Bodipy-C12在癌细胞过氧化物酶体中富集

目的:研究并比较Bodipy标记的月桂酸(Bodipy-C12)在癌细胞与正常细胞中的亚细胞定位。方法:在多种癌细胞和正常细胞的培养基中加入Bodipy-C12(1μg/m L),利用共聚焦显微镜活细胞定时间隔拍摄,结合不同细胞器的分子标记蛋白,观察Bodipy-C12五分钟内在不同细胞器中的定位。结果:在人肝癌细胞系HepG2细胞中,Bodpy-C12信号不仅仅存在于线粒体和脂滴,同时富集于过氧化物酶体中。我们分别采用Pex3-GFP、Pex14-GFP、GFP-Pex16、GFP-SKL和GFP-Pmp34等特异性定位的过氧化物酶体蛋白,确认Bodpy-C12信号富集于过氧化物酶体。此外,过氧化物酶体中Bodpy-C12信号的富集发生在更多的癌细胞系中,例如结肠癌细胞HCT116和乳腺癌细胞MCF7。不同的是,在正常细胞系如3T3-L1,NRK和COS7中,Bodipy-C12信号存在于线粒体和脂滴中,但未在过氧化物酶体中检测到。结论:Bodipy-C12信号存在于正常细胞和癌细胞的脂滴和线粒体中,且其在癌细胞过氧化物酶体中富集,而不存在于正常细胞的过氧化物酶体中,预示癌细胞中过氧化物酶体脂代谢的差异。

Bodipy-C12 can be Enriched in Peroxisomes of Cancer Cells

Objective:To investigate and compare the subcellular localization of Bodipy-labeled lauric acid(Bodipy-C12)in cancer cells and normal cells.Methods:Various cancer cells and normal cells were cultured in medium with Bodipy-C12(1μg/m L).We detected the signal of Bodipy-C12 in different organelles labeled by specific molecular maker proteins with using live-cell time-lapse confocal microscope in 5 minutes.Results:In a human hepatocellular carcinoma cell line HepG2,the signal of Bodpy-C12 was not only detected in lipid droplet and mitochondria,but it can also be enriched in peroxisomes.The co-localization of Bodpy-C12 signal with specific molecular protein markers,including Pex3-GFP,Pex14-GFP,GFP-Pex16,GFP-SKL and GFP-Pmp34,verified the existence of Bodpy-C12 in peroxisomes.Further,the enrichment of Bodpy-C12 in peroxisomes occured to more cancer cell lines such as colorectal cancer cell HCT116 and breast cancer cell MCF7.Differently,Bodpy-C12 signal was detected only in lipid droplet and mitochondria,but not in peroxisomes of normal cell lines such as 3 T3-L1,NRK,COS7.Conclusions:Bodipy-C12 signal existed in lipid droplet and mitochondria of normal cells and cancer cells,and it can be enriched in peroxisomes of cancer cells,but not in peroxisomes of normal cells,indicating a different lipid metabolism of the peroxisomes in cancer cells.