人骨髓间充质干细胞诱导鼻咽癌上皮间质化的初步探讨

发布时间：2018-04-25 13:06

  本文选题：鼻咽癌 + 骨髓间充质干细胞　； 参考：《中华肿瘤防治杂志》2014年23期

【摘要】：目的初步探讨人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,hbMSCs)条件培养基(conditioned medium,CM)在人鼻咽癌CNE1及CNE2细胞上皮间质转化(epithelial mesenchymal transition,EMT)中的影响。方法取第3代hbMSCs融合达70%~80%时,换液培养24h后取其上清液(hbMSCs-CM);取对数生长期5×103CNE1及CNE2细胞,分别在普通培养液(对照组)和hbMSCs-CM的培养液(实验组)中培养。使用倒置相差显微镜下观察细胞形态学改变,蛋白质印迹法及q-PCR检测EMT相关蛋白及RNA变化。结果 hbMSCs-CM培养鼻咽癌细胞5d,CNE1和CNE2均观察到细胞间距离增宽,并且折光性强、多量单极或双极梭形状成纤维细胞样细胞,以CNE2细胞最为明显。q-PCR检测结果显示,与对照组相比,hbMSCs-CM培养组细胞上皮细胞标志基因mRNA的相对表达水平下调,差异有统计学意义,P值均0.001;间质细胞标志基因mRNA相对表达水平上调,差异有统计学意义,P值均0.001;EMT相关转录因子基因mRNA相对表达量上调,差异有统计学意义,P值均0.001。蛋白质印迹法检测结果显示,hbMSCs-CM诱导CNE1及CNE2细胞发生EMT并呈一定浓度依赖性。当hbMSCs-CM浓度≥70%,上皮细胞标志蛋白相对表达水平显著下降,差异有统计学意义,P0.05。当hbMSCs-CM浓度50%时,CNE1和CNE2细胞间质细胞标志及转录因子蛋白相对表达水平上调,差异有统计学意义,P0.05。结论 hbMSCs可改变鼻咽癌细胞形态,并下调细胞黏附蛋白及增加细胞间质蛋白水平,可能是参与鼻咽癌上皮间质转化过程的因素之一。
[Abstract]:Objective to investigate the effect of human bone marrow mesenchymal stem cells (hbMSCs) conditioned mediums CM) on epithelial mesenchymal transition of human nasopharyngeal carcinoma (CNE1) and CNE2 cells (CNE2 cells). Methods\\\; Methods when the third generation of hbMSCs fused to 70 ~ 80, the supernatant of HbMSCs-CMN was cultured for 24 hours, and the 5 脳 103CNE1 and CNE2 cells in logarithmic growth period were cultured in normal medium (control group) and hbMSCs-CM medium (experimental group), respectively. Morphologic changes were observed under inverted phase contrast microscope, EMT related proteins and RNA were detected by Western blot and q-PCR. Results for 5 days, hbMSCs-CM cultured nasopharyngeal carcinoma cell line CNE1 and CNE2 showed that the distance between cells was widened and the refractive index was strong. The results of multipolar or bipolar fusiform fibroblast-like cells were most obvious in CNE2 cells. Compared with the control group, the relative expression level of epithelial marker gene mRNA in hbMSCs-CM culture group was down-regulated, the difference was significant (P = 0.001), and the relative expression level of interstitial cell marker gene mRNA was up-regulated. The difference was statistically significant (P = 0.001). The relative expression of transcription factor gene mRNA was up-regulated, and the difference was statistically significant (P = 0.001). Western blot analysis showed that hbMSCs-CM induced EMT in CNE1 and CNE2 cells in a dose-dependent manner. When the concentration of hbMSCs-CM was 鈮，

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