全反式维甲酸改变人乳腺癌细胞外基质微环境的研究

The study of microenvironment modification of extracellular matrix in breast cancer by all-trans retinoic acid

目的:研究全反式维甲酸(ATRA)对乳腺癌细胞MCF-7增殖、分化诱导、细胞微环境改变及对乳脂球表皮生长因子8(MFGE8)转录表达的影响,为乳腺癌防治提供理论依据。方法:油红O染色检测ATRA对MCF-7细胞的分化诱导作用及对细胞微环境的影响;台盼蓝染色检测ATRA对MCF-7细胞的增殖抑制率;实时定量PCR检测ATRA对β-casein和对MFGE8转录表达的影响。结果:ATRA对MCF-7细胞有很强的分化诱导作用且呈剂量依赖性,高浓度ATRA诱导MCF-7细胞所形成的脂肪滴大于低浓度时的脂肪滴,脂肪滴主要集中于细胞膜外周及胞浆,改变了细胞微环境及形态结构;ATRA对细胞的增殖抑制率呈剂量依赖性。2μmol·L^(-1)ATRA处理细胞1~5 d,第4天β-casein转录表达增强最为显著,为7.26倍;第3天MFGE8转录表达增强最为显著,为6.74倍。结论:ATRA诱导MCF-7细胞分化,抑制细胞生长,改变其微环境,进而促使癌细胞逆转。

Objective： This study was designed to investigate the effects of ATRA on the proliferation,differentiation,microenvironment and MFGE8 transcription of breast cancer cells（ MCF-7）,which will provide theoretical basis for breast cancer therapy. Methods： Oil red O staining was used to estimate the differentiation induction and microenvironment modification of MCF-7 by ATRA. The inhibitory effect of ATRA on MCF-7 was determined by trpan blue exclusion. The effect of ATRA on -casein and MFGE8 transcription was demonstrated by real time PCR. Results： MCF-7 differentiation was obviously induced by ATRA in a dose-dependent manner.Lipid droplets induced by high concentrations of ATRA were much larger in size than that induced by low concentrations. Lipid droplets located in cytoplasm and outside of cell membranes modified the microenvironment and the cell structure. The inhibition rates of cell proliferation by different concentrations of ATRA were in a dosedependent manner. After MCF-7 treatment with 2 μmol · L^- 1ATRA for 1 to 5 days,β-casein transcription increased most by 7. 26-fold on the fifth day. In the meantime,MFGE8 transcription increased most by 6. 74-fold on the third day. Conclusion： ATRA exerts the effects on the proliferation,differentiation and microenvironment of MCF-7 cells,which can be further reprogrammed and reversed.