胃癌病人血清與癌組織nm23H1蛋白表達(dá)及臨床病理意義

宋培鐸,洪光晨

(青島市市南區(qū)人民醫(yī)院普外科,山東青島 266002)

胃癌病人血清與癌組織nm23H1蛋白表達(dá)及臨床病理意義

宋培鐸,洪光晨

(青島市市南區(qū)人民醫(yī)院普外科,山東青島 266002)

目的研究胃癌病人血清、癌組織中nm23H1蛋白的表達(dá)及與臨床病理特征的關(guān)系。方法采用ELISA法及免疫組織化學(xué)S-P法,檢測(cè)69例胃癌病人血清、癌組織中nm23 H1蛋白的表達(dá)情況,并分析其與臨床病理特征的關(guān)系。結(jié)果胃癌病人血清、癌組織中nm23H1蛋白的表達(dá)與正常對(duì)照組比較差異有顯著性(χ2＝9.69,t＝6.96,P＜0.05)。胃癌病人血清、癌組織中nm23H1蛋白的表達(dá)與淋巴結(jié)轉(zhuǎn)移及腫瘤臨床分期有關(guān)(χ2＝12.30、5.30,t＝14.67、6.42,P＜0.05)。結(jié)論胃癌病人血清及癌組織中nm23H1蛋白的表達(dá)下調(diào),nm23H1蛋白可以作為評(píng)估胃癌病人預(yù)后的指標(biāo)。

胃腫瘤;nm23H1;預(yù)后

腫瘤的侵襲、轉(zhuǎn)移是目前研究的一個(gè)熱點(diǎn)問題[1-6],發(fā)病率和死亡率均占我國(guó)消化道腫瘤首位的胃癌,其生物學(xué)行為具侵襲性,總體療效近幾年并未獲明顯改善。腫瘤的生長(zhǎng)分化和侵襲轉(zhuǎn)移是影響胃癌預(yù)后的主要因素,有多種基因參與調(diào)控胃癌的各項(xiàng)復(fù)雜生物學(xué)行為。除TNM分期外,目前能準(zhǔn)確判斷胃癌預(yù)后的指標(biāo)較少。由于影像學(xué)檢查等手段難以準(zhǔn)確判斷淋巴結(jié)的轉(zhuǎn)移情況,術(shù)前無法對(duì)胃癌做出準(zhǔn)確的TNM分期。本研究旨在探討胃癌病人血清、癌組織中nm23 H1蛋白的表達(dá)及其與臨床病理特征的關(guān)系?，F(xiàn)將結(jié)果報(bào)告如下。

1 材料與方法

1.1 標(biāo)本來源

2009年5月—2010年10月,我院手術(shù)切除的胃癌病人的標(biāo)本69例,男49例,女20例;平均年齡65.1歲。入選標(biāo)準(zhǔn):術(shù)前病人均未行化、放療;有完整的臨床、病理及隨訪資料。Lauren分型:腸型31例,彌漫型28例。伴淋巴結(jié)轉(zhuǎn)移者51例。臨床病理分期采用UICC胃癌國(guó)際統(tǒng)一TNM新分期法:Ⅰa期6例,Ⅰb期13例,Ⅱ期16例,Ⅲa期17例,Ⅲb期12例,Ⅳ期5例。并隨機(jī)選取同期就診的非腫瘤病人20例作為正常對(duì)照組。

1.2 實(shí)驗(yàn)方法

分別采集受檢者治療前不抗凝靜脈血2 m L,置室溫自然凝固后,離心10 min,取上清液置于低溫冰箱中備用。以全自動(dòng)生化分析儀,采用雙抗體夾心酶聯(lián)免疫吸附試驗(yàn)法(ELISA)檢測(cè)nm23H1蛋白的水平。實(shí)驗(yàn)操作及結(jié)果判定均嚴(yán)格按照試劑盒說明書的要求并由專人完成。采用免疫組化S-P法檢測(cè)腫瘤組織中nm23 H1蛋白的表達(dá)。石蠟切片常規(guī)脫蠟至水,室溫孵育30 min,微波修復(fù)抗原,以正常羊血清室溫孵育10 min,傾去血清,滴加一抗工作液,PBS沖洗。37℃溫箱孵育120 min,滴加二抗工作液,37℃溫箱孵育30 min,滴加鏈霉菌抗生物素-過氧化物酶溶液,37℃孵育10 min,DAB顯色,封固,顯微鏡觀察。每次實(shí)驗(yàn)用已知陽(yáng)性切片作為陽(yáng)性對(duì)照,以PBS代替一抗作為陰性對(duì)照。

1.3 結(jié)果判定

隨機(jī)選擇20個(gè)視野,高倍鏡下計(jì)數(shù)500～1 000個(gè)細(xì)胞中的陽(yáng)性細(xì)胞,計(jì)算nm23H1陽(yáng)性細(xì)胞百分率。根據(jù)陽(yáng)性細(xì)胞數(shù)量及顯色強(qiáng)弱分為:陰性,陽(yáng)性細(xì)胞＜10%;陽(yáng)性,陽(yáng)性細(xì)胞≥10%。

1.4 統(tǒng)計(jì)學(xué)分析

應(yīng)用SPSS 12.0軟件進(jìn)行分析。計(jì)數(shù)資料組間比較采用χ2檢驗(yàn),計(jì)量資料組間比較采用t檢驗(yàn)。

2 結(jié) 果

2.1 兩組nm23H1蛋白的表達(dá)

胃癌組織中nm23H1蛋白表達(dá)陽(yáng)性與陰性分別為32、37例,正常對(duì)照組分別為18、2例;胃癌組血清中nm23H1蛋白水平為(361.14±129.79)μg/ L,正常對(duì)照組為(570.87±64.96)μg/L,胃癌病人血清、癌組織中nm23 H1蛋白的表達(dá)與正常對(duì)照組比較差異有顯著性(χ2＝9.69,t＝6.96,P＜0.05)。

2.2 nm23 H1的表達(dá)與胃癌病人臨床特征的關(guān)系

nm23H1蛋白的表達(dá)與胃癌病人的年齡、性別、Lauren分型無關(guān)(P＞0.05),與病人淋巴結(jié)轉(zhuǎn)移及臨床TNM分期有關(guān)(χ2＝12.30、5.30,t＝14.67、6.42,P＜0.05)。見表1。

表1 胃癌病人血清及胃癌組織中nm23H1的表達(dá)與臨床病理特征的關(guān)系

3 討 論

作為一種轉(zhuǎn)移抑制基因,nm23倍受關(guān)注。許多研究結(jié)果表明,nm23基因的表達(dá)水平與乳癌、結(jié)腸直腸癌、肝癌、肺癌、黑色素瘤、喉鱗狀細(xì)胞癌、鼻咽癌、甲狀腺癌等腫瘤侵襲轉(zhuǎn)移呈顯著負(fù)相關(guān)[7-15]。nm23H1基因是1988年美國(guó)STEEG等[16]首次從小鼠K-1735黑色素瘤不同轉(zhuǎn)移潛能的細(xì)胞系中分離出來的。nm23 H1基因定位于染色體17q22[17], nm23H1蛋白的改變與腫瘤的浸潤(rùn)轉(zhuǎn)移有關(guān)[18]。KODERA等[19]對(duì)胃癌的研究結(jié)果顯示,nm23H1蛋白表達(dá)與淋巴結(jié)轉(zhuǎn)移呈負(fù)相關(guān)。胡建昆等[20]采用胃癌國(guó)際統(tǒng)一的新TNM分期法對(duì)343例胃癌病人進(jìn)行分析,結(jié)果Ⅰa、Ⅰb、Ⅱ、Ⅲa、Ⅲb及Ⅳ期的5年生存率分別為100%、82.69%、70.37%、48.44%、25.00%及7.69%,差異均有顯著性,分期與預(yù)后具有直接的關(guān)系。

本研究結(jié)果顯示,nm23H1蛋白的表達(dá)與胃癌病人的年齡、性別、Lauren分型無關(guān)。胃癌病人血清、癌組織中nm23H1蛋白的表達(dá)與正常對(duì)照組比較差異有顯著性,胃癌病人血清、癌組織中nm23 H1蛋白的表達(dá)與淋巴結(jié)轉(zhuǎn)移及腫瘤臨床分期有關(guān)。胃癌臨床TNM分期越晚,組織中nm23H1蛋白表達(dá)水平越低;無淋巴結(jié)轉(zhuǎn)移者nm23H1陽(yáng)性表達(dá)率明顯增高,而有淋巴結(jié)轉(zhuǎn)移者nm23H1陽(yáng)性表達(dá)率明顯降低,提示nm23H1蛋白表達(dá)水平越低預(yù)后可能越差。胃癌病人血清及癌組織中nm23 H1蛋白的表達(dá),可以作為評(píng)估胃癌病人預(yù)后的指標(biāo)。

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(本文編輯 厲建強(qiáng))

THE EXPRESSION AND ITS CLINICOPATHOLOGIC SIGNIFICANCE OF NM23H1 IN SERUM AND CANCER TISSUE OF PA-TIENTS WITH GASTRIC CANCER

SONG Peiduo,HONG Guangchen (The People’s Hospital of Qingdao Shinan District, Qingdao 266002,China)

ObjectiveTo study the expressions of nm23H1 in serum and cancer tissue and the relationship between them and clinicopathologic features in patients with gastric cancer.MethodsEmploying immunohistochemical S-P method and ELISA,the expressions of nm23H1 in serum and cancer tissue in 69 patients with gastric cancer were detected,and their correlation with clinicopathologic characteristics analyzed.ResultsThe expressions of nm23H1 in serum and cancer tissue in the patients were significantly different as compared with the normal control group(χ2＝9.69,t＝6.96,P＜0.05),the expressions were related with lymph-node metastasis and clinical staging(χ2＝12.30,5.30;t＝14.67,6.42;P＜0.05).ConclusionThe expressions of nm23H1 in serum and cancer tissue are down-regulated,and,therefore,nm23H1 can be used as an index for predicting prognosis of patients with gastric cancer.

stomach neoplasms;nm23H1;prognosis

R735.2

A

1008-0341(2014)01-0006-03

10.13362/j.qlyx201401003

2013-10-12;

2013-11-26

宋培鐸(1972-),男,碩士,主治醫(yī)師。