廣西右江民族醫(yī)學(xué)院附屬醫(yī)院，廣西 百色533000

結(jié)直腸癌患者血清中可溶性P-選擇素和L-選擇素水平及臨床意義

張梁 羅桂飛 元輝雄 韋貴將

廣西右江民族醫(yī)學(xué)院附屬醫(yī)院，廣西 百色533000

背景與目的：結(jié)直腸癌是常見的消化系統(tǒng)惡性腫瘤，嚴(yán)重威脅著人類的健康。雖然現(xiàn)代診療技術(shù)不斷發(fā)展，但近年來結(jié)直腸癌的發(fā)病率和病死率仍呈逐年上升趨勢(shì)，早期診斷和防治肝轉(zhuǎn)移有助于提高結(jié)直腸癌患者的生存率，對(duì)改善其預(yù)后有重要意義。P-選擇素和L-選擇素是近年腫瘤研究領(lǐng)域倍受關(guān)注的分子，其表達(dá)量及介導(dǎo)的黏附作用的改變?cè)谀[瘤細(xì)胞轉(zhuǎn)移中起著重要作用，但2者與結(jié)直腸癌發(fā)生及臨床分期的關(guān)系鮮見報(bào)道。本研究旨在探討結(jié)直腸癌患者手術(shù)前后血清中P-選擇素和L-選擇素水平高低與臨床病理特征的相關(guān)性。方法：選擇132例結(jié)直腸癌患者及與其性別年齡相匹配的100名健康體檢者。采用酶聯(lián)免疫吸附試驗(yàn)(enzyme-linked immunosorbent assay，ELISA)檢測(cè)132例結(jié)直腸癌患者手術(shù)前后血清中P-選擇素和L-選擇素濃度，100名健康體檢者為對(duì)照組，結(jié)合臨床病理特征對(duì)P-選擇素和L-選擇素水平的變化進(jìn)行分析。結(jié)果：結(jié)直腸癌患者術(shù)前血清P-選擇素和L-選擇素水平顯著高于對(duì)照組(75.2±13.3 vs 26.6±9.6，89.2±12.7 vs 33.9±8.3，P＜0.05)；術(shù)后1周P-選擇素和L-選擇素水平較術(shù)前沒有明顯下降(63.8±11.1 vs 75.2±13.3，71.5±10.9 vs 89.2±12.7，P＞0.05)；而術(shù)后3個(gè)月則明顯下降，與術(shù)前比較差異有統(tǒng)計(jì)學(xué)意義(34.8±10.4 vs 75.2±13.3，40.1±9.5 vs 89.2±12.7，P＜0.01)；本組結(jié)直腸癌患者，臨床分期越高，P-選擇素和L-選擇素水平越高，其水平與腫瘤浸潤(rùn)深度、Dukes分期、組織學(xué)分級(jí)和淋巴結(jié)轉(zhuǎn)移相關(guān)，而與年齡、性別及腫瘤部位無(wú)明顯相關(guān)(P＞0.05)；同時(shí)，同一結(jié)直腸癌患者血清中P-選擇素和L-選擇素的表達(dá)無(wú)顯著相關(guān)(r=4.114，P＞0.05)。結(jié)論：血清中P-選擇素和L-選擇素的水平在一定程度上反映了結(jié)直腸癌的侵襲、浸潤(rùn)和淋巴結(jié)轉(zhuǎn)移的程度，可作為預(yù)測(cè)結(jié)直腸癌發(fā)展及預(yù)后的重要指標(biāo)。

結(jié)直腸癌；P-選擇素；L-選擇素

結(jié)直腸癌是當(dāng)代常見的消化系統(tǒng)惡性腫瘤，嚴(yán)重威脅著人類的健康［1］。雖然現(xiàn)代診療技術(shù)不斷發(fā)展，但近年來國(guó)內(nèi)結(jié)直腸癌的發(fā)病率和病死率仍呈逐年上升趨勢(shì)［2］。早期診斷和防治肝轉(zhuǎn)移有助于提高結(jié)直腸癌患者的生存率，對(duì)改善其預(yù)后有重要意義［2］。而結(jié)直腸癌的浸潤(rùn)、轉(zhuǎn)移和術(shù)后復(fù)發(fā)是預(yù)后不良的主要原因，其發(fā)生、發(fā)展是多因素、多步驟、多分子參與的復(fù)雜過程［3-5］。隨著腫瘤因子的深入研究，結(jié)直腸癌相關(guān)因子的定位和識(shí)別已成為研究的熱點(diǎn)［6］。P-選擇素和L-選擇素是近年來腫瘤研究領(lǐng)域倍受關(guān)注的分子，其介導(dǎo)的黏附作用的改變?cè)谀[瘤細(xì)胞轉(zhuǎn)移中起著重要作用［7-9］，但2者與結(jié)直腸癌發(fā)生及臨床分期的關(guān)系鮮見報(bào)道。2008年3月—2011年3月，我們選擇了132例結(jié)直腸癌患者及與其性別年齡相匹配的100例健康體檢者。采用酶聯(lián)免疫吸附試驗(yàn)(enzyme-linked immunosorbent assay，ELISA)檢測(cè)132例結(jié)直腸癌患者手術(shù)前后血清中P-選擇素和L-選擇素濃度，100名健康體檢者為對(duì)照組，結(jié)合臨床病理特征對(duì)P-選擇素和L-選擇素水平的變化進(jìn)行分析，為P-選擇素和L-選擇素作為預(yù)測(cè)結(jié)直腸癌發(fā)生、發(fā)展及預(yù)后指標(biāo)提供分子實(shí)驗(yàn)依據(jù)。

1 資料和方法

1.1 臨床資料

選擇廣西右江民族醫(yī)學(xué)院附屬醫(yī)院于2008年3月—2011年3月收治的結(jié)直腸癌患者132例，診斷依據(jù)為臨床癥狀、B超、X線、CT檢查、腸道檢查及病理學(xué)診斷。其中男性83例、女性49例；年齡43～71歲，平均年齡64歲；浸潤(rùn)局限于漿膜內(nèi)50例，浸及漿膜外82例；臨床Dukes分期，Ⅰ期19例，Ⅱ期27例，Ⅲ期56例，Ⅳ期30例；組織學(xué)分級(jí)，高分化31例，中分化53例，低分化48例；無(wú)淋巴結(jié)轉(zhuǎn)移53例，有淋巴結(jié)轉(zhuǎn)移79例。對(duì)照組為100名來自本院體檢中心的健康體檢者，其中男性70例，女性30例，年齡38～75歲，平均年齡60歲，體溫、胸片、血尿常規(guī)正常，既往無(wú)心、肝、肺及腎等臟器疾病。所有患者術(shù)前未采用任何放化療或生物治療。

1.2 檢測(cè)方法

實(shí)驗(yàn)所需血清樣本均為清晨空腹靜脈血(3 mL)，3 000 r/min離心10 min，收集上層血清，采用ELISA法檢測(cè)血清中P-選擇素和L-選擇素水平，試劑盒為美國(guó)Gene May公司產(chǎn)品，實(shí)驗(yàn)步驟嚴(yán)格按照說明書進(jìn)行。術(shù)后1周及術(shù)后3個(gè)月采血及檢測(cè)方法相同。

1.3 統(tǒng)計(jì)學(xué)處理

采用SPSS 17.0統(tǒng)計(jì)軟件進(jìn)行數(shù)據(jù)分析，計(jì)量資料數(shù)據(jù)采用記錄，用t檢驗(yàn)，組間分析采用列變量為有序變量的行×列表的χ2檢驗(yàn)。P＜0.05為差異有統(tǒng)計(jì)學(xué)意義。

2 結(jié) 果

2.1 結(jié)直腸癌組和正常對(duì)照組血清P-選擇素和L-選擇素水平檢測(cè)結(jié)果比較

132例結(jié)直腸癌患者血清P-選擇素和L-選擇素水平顯著高于正常對(duì)照組(P＜0.05)；術(shù)后1周與術(shù)前血清水平變化差異不大(P＞0.05)，而與正常組比較差異有統(tǒng)計(jì)學(xué)意義(P＜0.05)；術(shù)后3個(gè)月與術(shù)前血清水平變化差異有統(tǒng)計(jì)學(xué)意義(P＜0.05)，與正常組比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05，表1)。

2.2 P-選擇素和L-選擇素水平與結(jié)直腸癌臨床病理特征的關(guān)系

以年齡、性別、腫瘤部位、浸潤(rùn)深度、Dukes分期、組織學(xué)分級(jí)及淋巴結(jié)轉(zhuǎn)移為指標(biāo)，對(duì)132例結(jié)直腸癌患者分別獨(dú)立進(jìn)行分類，研究P-選擇素和L-選擇素在各個(gè)分類組間的表達(dá)差異，發(fā)現(xiàn)P-選擇素和L-選擇素水平在結(jié)直腸癌患者血清中的表達(dá)與患者的年齡、性別、腫瘤部位差異無(wú)統(tǒng)計(jì)學(xué)意義(P＞0.05)，而與浸潤(rùn)深度、Dukes分期、組織學(xué)分級(jí)及淋巴結(jié)轉(zhuǎn)移差異有統(tǒng)計(jì)學(xué)意義(P＜0.05，表2)。

表1 結(jié)直腸癌組與正常對(duì)照組血清P-選擇素和L-選擇素水平檢測(cè)結(jié)果比較Tab. 1 The test results of serum P-selectin and L-selectin levels in colorectal cancer and normal control group

表2 結(jié)直腸癌患者血清中P-選擇素和L-選擇素的表達(dá)與臨床病理特征的關(guān)系Tab. 2 The relationship between the P-selectin and L-selectin expression of colorectal cancer patients and the clinical and pathological features

2.3 結(jié)直腸癌患者血清中P-選擇素和L-選擇素表達(dá)的相關(guān)性

將132例患者P-選擇素和L-選擇素有無(wú)表達(dá)進(jìn)行計(jì)數(shù)分類，采用列變量為有序變量的行×列表的χ2檢驗(yàn)，檢驗(yàn)P-選擇素和L-選擇素表達(dá)有無(wú)關(guān)聯(lián)。相關(guān)分析顯示P-選擇素和L-選擇素表達(dá)無(wú)顯著相關(guān)(χ2=4.114，P＞0.05，表3)。

表3 結(jié)直腸癌患者血清中P-選擇素和L-選擇素表達(dá)的關(guān)系Tab. 3 Relations P-selectin and L-selectin in serum of patients with colorectal cancer

3 討 論

可溶性P-選擇素和L-選擇素屬于選擇素家族的功能性細(xì)胞黏附分子，僅存于活化內(nèi)皮細(xì)胞表面［9］。正常組織中的內(nèi)皮細(xì)胞一般不表達(dá)P-選擇素和L-選擇素，只有當(dāng)內(nèi)皮細(xì)胞受到某些炎性因子如細(xì)菌脂多糖、白細(xì)胞介素、腫瘤壞死因子等刺激被激活時(shí)才表達(dá)，是炎性反應(yīng)過程中最早出現(xiàn)的黏附分子［10］。P-選擇素和L-選擇素介導(dǎo)活化的內(nèi)皮細(xì)胞與中性粒細(xì)胞的黏附作用，有助于白細(xì)胞穩(wěn)定地黏附于內(nèi)皮細(xì)胞，繼而遷移至血管外組織，從而促進(jìn)腫瘤細(xì)胞的侵襲轉(zhuǎn)移［11-12］。

腫瘤的侵襲轉(zhuǎn)移是一個(gè)多步驟、多環(huán)節(jié)、多因素、多分子參與的復(fù)雜過程，在這一過程中，多種分子各自發(fā)揮不同作用，并在時(shí)間和空間上相互配合，協(xié)同促進(jìn)細(xì)胞的癌變。近年來，隨著分子生物學(xué)發(fā)展，對(duì)P-選擇素和L-選擇素的結(jié)構(gòu)、功能、調(diào)控及病理生理作用進(jìn)行了較為深入的研究。查閱文獻(xiàn)發(fā)現(xiàn)［13］，P-選擇素和L-選擇素的表達(dá)受體內(nèi)外許多調(diào)節(jié)因子的影響，靜息時(shí)，內(nèi)皮細(xì)胞上的P-選擇素和L-選擇素含量甚微，當(dāng)受到炎性反應(yīng)因子刺激后，P-選擇素和L-選擇素表達(dá)顯著增加。有研究證實(shí)［14-15］，P-選擇素和L-選擇素與肺癌、膠質(zhì)瘤、胃癌、卵巢癌和腎癌等的浸潤(rùn)轉(zhuǎn)移的有關(guān)。Zheng等［16］發(fā)現(xiàn)P-選擇素的表達(dá)水平與肝癌患者的預(yù)后關(guān)系十分密切，P-選擇素的表達(dá)水平越低，其5年生存率越高；Cooney等［17］發(fā)現(xiàn)乳腺癌淋巴轉(zhuǎn)移患者中，P-選擇素在原位灶、轉(zhuǎn)移灶的血管內(nèi)皮和淋巴細(xì)胞中的高表達(dá)與腫瘤的高轉(zhuǎn)移有關(guān)；Radhakrishnan等［18］還發(fā)現(xiàn)前列腺癌患者血清中L-選擇素水平異常升高。我們的研究顯示，結(jié)直腸癌患者血清P-選擇素和L-選擇素水平明顯高于正常對(duì)照組，且與腫瘤的浸潤(rùn)、Dukes分期、有無(wú)淋巴結(jié)轉(zhuǎn)移相關(guān)，患者術(shù)后3個(gè)月血清P-選擇素和L-選擇素水平比術(shù)前明顯降低，可能提示P-選擇素和L-選擇素參與了結(jié)直腸癌的發(fā)展、侵襲和轉(zhuǎn)移。本研究還發(fā)現(xiàn)，術(shù)后1周血清P-選擇素和L-選擇素水平與術(shù)前相比無(wú)明顯變化，可能是患者仍處于應(yīng)激期，機(jī)體內(nèi)炎性反應(yīng)過程尚未消退，手術(shù)創(chuàng)傷刺激大量炎性因子分泌，使得血清中P-選擇素和L-選擇素仍維持在高水平。進(jìn)一步分析P-選擇素和L-選擇素表達(dá)與腫瘤臨床病理因素的相關(guān)性發(fā)現(xiàn)，P-選擇素和L-選擇素水平與患者的年齡、性別、腫瘤部位無(wú)關(guān)。但與癌細(xì)胞浸潤(rùn)的深度及Dukes分期、組織學(xué)分級(jí)、臨床分級(jí)及有無(wú)淋巴結(jié)轉(zhuǎn)移相關(guān)。提示密切觀察病情，動(dòng)態(tài)檢測(cè)患者血清P-選擇素和L-選擇素水平變化對(duì)預(yù)測(cè)結(jié)直腸癌轉(zhuǎn)移，觀察病情、判斷療效及評(píng)估預(yù)后具有重要意義。研究結(jié)果還顯示，P-選擇素和L-選擇素表達(dá)在結(jié)直腸癌患者中無(wú)相關(guān)關(guān)系，是2個(gè)各自發(fā)揮獨(dú)立作用的調(diào)控因子。

總之，P-選擇素和L-選擇素在結(jié)直腸癌的發(fā)生、發(fā)展、浸潤(rùn)轉(zhuǎn)移過程中均發(fā)揮著各自重要的作用，其表達(dá)水平能夠反映結(jié)直腸癌患者的預(yù)后狀況，也可能成為1項(xiàng)新的腫瘤標(biāo)志物，深入研究和監(jiān)測(cè)P-選擇素和L-選擇素將有助于判斷結(jié)直腸癌的發(fā)展趨勢(shì)和預(yù)后評(píng)估。

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Soluble P-selectin and L-selectin levels in serum of colorectal cancer patients and its clinical significance

ZHANG Liang, LUO Gui-fei, YUAN Hui-xiong, WEI Gui-jiang
(Affiliated Hospital of

Youjiang Medical University for Nationalities, Baise Guangxi 533000, China)

LUO Gui-fei E-mail: luoguifei2012@163.com

Background and purpose: Colorectal cancer is a kind of common digestive malignancies, which seriously threaten the human health. Although modern diagnostic and treatment technology has developed rapidly, the incidence and mortality of colorectal cancer continue to show an increasing tendency in recent years, and early diagnosis and prevention of colorectal cancer liver metastases are important to increase the survival rate of patients and to improve the prognosis. P-selectin and L-selectin is attracting much attention in cancer research field recently, and the change of their expression and mediated adhesion play an important role in tumor cell metastasis, but their relationship between the occurrence and clinical stage of colorectal cancer rarely reported. This study aimed to explore whether the serum P-selectin and L-selectin levels of colorectal cancer patients were correlated with clinical and pathological features and the situation before and after surgery. Methods: A total number of 132 cases of colorectal cancer patients and 100 healthy subjects with gender and age-matched were enrolled. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum P-selectin and L-selectin concentrations in 132 cases before and after surgery, 100 healthy subjects were enrolled as the control group, the P-selectin and L-selectin levels were analyzed combined with theclinical and pathological features. Results: Serum P-selectin and L-selectin levels in patients with colorectal cancer before surgery were signi ficantly higher than those in the healthy control group (75.2±13.3 vs 26.6±9.6, 89.2±12.7 vs 33.9±8.3, P＜0.05) ; Compared with the patients before surgery, 1 week after surgery, the P-selectin and L-selectin levels were not significantly decreased (63.8±11.1 vs 75.2±13.3, 71.5±10.9 vs 89.2±12.7, P＞0.05); however, significantly decreased after 3 months (34.8±10.4 vs 75.2±13.3, 40.1±9.5 vs 89.2±12.7, P＜0.01); This paper studied patients with colorectal cancer, the higher clinical stage, higher P-selectin and L-selectin levels, and their expression levels with tumor invasion depth, Dukes stage, histological grade and lymph node metastasis were related, but were not correlated with age, gender and tumor site. Meanwhile, P-selectin and L-selectin levels were not signi ficantly correlated in the same serum of patients with colorectal cancer (r=4.114, P＞0.05). Conclusion: Serum P-selectin and L-selectin levels to some extent re fl ect the invasion of colorectal cancer, the degree of in filtration and lymph node metastasis, and which can be an important indicator in the development and prognosis of colorectal cancer.

Colorectal cancer; P-selectin; L-selectin

10.3969/j.issn.1007-3969.2014.08.007

R735.3

A

1007-3639(2014)08-0599-05

2013-12-02

2014-05-17）

廣西衛(wèi)生廳資助項(xiàng)目(No：百科計(jì)字［2010］5號(hào))。

羅桂飛 E-mail:luoguifei2012@163.com