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目的通过增强型绿色荧光蛋白(pEGFP-1)的转化和导入ACC-M细胞株,筛选出ACC-M-GFP细胞,并在模拟的肺环境下,建立腺样囊性癌肺转移灶的动态观察模型。方法利用基因转化技术制备高纯度的pEGFP-1真核表达质粒。利用阳离子脂质体介导的基因转染技术培养ACC-M-GFP细胞。体内接种4周后建立模拟肺环境,并用激光共聚焦显微镜观察。结果从接种的第2周开始,随接种时间延长,荧光细胞集落的数量和荧光强度逐渐增加,到第4周时荧光表达强烈,多数成片分布。观察结果同时显示仅有极少数的转移细胞有能力形成集落。结论成功建立了腺样囊性癌肺转移灶的动态观察模型,这一模型为早期转移肿瘤细胞的捕获和分析提供了较好的平台。
OBJECTIVE: To screen ACC-M-GFP cells by transforming green fluorescent protein (pEGFP-1) and ACC-M cell line, and to establish the dynamic state of lung metastases of adenoid cystic carcinoma in a simulated lung environment Observe the model. Methods High purity pEGFP-1 eukaryotic expression plasmid was prepared by gene transformation technology. ACC-M-GFP cells were cultured using cationic liposome-mediated gene transfection. Four weeks after inoculation, the simulated lung environment was established and observed with laser confocal microscopy. Results From the second week of inoculation, the number and fluorescence intensity of fluorescent colony increased gradually with the prolongation of inoculation time. By the fourth week, the expression of fluorescence was strong and most of the fragments were distributed. Observations also show that only a very small number of metastatic cells are capable of forming colonies. Conclusion The dynamic observation model of lung metastasis of adenoid cystic carcinoma has been established successfully. This model provides a good platform for the early capture and analysis of metastatic tumor cells.