细胞因子诱导的杀伤细胞对卵巢癌耐药细胞SKOV3/CDDP的作用机制  

The functional mechanism of CIK cells on ovarian carcinoma cell lines SKOV3/CDDP

作　　者：张辉[1] 赵群[2] 左连富[3] 马洪俊[4] 李淑荣[4] 程建新[1] 贾金华[1] 康山 

机构地区：[1]河北医科大学第四医院妇产科,河北石家庄050011 [2]河北医科大学第四医院外科,河北石家庄050011 [3]河北医科大学第四医院肿瘤研究所,河北石家庄050011 [4]河北医科大学电镜室,河北石家庄050011

出　　处：《细胞与分子免疫学杂志》2007年第12期1167-1169,1171,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：河北省卫生厅科技攻关项目资助(2003/03086)

摘　　要：目的:探讨卵巢癌耐药细胞在癌变过程中免疫逃逸的分子机制以及细胞因子诱导的杀伤细胞(CIK)对其耐药性及免疫原性的影响。方法:采用电镜、流式细胞术(FCM)、MTT等检测方法,观察CIK细胞作用人卵巢癌耐药细胞系SKOV3/CDDP细胞超微结构、细胞周期、凋亡率、耐药相关基因MDR1和Topo-Ⅱβ以及hB7-1、hB7-2、MHCⅠb、HLA-DR抗原表达的变化;应用放射免疫(RIA)ELISA等方法检测CIK细胞作用于和荷人卵巢癌SKOV3/CDDP耐药细胞SCID小鼠腹腔移植模型后血清IL-2、TNF-α、IFN-γ、GM-CSF等细胞因子含量的改变。结果:电镜显示:CIK组可见较多的凋亡细胞,表现为细胞体积萎缩,膜发泡,细胞密度增加,线粒体浓缩染色质固缩于核膜周边;与生理盐水组相比,CIK组的G0/G1期细胞比例下降,S+G2/M期细胞比例升高,细胞周期阻滞于S+G2/M期,有统计学意义(P<0.05);在抑制细胞增殖的同时,尚诱导细胞凋亡,其细胞凋亡率为9.07%,二者比较,有统计学意义(P<0.05);与NS组相比,CIK组细胞MDR-1和Topo-Ⅱβ表达明显下降(P<0.05),提示CIK细胞可逆转人卵巢癌耐药细胞SKOV3/CDDP细胞耐药相关基因MDR1和Topo-Ⅱβ的表达;与NS组相比,CIK组细胞MHCⅠb、HLA-DR、hB7-1和hB7-2抗原的表达均明显升高(P<0.01)。提示CIK细胞可提高人卵巢癌耐药细胞株SK-OV3/CDDP的免疫原性;与NS组相比,CIK组小鼠血清中IL-2、TNF-α、INF-γ、GM-CSF含量均明显增加(P<0.01)。结论:CIK细胞在将卵巢癌耐药细胞的细胞周期阻滞于S+G2/M期的同时,尚能诱导其凋亡,降低耐药基因的表达,提高耐药细胞的免疫原性,分泌IL-2、TNF-α、INF-γ、GM-CSF等具有抗肿瘤活性的细胞因子发挥抗肿瘤作用。AIM： To investigate the functional mechanism of CIK cells or ovarian carcinoma cell lines SKOV3/ CDDP. METHODS： The changes of ultramicrostructure, cell cycle, apoptosis, expression of multidrug resistance-associated protein （MDR1, Topo-Ⅱ β） and other molecules （hB7-1, hB7-2, MHC Ⅰ b, HLA-DR） of SKOV3/CDDP cells treated with or without CIK cells were detected by electron microscope, MTr and FCM. The changes of cytokine （ IL-2, TNF-α, IFN-γ, GM-CSF） in sera of SCID mice bearing SKOV3/CDDP cells were detected by radioimmunit and ELISA. RESULTS： CIK cells could induce apoptosis of the SKOV3/CDDP cells by electronmicroscopic observations. The apoptosis rate in CIK group was 9.07%, and its cell cycle was arrested at S and G2/M phase （ P 〈 0.05 ）. Compared with NS Group, the co-expression of MDR-1 and Topo-Ⅱβ were decreased significantly in the CIK treated group （P〈0.05）, and the expression of MHCⅠb, HLA-DR, hB7-1 and hB7-2 antigen were increased significantly （ P 〈 0.01 ）. Compared with NS Group, the contents of IL-2, TNF-α, INF-γ, GM-CSF were increased significantly （ P 〈 0.01 ） in SCID mice of CIK group. CONCLUSION： CIK cells have several important biological effects on the ovarian carcinoma cell line SKOV3/CDDP, which may lay the foundation for further research on anti-tumor therapy.

关 键 词：卵巢癌 多药耐药 MHC抗原 共刺激分子 细胞因子诱导的杀伤细胞 

分 类 号：R392.11[医药卫生—免疫学]