劉麗華il27轉(zhuǎn)染人食管鱗癌細(xì)胞抗腫瘤效應(yīng)及其機(jī)理研究

1、,IL-27轉(zhuǎn)染人食管鱗癌細(xì)胞抗腫瘤效應(yīng)及其機(jī)理研究,河北醫(yī)科大學(xué)第四醫(yī)院 科研中心河北省腫瘤研究所劉麗華,,,我國是食管癌的高發(fā)區(qū)，世界上約50%的食管癌發(fā)生在中國，到醫(yī)院就診的患者中95%為中晚期。盡管通過手術(shù)、化療和放療使食管癌的生存率得到很大提高，平均5年生存率仍為10%。近年來，腫瘤生物治療受到了人們的關(guān)注，并逐漸成為腫瘤的第四種治療方法。,引 言,IL-12 family members：IL-12, IL-

2、23 and IL-27 is primarily produced from activated antigen presenting cells (monocytes, macrophages and dendritic cells) whereas IL-35 is produced by regulatory T cells. These cytokines are all composed of two subunits,

3、a helical subunit (IL-12p35、IL-23p19、IL-27p28、IL-35p35) and a soluble cytokine receptor-like subunit (IL-12p40、IL-23p40、IL-27EBI3、 IL-35EBI3).,,,,,,,引言,本研究旨在探討IL-27基因轉(zhuǎn)染對(duì)人食管癌細(xì)胞在裸鼠體內(nèi)的抗腫瘤效應(yīng)及其對(duì)免疫功能的影響，探討其抗腫瘤作用機(jī)制。,目 的,細(xì)胞轉(zhuǎn)染

4、,,Eca109/IL-27 Eca109/LXSNEca109細(xì)胞,1×107個(gè)細(xì)胞/只,,,皮下接種,腹腔接種,,,成瘤性、生存期,,腫瘤體積,建立動(dòng)物模型,（mm3）＝長徑×短徑2×1/2,,IL-27對(duì)免疫調(diào)節(jié)作用研究,,Eca109/IL-27細(xì)胞的培養(yǎng)上清中檢測(cè)到IL-27，其含量為153.73±7.0pg/ml。而在Eca109/LXSN細(xì)胞和野生型Eca109細(xì)胞的培養(yǎng)上清中未

5、檢出IL-27。,蛋白水平鑒定轉(zhuǎn)染成功,,Fig.1 Expression of IL-27 EBI3 and p28 gene in different cells dected by RT-PCR. M: DNA marker; A:Eca109/IL-27 cells; B: Eca109/LXSN cells; C: Eca109 cells.,基因水平鑒定轉(zhuǎn)染成功,IL-27基因表達(dá),Fig.2 Tumor’s growth

6、 in nude mice inoculated subcutaneously with different tumor cells. * vs Eca109 and Eca109/LXSN groups respectively, P<0.01,IL-27對(duì)腫瘤生長的影響,生存期56±4day54±3day97±4day *,Fig.3 Infiltration of NK cells in

7、 tumor tissues from nude mice after inoculation with different cells. * vs Eca109 and Eca109/LXSN groups respectively, P<0.05,IL-27對(duì)移植瘤組織NK細(xì)胞浸潤數(shù)量的影響,(,*,Fig.5 Cytotoxic activity of spleen cells from nude mice after in

8、oculation different tumor cells. * vs Eca109 and Eca109/LXSN groups respectively, P<0.05,IL-27對(duì)荷瘤裸鼠脾細(xì)胞免疫功能的影響,*,Fig.4 IFN-g production from spleen cells after inoculation different tumor cells. * vs Eca109 and Eca109/

9、LXSN groups respectively, P<0.01,Fig.6 Gene expression in tumor tissues after inoculation different tumor cells detected by RT-PCR. A: Eca109; B: Eca109/LXSN; C: Eca109/IL-27,IL-27對(duì)移植瘤組織中細(xì)胞因子表達(dá)的影響,本研究結(jié)果表明，接種IL-27轉(zhuǎn)染細(xì)胞

10、組裸鼠移植瘤生長緩慢、裸鼠生存期延長，移植瘤組織中仍有IL-27的表達(dá)，表明IL-27在裸鼠體內(nèi)產(chǎn)生了抗腫瘤作用。IL-27受體高表達(dá)于NK細(xì)胞，因而我們進(jìn)一步檢測(cè)了移植瘤組織中NK細(xì)胞浸潤的數(shù)量。結(jié)果表明，接種IL-27轉(zhuǎn)染細(xì)胞組移植瘤組織間質(zhì)中NK細(xì)胞浸潤數(shù)量增高，效應(yīng)分子IFN-g產(chǎn)生增多，殺傷活性增強(qiáng)。說明IL-27不僅可以募集NK細(xì)胞而且能夠增強(qiáng)其生物學(xué)效應(yīng)。,討 論,,研究結(jié)果顯示接種Eca109/IL-27細(xì)胞

11、組移植瘤組織中T-bet和IL-12Rb2基因的表達(dá)水平增高。IL-27增強(qiáng)NK細(xì)胞產(chǎn)生IFN-g的分子基礎(chǔ)是IL-27與受體結(jié)合后活化STAT1，增加轉(zhuǎn)錄子T-bet表達(dá)，進(jìn)而作用于靶基因促進(jìn)IL-12Rb2和IFN-g等的轉(zhuǎn)錄。因此我們?cè)诜肿铀缴献C明IL-27能夠促進(jìn)NK細(xì)胞產(chǎn)生IFN-g 。,接種Eca109/IL-27細(xì)胞組移植瘤組織中Thl類細(xì)胞因子TNF-a、IL-1b和IL-6基因的表達(dá)亦升高。TNF-a在介導(dǎo)腫瘤細(xì)胞凋