缺氧对人乳腺癌细胞系MCF-7生物学行为的影响  

The effects of hypoxia on the biological characteristics of human breast cancer cell line MCF-7

作　　者：赵婷婷[1] 邢鹏[1] 金锋[1] 李继光[2] 

机构地区：[1]中国医科大学附属第一医院乳腺外科,辽宁沈阳110001 [2]中国医科大学肿瘤研究所,辽宁沈阳110001

出　　处：《现代肿瘤医学》2014年第5期1015-1019,共5页Journal of Modern Oncology

摘　　要：目的:研究缺氧对人乳腺癌细胞系MCF-7的生物学行为,增殖、侵袭能力的影响。通过影响缺氧诱导因子1α(hypoxic-inducible factor 1α,HIF-1α)的表达,检验其在这一过程中的作用,并进一步探讨作用机制。方法:慢病毒感染细胞,干扰乳腺癌细胞系MCF-7中HIF-1α的表达,得到HIF-1α表达正常的乳腺癌细胞系MCF-7-NC和HIF-1α表达受干扰的细胞系MCF-7-HIF△。分别低氧培养及化学药物诱导缺氧,用MTT法检测2组细胞系在缺氧和正常培养条件下的增殖能力,ANOVA检验检测其增殖能力的区别。Transwell实验检验缺氧和正常培养条件下人乳腺癌细胞的侵袭能力,计数通过Transwell小室的细胞,ANOVA检验检测细胞侵袭能力的区别。共聚焦免疫荧光法检测缺氧后2组乳腺癌细胞的上皮和间质标记物的表达,检验细胞系是否发生了上皮-间质转化(epithelial-mesenchymal transition,EMT)。结果:MTT实验结果:正常表达HIF-1α的MCF-7-NC的增殖能力在缺氧后与正常培养条件下相比明显减弱,其差别具有统计学意义(P<0.05)。HIF-1α表达受干扰的MCF-7-HIF△的增殖能力在缺氧和正常培养条件下相比无显著区别。Transwell实验检测细胞的侵袭能力,缺氧后正常表达HIF-1α的MCF-7-NC的侵袭能力较正常培养的细胞明显增强,其差别具有统计学意义(P<0.05)。HIF-1α表达受干扰的MCF-7-HIF△侵袭能力在缺氧和正常培养下无显著区别。缺氧后,正常表达HIF-1α的细胞系MCF-7-NC发生上皮-间质转化(epithelial-mesenchymal transition,EMT)。共聚焦荧光染色显示:在诱导缺氧后,正常表达HIF-1α的MCF-7-NC细胞极性发生变化,呈长梭形改变,上皮标志物人广谱角蛋白(pan-cytokeratin,P-CK)表达下调,间质标志物人α平滑肌动蛋白(α-SMA)表达上调;而HIF-1α表达受干扰的MCF-7-HIF△无明显EMT变化。结论:在体外条件下,缺氧可以引起乳腺癌细胞系MCF-7增殖能力减弱、侵袭能力增强,可以诱导乳�Objective：To study the effects of hyPoxia on the biological characteristics of breast cancer cell line MCF - 7. With the interference of exPression of eIF - 1α,we tested the imPortant role of eIF - 1α（hyPoxic - induc-ible 1α）in this Process,moreover,to investigate the mechanism. Methods：Lentivirus infections were carried on to obtain the MCF - 7 - NC that exPression of eIF - 1α is normal,and MCF - 7 - eIF△ that exPression of eIF - 1α was interfered. Breast cancer cell lines were cultured under low O2 saturation（ ﹤ 1% O2 ）or adding chemical reagent des-ferrioxamine（DFO）to induce hyPoxia. MTT,Transwell assays were used to detect the effect of eIF - 1α exPression on the Proliferation and invasion of breast cancer cells. Confocal immunofluorescence was used to detect EMT of breast cancer cells. Results：MTT assays demonstrated the Proliferation of MCF - 7 - NC decreased visibly after hyPoxic cul-tivation（P ﹤ 0. 05）,while MCF - 7 - eIF△ showed no difference. Transwell assays illustrated the remarkable in-crease of invasion ability in MCF - 7 - NC after hyPoxic cultivation（P ﹤ 0. 05）,but hyPoxia seemed to have no influ-ence on the invasion ability of MCF - 7 - eIF△. After induced hyPoxia condition,EMT occurred in MCF - 7 - NC,the ePithelial markers down - regulated while the mesenchymal markers uP - regulated. In MCF - 7 - eIF△,EMT did not take Place. Confocal immunofluorescence confirmed the Phenomenon. Conclusion：eyPoxia affected breast cancer cell line MCF - 7,leads to the decrease of cell Proliferation and increase of invasion,as well as the EMT. When the ex-Pression of eIF - 1α was interfered,the effects of hyPoxia on breast cancer cells disaPPeared.

关 键 词：缺氧 HIF-1Α 乳腺癌 上皮-间质转化 

分 类 号：R737.9[医药卫生—肿瘤]