扶志敏 鄺世晏 李熾光 劉俊永 朱云 陳尚志
[摘要] 目的 明確非小細(xì)胞肺癌患者外周血細(xì)胞因子IL-2、IL-6、TNF-α和IFN-γ變化及與生存率的相互關(guān)系。方法 招募2016~2017年53例非小細(xì)胞肺癌住院患者和53例健康對(duì)照者作為研究對(duì)象。收集非小細(xì)胞肺癌組入院第7天、30 d、90 d、180 d血清,ELSIA法檢測(cè)并比較肺癌組和健康組血清中IL-2,IL-6,TNF-α和IFN-γ不同時(shí)間段表達(dá)差異。通過(guò)分析不同時(shí)間段非小細(xì)胞肺癌組血清中細(xì)胞因子與體重相互關(guān)系及COX回歸生存分析明確影響非小細(xì)胞肺癌生存率的細(xì)胞因子。 結(jié)果 與健康對(duì)照組比較,非小細(xì)胞肺癌組血清中細(xì)胞因子IL-6和IFN-γ在第7天、30 d、90 d、180 d均出現(xiàn)明顯升高(P<0.001,P<0.001,P<0.001,P<0.001和P=0.161,P<0.001,P=0.001,P=0.041);而IL-2,TNF-α水平隨住院時(shí)間則無(wú)明顯變化。進(jìn)一步分析發(fā)現(xiàn)非小細(xì)胞肺癌第90天和180天血清中IL-6與體重呈負(fù)相關(guān)(R=-0.368,P=0.045和R=-0.497,P=0.011)。COX回歸生存分析發(fā)現(xiàn)非小細(xì)胞肺癌組IL-6水平與患者生存率存在負(fù)相關(guān)(P=0.023)。 結(jié)論 非小細(xì)胞肺癌會(huì)引起血清中IL-6持續(xù)升高,持續(xù)升高的IL-6可與非小細(xì)胞肺癌患者的生存率減少密切相關(guān),血清中IL-6可作為非小細(xì)胞肺癌發(fā)生發(fā)展的生物標(biāo)記物之一。
[關(guān)鍵詞] 非小細(xì)胞肺癌;血清;IL-6;生存率
[中圖分類號(hào)] R730.4? ? ? ? ? [文獻(xiàn)標(biāo)識(shí)碼] A? ? ? ? ? [文章編號(hào)] 1673-9701(2019)11-0032-05
Correlation between changes of serum IL-2, IL-6, TNF-α and IFN-γ in serum of non-small cell lung cancer and survival rate
FU Zhimin1? ?KUANG Shiyan1? ?LI Chiguang2? ?LIU Junyong2? ?ZHU Yun1? ?CHEN Shangzhi3
1.Department of Cardiothoracic Surgery, Chenzhou First People's Hospital in Hu'nan Province, Chenzhou? ?423000, China; 2.Department of Respiratory Medicine, Chenzhou First People's Hospital in Hu'nan Province, Chenzhou? ?423000, China; 3.Office, Chenzhou First People's Hospital in Hu'nan Province, Chenzhou? ?423000, China
[Abstract] Objective To determine the changes of peripheral blood cytokines IL-2, IL-6, TNF-α and IFN-γ in patients with non-small cell lung cancer and their relationship with survival rate. Methods A total of 53 non-small cell lung cancer hospitalized patients and 53 healthy controls were enrolled from 2016 to 2017. The sera of the non-small cell lung cancer group were collected on the 7 th, 30 th, 90 th, and 180 th day of the hospital. ELSIA method was used to detect and compare the expression differences in serum IL-2, IL-6, TNF-α and IFN-γ in the lung cancer group and the healthy group over time. The cytokines affecting the survival rate of non-small cell lung cancer were identified by analyzing the relationship between cytokine and body weight in serum and COX regression survival analysis in non-small cell lung cancer group at different time points. Results Compared with those of the healthy control group, the serum cytokines IL-6 and IFN-γ in the non-small cell lung cancer group were significantly increased on the 7 th, 30 th, 90 th, and 180 th days (P<0.001, P<0.001, P<0.001, P<0.001 and P=0.116, P<0.001, P=0.001, P=0.041); while IL-2, TNF-α levels in the non-small cell lung cancer group did not change significantly with hospital stay. Further analysis found that serum IL-6 was negatively correlated with body weight on 90th days and 180th days of non-small cell lung cancer (R=-0.368, P=0.045 and R=-0.497, P=0.011). COX regression survival analysis found that IL-6 levels in non-small cell lung cancer group were negatively correlated with patient survival(P=0.023). Conclusion Non-small cell lung cancer can cause continuous increase of serum IL-6. The continuous increase of IL-6 is closely related to the decrease of survival rate in patients with non-small cell lung cancer. IL-6 in serum can be used as one of the biomarkers for the development of non-small cell lung cancer.
[Key words] Non-small cell lung cancer; Serum; IL-6; Survival rate
肺癌主要病理類型分為小細(xì)胞癌和非小細(xì)胞肺癌,其中非小細(xì)胞肺癌占總肺癌發(fā)病率的85%,是最主要的肺癌類型。非小細(xì)胞肺癌的主要組織學(xué)亞型是腺癌和鱗狀細(xì)胞癌。非小細(xì)胞肺癌預(yù)后較差,雖然小型局限性腫瘤(Ⅰ期)5年生存率為70%~90%,但是如果進(jìn)展到遠(yuǎn)處轉(zhuǎn)移(Ⅳ期)1年生存率僅為15%~19%[1],因此現(xiàn)階段非小細(xì)胞肺癌的早期發(fā)現(xiàn)和及時(shí)治療是提高患者生存率的重要手段。敏感度和特異度高的生物標(biāo)記物對(duì)非小細(xì)胞肺癌早期診斷具有重要意義,有研究提示外周血細(xì)胞因子的變化與腫瘤的侵襲及轉(zhuǎn)移存在密切聯(lián)系[2]。本研究通過(guò)分析非小細(xì)胞肺癌患者外周血細(xì)胞因子的變化與患者生存率之間的相互關(guān)系,探尋相關(guān)的生物標(biāo)志物,為肺癌的早期治療提供參考。
1 資料與方法
1.1 一般資料
2016年10月~2017年10月于我院招募53例非小細(xì)胞肺癌患者(排除其他腫瘤及高血壓、冠心病、糖尿病等慢性疾病和急慢性感染性疾病),其中男48例,女5例,年齡中位數(shù)59歲。同時(shí)選取我院53例健康體檢人群作為對(duì)照組,男43例,女10例,年齡中位數(shù)52歲。兩組人群在年齡和性別分布均無(wú)統(tǒng)計(jì)學(xué)差異(P=0.655和P=0.265)。見(jiàn)表1。
1.2 診斷及納入標(biāo)準(zhǔn)
參照中國(guó)抗癌協(xié)會(huì)腫瘤臨床化療專業(yè)委員會(huì)制定的《中國(guó)原發(fā)性肺癌診療規(guī)范》,對(duì)所有非小細(xì)胞肺癌患者進(jìn)行診斷及納入[3]。所有肺癌患者病理及臨床分期都是通過(guò)有經(jīng)驗(yàn)的病理和臨床醫(yī)生確定。
1.3 方法
1.3.1 樣本收集? 使用非抗凝采血管收集3 mL外周血,24 h內(nèi)3000 rpm離心 10 min后取血清凍存于-80℃冰箱。
1.3.2 血清中細(xì)胞因子的檢測(cè)? 血清中細(xì)胞因子使用酶聯(lián)免疫吸附法(Enzyme linked immunosorbent assay,ELISA)檢測(cè),Human IL-2 ELISA Kit,Human IL-6 ELISA Kit,Human TNF-α ELISA Kit,Human IFN-γ ELISA Kit購(gòu)于深圳欣博盛科技股份有限公司,所有操作均嚴(yán)格按照說(shuō)明書進(jìn)行。
1.4 統(tǒng)計(jì)學(xué)方法
兩組間比較Mann Whitney test檢驗(yàn),性別分布比較Fisher精確檢驗(yàn),兩變量之間的相互關(guān)系比較使用Spearman相關(guān)分析,肺癌患者生存分析使用COX回歸,P<0.05為差異有統(tǒng)計(jì)學(xué)意義。上述所有數(shù)據(jù)使用SPSS 19.0和Prism 6軟件進(jìn)行統(tǒng)計(jì)分析,失訪和死亡人群在兩組比較分析時(shí)被剔除,非小細(xì)胞肺癌組死亡人群納入到COX生存分析中。
2 結(jié)果
2.1 兩組一般臨床學(xué)特征比較
分析非小細(xì)胞肺癌患者的一般臨床學(xué)指標(biāo)如入院時(shí)的基本情況(7 d內(nèi)),與健康對(duì)照組相比,非小細(xì)胞肺癌組的外周淋巴細(xì)胞絕對(duì)數(shù)水平顯著上升(P<0.001),但其他臨床指標(biāo)并未見(jiàn)有統(tǒng)計(jì)學(xué)差異(P>0.05)。見(jiàn)表1。
2.2 血清中IL-2、IL-6、TNF-α、IFN-γ變化
與健康對(duì)照組相比,非小細(xì)胞肺癌組患者在入院后第7天、第30天、第90天及第180天其血清中的IL-6和IFN-γ水平顯著升高(P<0.001,P<0.001,P<0.001,P<0.001和P=0.161,P<0.001,P=0.001,P=0.041);IL-2、TNF-α水平與健康對(duì)照組比較無(wú)顯著變化(P>0.05)。見(jiàn)表2。
2.3 IL-6和IFN-γ變化與體重之間的關(guān)系
肺癌患者如果病情惡化往往會(huì)導(dǎo)致惡病質(zhì),其中最明顯的體征是體重的改變,因此我們將外周血中IL-6和IFN-γ的變化與患者體重的變化進(jìn)行相關(guān)分析,發(fā)現(xiàn)隨著非小細(xì)胞肺癌患者體重的下降往往伴隨著外周血IL-6上升,在第90天和第180天時(shí)IL-6血清水平與體重之間存在顯著性負(fù)相關(guān)(R=-0.368,P=0.045和R=-0.497,P=0.011)。而非小細(xì)胞肺癌患者體重與IFN-γ水平之間并未發(fā)現(xiàn)相關(guān)(P>0.05)。見(jiàn)圖1。由于體重是衡量癌變預(yù)后的重要指標(biāo)之一,因此上述結(jié)果提示血清中IL-6表達(dá)可能與非小細(xì)胞肺癌預(yù)后相關(guān)。
圖1? ?肺癌患者IL-6和IFN-γ變化與體重之間的關(guān)系
A.血清中IFN-γ水平和體重在不同時(shí)間點(diǎn)的變化情況;B.清中IL-6水平和體重在不同時(shí)間點(diǎn)的變化情況;C.第90天體重與血清中IL-6的相互關(guān)系;D.第180天體重與血清中IL-6的相互關(guān)系
2.4 血清IL-6與非小細(xì)胞肺癌生存分析
因?yàn)镮L-6在血清中存在明顯差異并且與患者體重負(fù)相關(guān),我們根據(jù)患者血清IL-6水平分為兩組:高水平組IL-6>6.4 pg/mL和低水平組IL-6<6.4 pg/mL。COX生存分析發(fā)現(xiàn)血清IL-6水平高組與IL-6低水平組相比較預(yù)后較差,生存率存在顯著的統(tǒng)計(jì)學(xué)差異(P=0.023),風(fēng)險(xiǎn)比(hazard ratio,HR)=3.732(95% CI:1.329~10.48)。見(jiàn)圖2。提示外周血中IL-6持續(xù)高表達(dá)可能與非小細(xì)胞肺癌的生存率有關(guān)。
圖2? ?非小細(xì)胞肺癌患者不同外周血IL-6水平組生存率分析
3 討論
細(xì)胞因子是炎癥反應(yīng)的主要介質(zhì),參與腫瘤免疫清除,并與腫瘤的預(yù)后有密切的關(guān)系。在腫瘤的發(fā)展過(guò)程中細(xì)胞因子不僅可以由機(jī)體的免疫系統(tǒng)產(chǎn)生來(lái)清除和抑制腫瘤生長(zhǎng),也可以由腫瘤細(xì)胞自身分泌產(chǎn)生來(lái)支持腫瘤自身的生長(zhǎng)及轉(zhuǎn)移。大量的研究表明炎癥因子與腫瘤發(fā)生發(fā)展密切相關(guān),如TNF-α的產(chǎn)生與肝癌風(fēng)險(xiǎn)增加相關(guān)[4],外周血清IL-6與卵巢細(xì)胞癌[5]、多發(fā)性骨髓瘤[6]、淋巴瘤[7]、膠質(zhì)母細(xì)胞瘤[8]的惡性程度呈正相關(guān),TGF-β參與促進(jìn)上皮腫瘤分化[9]。本研究分析了非小細(xì)胞肺癌患者外周血中IL-2、IL-6、TNF-α和IFN-γ變化與非小細(xì)胞肺癌病程之間的關(guān)系,發(fā)現(xiàn)血清中IL-6水平變化與非小細(xì)胞肺癌患者的生存率下降密切相關(guān),提示IL-6可能參與了非小細(xì)胞肺癌疾病的進(jìn)展。
IL-6屬于白細(xì)胞介素的一種細(xì)胞因子,主要由纖維母細(xì)胞、單核/巨噬細(xì)胞、T淋巴細(xì)胞、B淋巴細(xì)胞分泌。但是IL-6也可由多種瘤細(xì)胞所產(chǎn)生,如急性粒細(xì)胞白血病細(xì)胞[10]、口腔鱗癌細(xì)胞[11]和肺癌細(xì)胞等[12]均可以自發(fā)分泌IL-6。IL-6在腫瘤的發(fā)生發(fā)展中發(fā)揮的作用不同:一方面IL-6作為一種炎癥因子參與免疫反應(yīng)抑制腫瘤的生長(zhǎng),如IL-6通過(guò)激活Stat3信號(hào)通路降低雌激素和雄激素受體(ER和AR)的表達(dá),引起ER和AR陽(yáng)性癌細(xì)胞的生長(zhǎng)減少,從而抑制ER和AR陽(yáng)性相關(guān)腫瘤的生長(zhǎng)[13-15];另一方面IL-6參與很多腫瘤的轉(zhuǎn)移及生長(zhǎng),如腫瘤干細(xì)胞產(chǎn)生的IL-6可以自分泌方式促進(jìn)腫瘤細(xì)胞的存活和增殖[16]。IL-6除了能提高腫瘤干細(xì)胞的存活率外,還可以增加腫瘤細(xì)胞遷移并誘導(dǎo)與上皮-間質(zhì)細(xì)胞轉(zhuǎn)化[17-18]。動(dòng)物實(shí)驗(yàn)發(fā)現(xiàn)IL-6過(guò)表達(dá)的腫瘤細(xì)胞較IL-6敲除的腫瘤細(xì)胞更能促進(jìn)腫瘤細(xì)胞的轉(zhuǎn)移,而IL-6之所以能促進(jìn)腫瘤細(xì)胞生長(zhǎng)及轉(zhuǎn)移可能與過(guò)快的腫瘤生長(zhǎng)造成局部缺氧環(huán)境密切相關(guān)[19-20]。有研究發(fā)現(xiàn)缺氧能誘導(dǎo)IL-6的表達(dá)和信號(hào)傳遞,并且骨髓等缺氧部位是轉(zhuǎn)移細(xì)胞最常見(jiàn)的部位,化療和放射治療導(dǎo)致局部組織缺氧環(huán)境也是引起IL-6高表達(dá)而利于腫瘤細(xì)胞生長(zhǎng)的重要原因[21]。IL-6促進(jìn)腫瘤發(fā)生發(fā)展的分子學(xué)機(jī)制可能是通過(guò)Janus酪氨酸家族激酶(JAK)途徑促進(jìn)腫瘤細(xì)胞增殖[22],并上調(diào)血管內(nèi)皮生長(zhǎng)因子(VEGF)的表達(dá),促進(jìn)腫瘤血管生成及腫瘤的轉(zhuǎn)移[23]。因此IL-6水平與腫瘤的發(fā)生發(fā)展存在密切的關(guān)系。我們的研究發(fā)現(xiàn),外周血中IL-6高水平表達(dá)的非小細(xì)胞肺癌患者的生存率更低,這結(jié)果提示IL-6可能參與到非小細(xì)胞肺癌惡性化發(fā)展的過(guò)程中。
我們另一個(gè)發(fā)現(xiàn)是IL-6與非小細(xì)胞肺癌患者的體重呈負(fù)相關(guān),因?yàn)槟[瘤生長(zhǎng)需要大量維持機(jī)體的營(yíng)養(yǎng)物質(zhì),晚期腫瘤患者病情惡化往往會(huì)導(dǎo)致惡病質(zhì)的發(fā)生,所有體重是衡量腫瘤患者疾病是否惡性化的重要標(biāo)志。有研究發(fā)現(xiàn)IL-6可能誘導(dǎo)生長(zhǎng)激素釋放增加,而生長(zhǎng)激素水平增強(qiáng)會(huì)導(dǎo)致脂肪分解[24],腫瘤的發(fā)生往往伴隨著高代謝,而IL-6又參與了高代謝的過(guò)程,因此非小細(xì)胞肺癌患者的體重減輕可能與體內(nèi)IL-6異常表達(dá)存在密切相關(guān)。
綜上所述,非小細(xì)胞肺癌患者血清IL-6水平升高可能與其生存率下降密切相關(guān)。IL-6可能不僅參與非小細(xì)胞肺癌細(xì)胞的生長(zhǎng)及轉(zhuǎn)移,更可能參與到機(jī)體的基礎(chǔ)代謝消耗中,從而導(dǎo)致整個(gè)機(jī)體防御能力下降影響患者的生存率。本研究的發(fā)現(xiàn)提示血清IL-6可能作為非小細(xì)胞肺癌發(fā)生發(fā)展早期監(jiān)測(cè)的生物標(biāo)志物,為防治非小細(xì)胞肺癌提供幫助。
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(收稿日期:2018-08-09)