IL-8通过上调Bcl-2的表达和下调caspase-3的表达抑制MCF-7乳腺癌细胞凋亡  

IL-8 inhibits the apoptosis of MCF-7 human breast cancer cells by up-regulating Bcl-2 and down-regulating caspase-3

作　　者：庞雪利[1] 李矿发[1] 魏兰[1] 黄云秀[1] 苏敏[1] 王林[1] 曹红[1] 陈婷梅[1] 

机构地区：[1]重庆医科大学教育部临床检验诊断学重点实验室,重庆400016

出　　处：《细胞与分子免疫学杂志》2015年第3期307-311,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81272544)

摘　　要：目的探讨白细胞介素8(IL-8)对乳腺癌细胞MCF-7凋亡的影响及其机制。方法 Western blot法检测MCF-7细胞IL-8受体CXC趋化因子受体1(CXCR1)、CXCR2的表达;反转录PCR、Western blot法检测(0、20、40、80、160)ng/mL IL-8对MCF-7细胞Bcl-2、caspase-3表达的影响;CCK-8法检测(0、40、80)ng/mL IL-8对MCF-7细胞增殖的影响;相差显微镜下观察80ng/mL IL-8处理MCF-7后细胞形态的变化;Western blot法检测80ng/mL IL-8联合信号通路抑制剂10μmol/L PD980590、10μmol/L LY294002或50μmol/L AG490[分别为丝裂原活化蛋白激酶/细胞外调节蛋白激酶(MAPK/ERK)、磷酸肌醇-3激酶/蛋白激酶B(PI3K/AKT)、Janus激酶/信号转导子和转录激活子(JAK/STAT)信号通路抑制剂],共同处理MCF-7细胞后,细胞内Bcl-2蛋白表达的变化;Western blot法检测(0、20、40、80、160)ng/mL IL-8对MCF-7细胞磷酸化p-AKT表达的影响;流式细胞术、反转录PCR以及Western blot法分别检测80ng/mL IL-8联合10μmol/L LY294002共同处理MCF-7细胞后,细胞凋亡以及细胞内Bcl-2、caspase-3表达的变化。结果 IL-8受体CXCR1、CXCR2在MCF-7细胞中均有表达;在IL-8的作用下,MCF-7细胞Bcl-2表达升高,caspase-3表达下降,抗凋亡能力明显增强;IL-8能显著上调MCF-7细胞中p-AKT的表达;PI3K/AKT信号通路抑制剂LY294002能显著抑制IL-8抗MCF-7细胞凋亡的作用,且减少Bcl-2并增加caspase-3的表达。结论 IL-8可显著抑制MCF-7细胞的凋亡,其机制可能与IL-8激活PI3K/AKT信号通路而上调Bcl-2、下调caspase-3的表达有关。Objective To investigate the effect of interleukin-8 （IL-8） on the apoptosis of MCF-7 human breast cancer cells and the molecular mechanism. Methods The expressions of IL-8 receptors （CXCR1, CXCR2） in MCF-7 cells were detected by Western blotting. The effects of 0, 20, 40, 80, 160 ng/mL IL-8 on the expressions of apoptosis-related genes Bcl-2 and caspase-3 in MCF-7 cells were observed by reverse transcription PCR （RT-PCR） and Western blotting. Cell proliferation was determined by CCK-8 assay after 0, 40, 80 ng/mL IL-8 treatment. Phase contrast microscope was used to examine cell morphology of MCF-7 cells after 80 ng/mL IL-8 treatment. The effects of 80 ng/mL IL-8 combined with PD980590 （10 μmol/L）, LY294002 （10 μmol/L） or AG490 （50 μmol/L） [ mitogen-activated protein kinase/extracellular signal-regulated kinase （ MAPK/ERK）, Janus kinase/signal transducer and activator of transcription （JAK/STAT） signal pathway inhibitors, respectively], on the expression of Bcl-2 were detected by Western blotting. The effects of 0, 20, 40, 80, 160 ng/mL IL-8 on the expression of p-AKT in MCF-7 cells were observed by Western blotting. The effects of 80 ng/mL IL-8 combined with 10 ~mol/L LY294002 on the apoptosis of MCF-？ cells, the expressions of Bcl-2 and caspase-3 were determined by flow cytometry, RT-PCR and Western blotting, respectively. Results Both CXCR1 and CXCR2 were expressed in MCF-7 cells. IL-8 markedly up-regulated the anti-apoptotic gene Bcl-2, down-regulated the pro-apoptotic gene caspase-3, and significantly inhibited the apoptosis of MCF-7 cells. However, these effects were blocked by phosphoinositide 3-kinase/protein kinase B （PI3K/AKT）, signal pathway inhibitor LY294002. As predicted, IL-8 markedly increased the expression of p-AKT in MCF-7 cells. Conclusion IL-8 might significantly inhibit the apoptosis of MCF-7 cells. This effect may be achieved by up-regulating Bcl-2 and down-regulating caspase-3 via PI3K/AKT signal pathway.

关 键 词：IL-8 乳腺癌 MCF-7细胞 凋亡 

分 类 号：R737.9[医药卫生—肿瘤] Q279[医药卫生—临床医学]