙瀅，陳鑫瑩

1.中國醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院生物化學(xué)與分子生物學(xué)教研室，遼寧 沈陽 110122；

2.中國醫(yī)科大學(xué)附屬盛京醫(yī)院胃腸營養(yǎng)外科，遼寧 沈陽，110004

S100鈣結(jié)合蛋白A4在胃癌組織中的表達(dá)及意義

張?zhí)毂?，宿文輝1，趙瀅2，陳鑫瑩2

1.中國醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院生物化學(xué)與分子生物學(xué)教研室，遼寧 沈陽 110122；

2.中國醫(yī)科大學(xué)附屬盛京醫(yī)院胃腸營養(yǎng)外科，遼寧 沈陽，110004

背景與目的：本研究探討S100鈣結(jié)合蛋白A4(S100 calcium-binding protein A4，S100A4)在胃癌旁正常組織、慢性胃炎、腸上皮化生、腺瘤樣異型增生和胃癌的組織標(biāo)本中的表達(dá)及與胃癌臨床病理特征及預(yù)后的關(guān)系。方法：利用HE染色對(duì)所取胃組織標(biāo)本進(jìn)行病理組織學(xué)診斷；采用免疫組織化學(xué)方法檢測(cè)標(biāo)本S100A4蛋白的表達(dá)；實(shí)時(shí)定量聚合酶鏈反應(yīng)(quantitative real-time pdymerase chain reaction，qRT-PCR)法檢測(cè)S100A4 mRNA表達(dá)；蛋白[質(zhì)]印跡法(Western blot)檢測(cè)S100A4蛋白的表達(dá)；采用Kaplan-Meier分析累積生存率。結(jié)果：S100A4蛋白和mRNA的表達(dá)按以下順序逐漸增加：胃癌旁正常組織＜慢性胃炎＜腸上皮化生＜腺瘤樣異型增生＜胃癌。S100A4表達(dá)水平與腫瘤大小、浸潤深度、淋巴管浸潤、淋巴結(jié)轉(zhuǎn)移和腫瘤TNM分期有關(guān)，但與患者的年齡和性別無關(guān)。在較大直徑、較深浸潤、淋巴結(jié)轉(zhuǎn)移的胃癌和腸型癌中可發(fā)現(xiàn)S100A4基因的過表達(dá)。采用Kaplan-Meier單因素分析顯示，弱或中度S100A4基因表達(dá)的患者與不表達(dá)的患者相比有較低的累積生存率。結(jié)論：在胃癌旁正常組織-慢性胃炎-腸上皮化生-腺瘤樣異型增生-胃癌過程中，S100A4 mRNA和蛋白表達(dá)逐漸增高；S100A4表達(dá)與胃癌腫瘤大小、侵襲深度、淋巴結(jié)轉(zhuǎn)移和不良預(yù)后呈正相關(guān)；S100A4過表達(dá)參與胃癌發(fā)生和演進(jìn)過程，可以作為反映胃癌惡性程度的生物學(xué)指標(biāo)。

S100鈣結(jié)合蛋白A4；胃癌；免疫組化；RT-PCR；蛋白[質(zhì)]跡法

胃癌是我國癌癥相關(guān)死亡率的第二大原因［1］，僅次于肺癌。自從20世紀(jì)下半葉以來，雖然胃癌的發(fā)病率及死亡率在世界范圍內(nèi)有下降趨勢(shì)，它仍是全球主要的健康問題［2］。因此，闡明胃癌的發(fā)生、發(fā)展的分子機(jī)制和可靠的生物標(biāo)記物的篩選對(duì)胃癌的預(yù)防、治療和預(yù)后評(píng)估至關(guān)重要的。

S100蛋白是鈣結(jié)合蛋白，呈酸性，相對(duì)分子質(zhì)量為10×103～12×103，由于其在100%飽和硫酸銨中完全溶解而命名。S100鈣結(jié)合蛋白A4 (S100 calcium-binding protein A4，S100A4)，又稱MTS1(轉(zhuǎn)移相關(guān)基因)等，其過表達(dá)，已被證明可以控制細(xì)胞周期進(jìn)程和調(diào)節(jié)細(xì)胞間黏附，以及腫瘤細(xì)胞的侵襲性和轉(zhuǎn)移性［3］。S100A4基因啟動(dòng)子含有一個(gè)ErbB2的應(yīng)答元件，而S100A4基因的增強(qiáng)子和沉默子元件可被甲基化強(qiáng)烈影響［4-5］。有報(bào)道稱S100A4在體外結(jié)合到p53的C-末端調(diào)節(jié)結(jié)構(gòu)域并抑制其磷酸化，從而抑制了p53的靶基因的轉(zhuǎn)錄，包括p21/WAF、Bax蛋白、血小板反應(yīng)蛋白-1和MDM-2［6］。S100A4還參與細(xì)胞骨架重排和細(xì)胞運(yùn)動(dòng)，如F-肌動(dòng)蛋白、肌球蛋白-ⅡA、原肌球蛋白和非肌肉肌球蛋白Ⅱ的重鏈［7-9］。此外，S100A4已經(jīng)表明通過其對(duì)血管形成、細(xì)胞骨架完整性、基質(zhì)金屬蛋白酶(MMPs)的活性、腫瘤相關(guān)的轉(zhuǎn)錄因子和基質(zhì)因子的影響，以呈現(xiàn)其轉(zhuǎn)移前作用［10-11］。

本研究的目的是分析胃癌及癌前病變中S100A4基因在蛋白和mRNA水平的表達(dá)，并進(jìn)一步比較S100A4基因的表達(dá)與胃癌的臨床病理特征及預(yù)后的關(guān)系。

1 材料和方法

1.1 材料

收集中國醫(yī)科大學(xué)附屬盛京醫(yī)院2005年1月—2015年1月胃組織標(biāo)本，內(nèi)鏡活檢組織中收集慢性胃炎(73例)，胃黏膜腸上皮化生(86例)和腺瘤樣異型增生(63例)，胃癌及癌旁正常組織標(biāo)本(各348例)來源于胃手術(shù)切除組織。所有患者在手術(shù)前未接受化療，放療或輔助治療。蘇木精和伊紅染色(hematoxylin and eosin，HE)確認(rèn)病理組織學(xué)診斷。標(biāo)本的部分組織凍結(jié)在-80 ℃液氮中用于隨后RNA和蛋白質(zhì)的提取。根據(jù)國際抗癌聯(lián)盟(Union Internationale Contre le Cancer，UICC)標(biāo)準(zhǔn)對(duì)胃癌標(biāo)本進(jìn)行TNM(tumornode-metastasis)分期［12］，病理組織學(xué)分型依照Lauren’s分型［13-14］的判定，胃癌細(xì)胞淋巴管和靜脈侵犯由HE染色評(píng)估，必要時(shí)進(jìn)行D2-40免疫染色和EVG染色。預(yù)后統(tǒng)計(jì)通過門診復(fù)查和電話隨訪方式進(jìn)行。

1.2 方法

1.2.1 組織芯片制備及免疫組化分析

收集2005年1月—2015年1月存檔蠟塊作為所需的供體蠟塊，組織標(biāo)本在4%的中性甲醛溶液固定，石蠟包埋，切成4 μm的石蠟切片。顯微鏡下觀察相應(yīng)HE切片確定具有代表性的病變部位。選取時(shí)應(yīng)避開壞死較多處。將優(yōu)質(zhì)石蠟和蜂蠟按100∶l混合，經(jīng)多次沉淀去除雜質(zhì)鑄成具有較好柔韌度和硬度的蠟塊。利用Tissue Arrayer對(duì)受體蠟塊進(jìn)行打孔(直徑2 mm)，而后在選取的供體蠟塊中穿取標(biāo)記組織(直徑2 mm)，準(zhǔn)確放入受體蠟塊的小孔中。按序依次操作，同時(shí)作記錄。將受體蠟塊放入50 ℃溫箱中烤20～40 min，待石蠟軟化，使供體蠟塊組織和受體蠟塊均勻融合。石蠟切片機(jī)連續(xù)切片，厚度4 μm，敷貼于10%多聚賴氨酸預(yù)先處理的載玻片上，低溫保存。采用EnVision方法對(duì)胃癌及其癌前病變組織芯片進(jìn)行免疫組化染色。兔抗人S100A4多克隆抗體(工作濃度1∶200)購自Abcom公司。所有步驟依據(jù)產(chǎn)品說明進(jìn)行，并利用PBS(0.01 mol/L，pH值為7.4)替代一抗作為陰性對(duì)照。

免疫組化染色結(jié)果判定：免疫組化切片隨機(jī)分配給兩名有經(jīng)驗(yàn)的病理學(xué)醫(yī)生獨(dú)立分析，這兩名病理學(xué)醫(yī)生事先不了解患者的臨床資料。在200的放大倍率，來確定S100A4陽性細(xì)胞數(shù)，根據(jù)陽性染色細(xì)胞的百分比進(jìn)行半定量分級(jí)并根據(jù)評(píng)分系統(tǒng)分為四級(jí)：陰性(-)，0%～5%；弱陽性(+)，6%～25%；陽性(++)，26%～50%和強(qiáng)陽性(+++)＞50%。

1.2.2 蛋白［質(zhì)］印跡法(Western blot)檢測(cè)S100A4蛋白

組織蛋白提取后，采用BCA法進(jìn)行裂解液中蛋白濃度的測(cè)定，然后進(jìn)行常規(guī)的Western blot法操作。應(yīng)用凝膠成像分析軟件(Quantity One)進(jìn)行灰度掃描，通過目的蛋白與內(nèi)參照蛋白GAPDH的積分吸光度比值表示蛋白相對(duì)表達(dá)量。

1.2.3 實(shí)時(shí)定量聚合酶鏈反應(yīng)(quantitative realtime polymerase chain reaction，qRT-PCR)法檢測(cè)S100A4 mRNA

經(jīng)提取總R N A后進(jìn)行實(shí)時(shí)定量P C R反應(yīng)，引物序列S 1 0 0 A 4基因順義鏈為5'-AGCTTCTTGGGGAAAAGGAC-3’，反義鏈為5'-TACACATCATGGCGATGCAG-3'(引物長度均為130 bp)；GAPDH基因順義鏈為5'-CAATGACCCCTTCATTGACC-3'，反義鏈為5'-TGGAA GATGGTGATGGGATT-3'(引物長度均為135 bp)。

采用相對(duì)定量的方法，以GAPDH基因mRNA的表達(dá)為內(nèi)參基因，依據(jù)公式△Ct=Ct(目的基因)-Ct(內(nèi)參基因)，分別計(jì)算實(shí)驗(yàn)組和對(duì)照組的△Ct，再依公式△△Ct=△Ct(實(shí)驗(yàn)組)-△Ct(對(duì)照組)，計(jì)算出△△Ct值，最后計(jì)算相對(duì)表達(dá)量的差值即2-△△Ct，其表示的是實(shí)驗(yàn)組目的基因的表達(dá)相對(duì)于對(duì)照組的變化倍數(shù)。

1.3 統(tǒng)計(jì)學(xué)處理

統(tǒng)計(jì)分析采用χ2檢驗(yàn)或Fisher’s Exact Test比較計(jì)數(shù)資料的組間差異，Mann-Whitney Test檢驗(yàn)比較計(jì)量資料組間差異，Kaplan-Meier生存曲線用于區(qū)分和比較生存率。Cox比例風(fēng)險(xiǎn)模型用于多因素分析。應(yīng)用SPSS 10.0軟件(SPSS Inc., Chicago，IL，USA)分析數(shù)據(jù)，P＜0.05為差異有統(tǒng)計(jì)學(xué)意義。

2 結(jié) 果

2.1 免疫組化分析S100A4的表達(dá)

圖1 胃組織中的S100A4蛋白免疫組化染色(EnVision, ×100)Fig .1 Immunohistochemical staining of S100A4 protein showed positive expression in different gastric lesions

S100A4蛋白的表達(dá)通過測(cè)定腫瘤細(xì)胞陽性染色的數(shù)量采用四點(diǎn)計(jì)分制進(jìn)行評(píng)估。如圖1所示，S100A4蛋白定位于胃上皮細(xì)胞，腸上皮化生，腺瘤樣異型增生和胃癌細(xì)胞的細(xì)胞質(zhì)和細(xì)胞核中，出現(xiàn)棕黃色顆粒。S100A4蛋白陽性表達(dá)率為癌旁正常組織8.3%(29/348)，慢性胃炎19.2%(14/73)，腸上皮化生23.3%(20/86)，腺瘤樣異型增生34.9%(22/63)，胃癌55.2% (192/348)?；赟100A4表達(dá)的頻率和密度，我們推斷S100A4蛋白的表達(dá)按以下順序逐漸增加：癌旁正常組織＜慢性胃炎＜腸上皮化生＜腺瘤樣異型增生＜胃癌(P＜0.001，表1)。

S100A4蛋白的表達(dá)與臨床病理特征之間的相關(guān)性分析，分析S100A4蛋白陽性表達(dá)與臨床病理特征之間的相關(guān)性及可能影響胃癌患者預(yù)后的因素。結(jié)果發(fā)現(xiàn)，S100A4基因的表達(dá)與腫瘤大小、浸潤深度、淋巴管和靜脈侵犯、淋巴結(jié)轉(zhuǎn)移，腫瘤TNM分期(P＜0.05)呈正相關(guān)，但是患者的年齡和性別對(duì)S100A4表達(dá)無影響(P＜0.05)。S100A4基因表達(dá)在腸型(IT)胃癌高于彌漫型(DT)癌(P＜0.001，表2)。

2.2 qRT-PCR和Western blot分析S100A4表達(dá)與胃癌臨床病理特征相關(guān)性的結(jié)果

S100A4 mRNA的表達(dá)按以下順序上調(diào)：慢性胃炎＜腸上皮化生＜腺瘤樣異型增生＜胃癌(P＜0.001，圖2，表3)。本研究發(fā)現(xiàn)在直徑較大(＞4 cm)，浸潤較深及較多淋巴結(jié)轉(zhuǎn)移的胃癌組織中S100A4 mRNA呈現(xiàn)高表達(dá)(P＜0.05)，也支持Western blot分析S100A4蛋白的表達(dá)結(jié)果(圖3，表4)。

表1 胃組織中S100A4蛋白的表達(dá)Tab. 1 Differential expression of S100A4 protein in gastric tissue specimens

表2 胃癌組織中S100A4蛋白的表達(dá)與臨床病理特征的關(guān)系Tab. 2 Association of S100A4 protein expression with the clinicopathological features of the gastric cancer patients

圖2 S100A4 mRNA的表達(dá)水平與胃癌臨床病理特征的關(guān)系Fig. 2 Association of S100A4 mRNA expression with the clinicopathological features of the gastric cancer patients

表3 實(shí)時(shí)RT-PCR檢測(cè)胃癌組織中S100A4蛋白的表達(dá)與臨床病理特征的關(guān)系Tab. 3 Association of S100A4 protein expression with the clinicopathological features of the gastric cancer patients by RT-PCR

表4 Western blot檢測(cè)胃癌組織中S100A4蛋白的表達(dá)與臨床病理特征的關(guān)系Tab. 4 Association of S100A4 protein expression with the clinicopathological features of the gastric cancer patients by Western blot

圖3 S100A4蛋白的表達(dá)水平與胃癌臨床病理特征的關(guān)系Fig. 3 S100A4 protein expression level during gastric carcinogenesis and its correlation with clinicopathological features of carcinoma.

2.3 S100A4蛋白表達(dá)與胃癌患者生存的關(guān)系

對(duì)348例胃癌患者進(jìn)行為期 1個(gè)月～10.1年(中位數(shù)65.1個(gè)月)的隨訪。Kaplan-Meier法單因素分析顯示：弱或中度S100A4基因表達(dá)的患者累積生存率明顯低于S100A4基因不表達(dá)的患者(P＜0.05)。多因素分析采用Cox比例模型來確定各種臨床病理參數(shù)的獨(dú)立預(yù)后作用。研究結(jié)果表明，浸潤深度(P=0.048)，淋巴管及靜脈浸潤(P=0.049和0.041)，淋巴結(jié)轉(zhuǎn)移(P=0.027)，TNM分期(P＜0.001)與S100A4表達(dá)(P=0.045)均是胃癌的預(yù)后不良的獨(dú)立因素(表5，圖4)。

圖4 S100A4蛋白的表達(dá)與胃癌患者預(yù)后的關(guān)系Fig. 4 Correlation between S100A4 expression status and prognosis of gastric carcinoma patients

表5 胃癌患者生存的多因素分析Tab. 5 Multivariate analysis of clinicopathological variables to determine the survival of gastric carcinoma cases.

3 討 論

S100A4蛋白定位于胃上皮細(xì)胞的細(xì)胞質(zhì)和細(xì)胞核，這與以前的報(bào)道一致［15-16］。 S100A4是一個(gè)轉(zhuǎn)錄因子，可以在某些生理或病理?xiàng)l件下從細(xì)胞核轉(zhuǎn)運(yùn)到細(xì)胞質(zhì)。在本研究中S100A4 mRNA和蛋白的表達(dá)水平按以下順序逐漸增加：癌旁組織＜慢性胃炎＜腸上皮化生＜腺瘤樣異型增生＜胃癌，與腎透明細(xì)胞癌［17］，食管鱗狀細(xì)胞癌［18-19］，大腸癌［20］，乳腺癌［21］和膀胱癌［22］一致。腸上皮化生是一種損傷的胃上皮的自然適應(yīng)狀態(tài)，根據(jù)其外觀形態(tài)和生物學(xué)特性，可發(fā)展成球樣異型增生，與印戒細(xì)胞癌的發(fā)生密切相關(guān)［23］。病理和遺傳學(xué)觀察表明，胃黏膜異型增生的發(fā)生早于胃癌，并分為隱窩、球樣再生和腺瘤亞型［24］。有報(bào)道稱［25-26］，S100A4超表達(dá)可能導(dǎo)致S100A4基因第一個(gè)內(nèi)含子的CpG位點(diǎn)發(fā)生甲基化，或者只是在一個(gè)缺氧的微環(huán)境中S100A4刺激的結(jié)果。

通過分析S100A4的表達(dá)是否與胃癌的侵襲行為有關(guān)來闡明S100A4蛋白在胃癌進(jìn)展中的作用，本研究中S100A4 mRNA和蛋白表達(dá)與腫瘤大小、浸潤深度、淋巴管和靜脈侵襲、淋巴結(jié)轉(zhuǎn)移和TNM分期呈正相關(guān)，這支持Feng等［27］以前的結(jié)果。細(xì)胞質(zhì)中S100A4蛋白的表達(dá)與胃癌浸潤深度和淋巴結(jié)轉(zhuǎn)移呈正相關(guān)，S100A4蛋白的核表達(dá)與結(jié)直腸癌浸潤深度和嗜神經(jīng)侵襲呈正相關(guān)［28］。晚期子宮內(nèi)膜癌［29］和膀胱癌［30］中S100A4基因的表達(dá)和侵襲性之間呈正相關(guān)。這些結(jié)果表明，S100A4基因的超表達(dá)參與了胃癌的生長、侵襲和轉(zhuǎn)移，可作為臨床實(shí)踐中胃癌的侵襲性的指標(biāo)。因此，推測(cè)S100A4基因表達(dá)上調(diào)可能通過逆轉(zhuǎn)癌細(xì)胞的侵犯性表型參與胃癌的進(jìn)展。

S100A4基因是通過下調(diào)E-cadherin表達(dá)與調(diào)控AKT/Slug通路［19,30］，以調(diào)節(jié)人體食管鱗癌細(xì)胞的遷移和侵襲性行為。Sack等［31］報(bào)道，S100A4誘導(dǎo)的細(xì)胞運(yùn)動(dòng)和轉(zhuǎn)移是通過在結(jié)腸癌細(xì)胞中的Wnt/β-catenin蛋白通路抑制劑卡西霉素所抑制的。Zhang等［18］報(bào)道，S100A4蛋白介導(dǎo)的細(xì)胞侵襲和食管鱗狀細(xì)胞癌轉(zhuǎn)移是通過MMP-2和E-鈣粘蛋白的活性的調(diào)節(jié)。Shen等［32］發(fā)現(xiàn)，S100A4通過αV和α5整合調(diào)節(jié)防止胃癌細(xì)胞的凋亡。Hua等［33］發(fā)現(xiàn)，抑制S100A4蛋白促進(jìn)細(xì)胞凋亡并抑制BGC823胃癌細(xì)胞在體外和體內(nèi)的增殖。Xie等［34］發(fā)現(xiàn)，S100A4介導(dǎo)的子宮內(nèi)膜癌侵襲和誘導(dǎo)S100A4與腫瘤生長因子(TGF)-β1信號(hào)傳導(dǎo)的Smads蛋白激活相關(guān)?；谶@些數(shù)據(jù)和我們目前的研究結(jié)果，推測(cè)S100A4通過Wnt信號(hào)/β-catenin蛋白或TGF-β信號(hào)通路上調(diào)，增強(qiáng)了侵犯性的表型和惡性腫瘤的行為。

雖然胃癌起源于相同的胃上皮細(xì)胞，但由于患者的個(gè)體差異，其形態(tài)特征大不相同。根據(jù)Lauren分型，腸型胃癌以凝聚的腫瘤細(xì)胞所構(gòu)成的腺樣管狀結(jié)構(gòu)為特征，呈膨脹或浸潤性生長，包括高、中分化腺癌［7］。相反，彌漫型胃癌則以不明顯的腺癌或缺乏細(xì)胞黏附為主要特征，包括低分化和印戒細(xì)胞癌［23］。在本研究中，S100A4的蛋白和mRNA在腸型中表達(dá)明顯低于彌漫型，表明S100A4可能在其中發(fā)揮了重要致癌作用，可作為這兩種類型胃癌之間區(qū)別的分子基礎(chǔ)。

在本研究中，S100A4基因的表達(dá)與胃癌患者預(yù)后良好呈負(fù)相關(guān)，這與以前的報(bào)道相一致［15-16］。Cox比例風(fēng)險(xiǎn)分析表明，浸潤深度，淋巴管或靜脈侵襲，淋巴結(jié)轉(zhuǎn)移，TNM分期和S100A4表達(dá)是胃癌患者預(yù)后不良的獨(dú)立因素。Kho等［35］發(fā)現(xiàn)，C期結(jié)腸癌患者只有細(xì)胞質(zhì)出現(xiàn)S100A4的染色，細(xì)胞核不染色，S100A表達(dá)與患者的預(yù)后不良相關(guān)。Kang等［28］認(rèn)為大腸癌患者細(xì)胞核S100A4蛋白的表達(dá)是預(yù)后不良的獨(dú)立因素。類似的結(jié)果也出現(xiàn)在胰腺癌［36］、腎細(xì)胞癌［37］、膀胱癌［22］等。這些結(jié)果表明，S100A4基因表達(dá)是胃癌患者預(yù)后不良的獨(dú)立和可靠的指標(biāo)。

綜上所述，S100A4基因的上調(diào)可能在胃上皮細(xì)胞的惡性轉(zhuǎn)化中起重要作用，與胃癌的生長、侵襲、轉(zhuǎn)移和不良預(yù)后呈正相關(guān)。S100A4可被視為胃癌侵犯行為的一個(gè)有力的標(biāo)志物。S100A4在彌漫型癌的獨(dú)特表達(dá)可以用來區(qū)分腸型癌和彌漫型癌。

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S100 calcium binding protein A4 expression and significance in gastric cancer

ZHANG Tianbiao1, SU Wenhui1, ZHAO Ying2, CHEN Xinying2 (1. Department of Biochemistry and Molecular Biology, China Medical University, Shenyang Liaoning 110122, China; 2. Department of General Surgery, Shengjing Hospital of China Medical University, Shenyang Liaoning 110004, China)

ZHANG Tianbiao E-mail: tbzhang@mail.cmu.edu.cn

Background and purpose:This study investigated the relationship between (S100 calcium-binding protein A4, S100A4) in chronic gastritis, intestinal metaplasia, dysplasia adenomatous, normal tissue tissue samples and expression in gastric cancer and clinical characteristics.Methods:HE staining of the use of gastric specimens taken for histopathological diagnosis; using immunohistochemistry to detect the expression of tissue S100A4 protein; qRT-PCR was used to detect mRNA expression of S100A4 gene; Western Blot detection of S100A4 gene encoding protein. Kaplan-Meier survival curves were used to distinguish and compare survival.Results:S100A4 protein and mRNA expression gradually increased in the following order: normal tissue＜gastritis＜intestinal metaplasia＜dysplasia＜carcinoma. S100A4 levels with tumor size, depth of invasion, lymphatic invasion, lymph node metastasis and lymph node metastasis (TNM) staging, but with cancer patients independent of age and gender. Intestinal type (IT) cancer showed higher than the diffuse type (DT) of cancer S100A4 expression. S100A4 gene overexpression in a larger diameter, deeper invasion, lymph node metastasis of gastric and IT cancer was found. Kaplan-Meier method using univariate analysis showed weak expression of S100A4 gene is not expressed in patients with moderate or S100A4 gene have lower survival rates compared.Conclusion:Upregulated S100A4 gene may play an important role in the malignant transformation of epithelial cells in the stomach, and gastric cancer growth, invasion, metastasis and prognosis were positively correlated.Thus, S100A4 may be considered as a potential marker to show violations of gastric cancer. S100A4 expression in diffuse unique cancer can be used to distinguish between cancer and diffuse intestinal cancer, and can be used as a molecular characteristic distinguish two types of cancer.

S100 calcium-binding protein A4; Gastric cancer; Immunohistochemistry; RT-PCR; Western blot

10.3969/j.issn.1007-3969.2015.06.004

R735.2

A

1007-3639(2015)06-0424-09

2015-01-04

2015-04-30）

遼寧省科技廳基金(2013225021)。

張?zhí)毂?E-mail:tbzhang@mail.cmu.edu.cn