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目的:采用先进的荧光实时显像技术对中药土贝母植物块茎提取物抗乳腺癌作用进行评价。方法:MDA-MB-231人乳腺癌细胞被设计成为细胞质红色荧光蛋白(RFP)基因表达而细胞核组蛋白H2B绿色荧光蛋白(GFP)基因表达的双色荧光细胞,并分别在2D平面塑料平皿及3D立体明胶海绵块两种不同的培养环境中培养,经土贝母鲜品二氯甲烷提取物(DEFT)干预后,荧光共聚焦显微镜下观察细胞形态及核碎片形成的情况;DNA电泳方法进一步检测细胞凋亡。结果:土贝母鲜品二氯甲烷提取物作用24h后荧光显微镜下即观察到MDA-MB-231细胞出现凋亡的形态学改变;DNA电泳在药物干预48h后检测到凋亡的DNA碎片梯带,72h后更加明显;3D明胶海绵培养的MDA-MB-231细胞经实验药物作用后其形态学改变的效果不如2D环境下明显。结论:土贝母鲜品二氯甲烷提取成分具有抗乳腺癌治疗的潜能。
OBJECTIVE: To evaluate the anti-breast cancer effect of tuberous extract of Fritillaria cirrhosae by advanced fluorescent real-time imaging technology. METHODS: MDA-MB-231 human breast cancer cells were designed as two-color fluorescent cells expressing the cytoplasmic red fluorescent protein (RFP) gene and the nuclear histone H2B green fluorescent protein (GFP) gene and were respectively expressed in 2D flat plastic plates and 3D Three-dimensional gelatin sponge blocks were cultured in two different culture conditions. After the intervention of DEFT, the cell morphology and the formation of nuclear debris were observed under fluorescence confocal microscope. The DNA electrophoresis method was used to further detect Apoptosis. Results: Morphological changes of apoptotic MDA-MB-231 cells were observed under fluorescence microscope 24h after exposure to dichloromethane extract. DNA fragmentation was detected in DNA electrophoresis 48h after drug intervention The results showed that the morphological changes of MDA-MB-231 cells cultured in 3D gelatin sponge were not as obvious as those in 2D environment. Conclusion: The fresh extract of Tissue, Dichloromethane, has the potential of anti-breast cancer treatment.