口腔癌患者外周血細(xì)胞免疫水平與臨床結(jié)局的相關(guān)性分析

孟智堅，郭立娜

口腔癌患者外周血細(xì)胞免疫水平與臨床結(jié)局的相關(guān)性分析

孟智堅，郭立娜

目的 探討口腔癌患者外周血細(xì)胞免疫水平與臨床結(jié)局的相關(guān)性。方法 選取2014年1月—2015年1月我院收治的口腔癌50例(口腔癌組)，選擇同期體檢健康的50例(對照組)。比較兩組外周血淋巴細(xì)胞(CD3+、CD4+、CD8+、CD19+)和自然殺傷(natural killer, NK)細(xì)胞的水平，分析口腔癌患者外周血淋巴細(xì)胞(CD3+、CD4+、CD8+、CD19+)、NK細(xì)胞與其臨床特征的相關(guān)性。結(jié)果 與對照組比較，口腔癌組CD3+及CD4+T淋巴細(xì)胞、CD4+/CD8+和NK細(xì)胞顯著降低(P<0.05)，CD19+T淋巴細(xì)胞顯著升高 (P=0.000)，CD8+T淋巴細(xì)胞降低不明顯(P>0.05)。與Ⅲ～Ⅳ期口腔癌患者比較，Ⅰ～Ⅱ期患者的CD3+、CD4+及CD8+T淋巴細(xì)胞、CD4+/CD8+和NK細(xì)胞顯著升高(P<0.05)，CD19+T淋巴細(xì)胞顯著降低(P=0.000)。與無淋巴結(jié)轉(zhuǎn)移的患者比較，出現(xiàn)淋巴結(jié)轉(zhuǎn)移的患者CD3+、CD4+及CD8+T淋巴細(xì)胞和NK細(xì)胞顯著降低(P<0.05)，CD19+T淋巴細(xì)胞顯著升高(P=0.000)，CD4+/CD8+降低不明顯(P>0.05)。與無復(fù)發(fā)患者比較，術(shù)后復(fù)發(fā)者CD3+、CD4+及CD8+T淋巴細(xì)胞、CD4+/CD8+和NK細(xì)胞顯著降低(P<0.05)，CD19+T淋巴細(xì)胞顯著升高(P=0.000)。結(jié)論 口腔癌患者存在一定程度的免疫抑制，且外周血細(xì)胞免疫水平與其預(yù)后密切相關(guān)。

口腔腫瘤；免疫，細(xì)胞；T淋巴細(xì)胞

近年來隨著環(huán)境的污染和人類生活習(xí)慣的改變，口腔癌的發(fā)病率逐年升高[1]。研究顯示惡性腫瘤患者常伴有不同程度的免疫功能障礙[2-3]，其中外周血細(xì)胞免疫功能障礙較常見。T淋巴細(xì)胞和自然殺傷 (natural killer, NK)細(xì)胞是外周免疫系統(tǒng)的重要組成成分，在免疫介導(dǎo)的抗腫瘤反應(yīng)中發(fā)揮關(guān)鍵性作用[4-5]。文獻(xiàn)報道腫瘤患者存在一定程度的輔助性T細(xì)胞(Th)1/Th2平衡失調(diào)，其可進(jìn)一步促進(jìn)腫瘤細(xì)胞的增殖和轉(zhuǎn)移，故腫瘤患者免疫功能低下時常伴病情惡化[6-7]。因此，探討口腔癌患者的細(xì)胞免疫功能具有十分重要的臨床意義，可為進(jìn)一步的臨床干預(yù)提供理論依據(jù)，但國內(nèi)相關(guān)研究較少。本文探討口腔癌患者外周血細(xì)胞免疫水平與臨床結(jié)局的相關(guān)性，現(xiàn)將結(jié)果報告如下。

1 資料與方法

1.1 一般資料 選取我院2014年1月—2015年1月收治的口腔癌50例(口腔癌組)，其中男31例，女19例；年齡29～63(49.93±12.39)歲；患中舌癌18例，口底癌12例，牙齦癌9例，頰癌7例，上顎癌4例；腫瘤細(xì)胞高分化28例，中分化14例，低分化8例；TNM分期為Ⅰ期11例，Ⅱ期9例，Ⅲ期23例，Ⅳ期7例；合并淋巴結(jié)轉(zhuǎn)移39例，合并遠(yuǎn)處轉(zhuǎn)移7例。納入標(biāo)準(zhǔn)：①經(jīng)病理檢查確診的口腔癌患者；②入院前未接受過放化療、手術(shù)等治療；③年齡18～65歲；④同意參與本研究。排除標(biāo)準(zhǔn)：①復(fù)發(fā)的口腔癌；②伴全身系統(tǒng)性疾病；③多臟器功能不全；④5年內(nèi)合并其他腫瘤；⑤伴免疫功能缺陷；⑥不配合治療或治療期間轉(zhuǎn)院；⑦隨訪期間失訪。選取同期在我院體檢健康的50例作為對照組，其中男31例，女19例；年齡28～63(50.18±12.73)歲。兩組的一般資料比較差異無統(tǒng)計學(xué)意義(P>0.05)，具有可比性。本研究通過我院倫理委員會批準(zhǔn)，所有研究對象均知情同意并簽署知情同意書。

1.2 研究方法 兩組均于清晨空腹抽取靜脈血5 ml，3000 r/min離心10 min后取上層血清，置于乙二胺四乙酸二鈉試管中，抽取100 μl與鼠抗人單克隆抗體CD45/CD4/CD8/CD3、CD45/CD(16+56)/CD19/CD3各20 μl混勻，常溫避光下反應(yīng)30 min，于60 min內(nèi)進(jìn)行流式細(xì)胞儀檢測(Epics XL型流式細(xì)胞儀，美國Beckman coulter公司)，測定血淋巴細(xì)胞CD3+、CD4+、CD8+、CD19+和NK細(xì)胞的水平。

1.3 觀察指標(biāo) 比較兩組外周血淋巴細(xì)胞(CD3+、CD4+、CD8+、CD19+)和NK細(xì)胞的水平變化，分析口腔癌患者外周血淋巴細(xì)胞(CD3+、CD4+、CD8+、CD19+)及 NK細(xì)胞與其臨床特征的相關(guān)性。

2 結(jié)果

2.1 兩組外周血淋巴細(xì)胞水平比較 與對照組比較，口腔癌組CD3+及CD4+T淋巴細(xì)胞、CD4+/CD8+和NK細(xì)胞顯著降低(P<0.05)，CD19+T淋巴細(xì)胞顯著升高 (P=0.000)，CD8+T淋巴細(xì)胞降低不明顯(P>0.05)，見表1。

2.2 TNM分期與外周血淋巴細(xì)胞的相關(guān)性 與Ⅲ～Ⅳ期口腔癌患者比較，Ⅰ～Ⅱ期患者的CD3+、CD4+及CD8+T淋巴細(xì)胞、CD4+/CD8+和NK細(xì)胞顯著升高(P<0.05)，CD19+T淋巴細(xì)胞顯著降低(P=0.000)，見表2。

表1 口腔癌患者與健康對照組外周血淋巴細(xì)胞水平比較

表2 口腔癌患者TNM分期與外周血淋巴細(xì)胞的相關(guān)性

2.3 淋巴結(jié)有無轉(zhuǎn)移與外周血淋巴細(xì)胞的相關(guān)性 與無淋巴結(jié)轉(zhuǎn)移的患者比較，出現(xiàn)淋巴結(jié)轉(zhuǎn)移的患者CD3+、CD4+及CD8+T淋巴細(xì)胞和NK細(xì)胞顯著降低(P<0.05)，CD19+T淋巴細(xì)胞顯著升高(P=0.000)，CD4+/CD8+降低不明顯(P>0.05)，見表3。

表3 口腔癌患者有無淋巴結(jié)轉(zhuǎn)移與外周血淋巴細(xì)胞的相關(guān)性

2.4 術(shù)后有無復(fù)發(fā)與外周血淋巴細(xì)胞的相關(guān)性 本組43例行根治性手術(shù)治療，隨訪1年后15例復(fù)發(fā)。與無復(fù)發(fā)患者比較，術(shù)后復(fù)發(fā)者CD3+、CD4+及CD8+T淋巴細(xì)胞、CD4+/CD8+和NK細(xì)胞顯著降低(P<0.05)，CD19+T淋巴細(xì)胞顯著升高(P=0.000)，見表4。

表4 口腔癌患者行根治性手術(shù)后有無復(fù)發(fā)與外周血淋巴細(xì)胞的相關(guān)性

3 討論

以T淋巴細(xì)胞和NK細(xì)胞為主的細(xì)胞免疫在機(jī)體抗腫瘤中發(fā)揮著重大作用。CD8+T淋巴細(xì)胞是細(xì)胞免疫的重要組成，CD3+T淋巴細(xì)胞則是成熟T淋巴細(xì)胞的標(biāo)志物[8]，均在細(xì)胞免疫和體液免疫中起著關(guān)鍵性協(xié)助作用，而CD19+T淋巴細(xì)胞在細(xì)胞免疫功能受損時，反應(yīng)性增高[9]。我國學(xué)者發(fā)現(xiàn)組織缺氧可通過誘導(dǎo)CCL28的高表達(dá)，進(jìn)而促進(jìn)調(diào)節(jié)性T淋巴細(xì)胞聚集，促進(jìn)原發(fā)性肝癌患者的腫瘤細(xì)胞生長[10]。Pedroza-Gonzalez等[11]研究顯示原發(fā)性或轉(zhuǎn)移性肝癌患者腫瘤微環(huán)境中CD4+細(xì)胞源的調(diào)節(jié)性T淋巴細(xì)胞增加，使機(jī)體的免疫力降低，與預(yù)后呈正相關(guān)。Li等[12]研究表明外周血調(diào)節(jié)性T淋巴細(xì)胞水平的升高與臨床預(yù)后有關(guān)。鑒于免疫系統(tǒng)在腫瘤發(fā)生和發(fā)展中的重要意義，急需探討口腔癌患者免疫功能與臨床預(yù)后的相關(guān)性，但目前相關(guān)研究尚未完善。

本文系統(tǒng)探討了口腔癌患者CD3+、CD4+、CD8+及CD19+T淋巴細(xì)胞和NK細(xì)胞的表達(dá)，在很大程度上反映了患者的免疫功能。本研究結(jié)果顯示，與對照組比較，口腔癌組CD3+及CD4+T淋巴細(xì)胞、CD4+/CD8+及NK細(xì)胞顯著降低，CD19+T細(xì)胞顯著升高，表明口腔癌患者免疫功能受損。此外，與Ⅲ～Ⅳ期口腔癌患者比較，Ⅰ～Ⅱ期患者的CD3+、CD4+及CD8+T淋巴細(xì)胞和CD4+/CD8+、NK細(xì)胞顯著升高，CD19+T淋巴細(xì)胞顯著降低；與無淋巴結(jié)轉(zhuǎn)移的患者比較，出現(xiàn)淋巴結(jié)轉(zhuǎn)移的患者CD3+、CD4+及CD8+T淋巴細(xì)胞和NK細(xì)胞顯著降低，CD19+T淋巴細(xì)胞顯著升高；與術(shù)后無復(fù)發(fā)的患者比較，術(shù)后復(fù)發(fā)者的CD3+、CD4+及CD8+T淋巴細(xì)胞、CD4+/CD8+和NK細(xì)胞顯著降低，CD19+T淋巴細(xì)胞顯著升高，說明口腔癌患者的病情越重，免疫功能越低下，與符良斌等[13]研究結(jié)果一致。T淋巴細(xì)胞亞群介導(dǎo)的細(xì)胞免疫和NK細(xì)胞可發(fā)揮殺傷腫瘤細(xì)胞的作用，其功能抑制可能與患者術(shù)后復(fù)發(fā)率升高有關(guān)，且免疫功能低下是發(fā)生腫瘤的重要因素，表現(xiàn)為輔助性T淋巴細(xì)胞減少而抑制性T淋巴細(xì)胞增多，腫瘤切除或許可解除患者的免疫抑制狀態(tài)[14]。張立娜[15]研究顯示口腔癌患者外周血調(diào)節(jié)性T淋巴細(xì)胞表達(dá)水平升高，手術(shù)切除原發(fā)病灶后有助于降低外周血調(diào)節(jié)性T淋巴細(xì)胞水平，進(jìn)而解除免疫抑制狀態(tài)。

綜上所述，口腔癌患者存在一定程度的免疫抑制，外周血細(xì)胞免疫水平與臨床預(yù)后密切相關(guān)。

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Correlation Analysis of Immune Cell Levels of Peripheral Blood and Clinical Outcomes in Patients with Mouth Cancer

MENG Zhi-jian, GUO Li-na

(Department of Stomatology, Affiliated Hospital of Inner Mongolia Medical University, Hohhot 010050, China)

Objective To investigate correlation between immune cell levels of peripheral blood and clinical outcomes in patients with month cancer. Methods A total of 50 patients with month cancer admitted during January 2014 and January 2015 were selected as month cancer group, and 50 healthy people taking medical examination were chosen as control group. Levels of peripheral blood lymphocytes (CD3+, CD4+, CD8+and CD19+) and natural killer (NK) cells were compared, and correlations between levels of peripheral blood lymphocytes (CD3+, CD4+, CD8+and CD19+) and NK cell and clinical characteristics were analyzed in two groups. Results Compared with those in control group, levels of CD3+ and CD4+ T lymphocyte, CD4+/CD8+and NK lymphocyte were significantly decreased (P<0.05); CD19+ T lymphocyte level was significantly increased (P=0.000); while CD8+T lymphocyte level was not significantly decreased in month cancer group (P>0.05). Compared with those in patients with stage III-IV, levels of CD3+, CD4+and CD8+T lymphocyte, CD4+/CD8+and NK lymphocyte were significantly increased (P<0.05); CD19+T cell level was significantly decreased in patients with stage I-II (P=0.000). Compared with those in patients without lymph node metastasis, levels of CD3+, CD4+and CD8+T lymphocyte and NK cell level were significantly decreased (P<0.05), while CD19+T cell level was significantly increased (P=0.000), but CD4+/CD8+level was not significantly decreased in patients with lymph node metastasis (P>0.05). Compared with those in patients without recurrence, levels of CD3+CD4+and CD8+T lymphocyte, CD4+/CD8+and NK lymphocyte were significantly decreased (P<0.05) and CD19+T lymphocyte level was significantly increased in patients with postoperative recurrence (P=0.000). Conclusion Patients with month cancer have a certain degree of immune suppression, and immunity cell levels of peripheral blood is closely associated with clinical prognosis.

Mouth neoplasms; Immunity, cellular; T-Lymphocytes

內(nèi)蒙古自治區(qū)衛(wèi)生和計劃生育委員會醫(yī)療衛(wèi)生科研計劃項(xiàng)目(201303048)

010050 呼和浩特，內(nèi)蒙古醫(yī)科大學(xué)附屬醫(yī)院口腔科

郭立娜，E-mail:2649122035@qq.com

R739.8

A

1002-3429(2017)03-0074-04

10.3969/j.issn.1002-3429.2017.03.029

2016-11-29 修回時間：2016-12-29)