背景:在微胶囊微环境中肿瘤细胞表现出更接近在体肿瘤的特性。目的:考察微囊化培养对肿瘤细胞耐药性的影响。方法:将平面培养至对数生长期的HepG2细胞微囊化培养15d,再次包封于微胶囊内进行微囊化培养,如此反复3次。回收每次微囊化培养的细胞,并通过显微镜、流式细胞仪、CCK-8和Real Time PCR检测细胞形态、黏附能力、增殖能力、细胞周期、药物敏感性和耐药相关基因的变化。结果与结论:微囊化培养不同次数的细胞再次进行平面培养,其形态、黏附能力、增殖能力和细胞周期均无显著变化。经过微囊化培养后再次进行平面培养,肿瘤细胞的耐药性随着微囊化培养次数的增加而逐渐下降,且耐药性降低的主要原因是耐药相关基因表达下调。提示只有在微胶囊这一独特微环境中肿瘤细胞才能保持高的耐药性,一旦回归平面培养,这一特性便会消失,肿瘤细胞只有在微囊化环境中培养才能表现出类似在体的耐药性。
BACKGROUND:Tumor cells within the microenvironment created by microcapsules have similar the characteristics of tumors in vivo. OBJECTIVE:To investigate the influence of microencapsulated culture on drug resistance of tumor cells. METHODS:HepG2 cells obtained from monolayer culture in the exponential growth phase were microencapsulated cultured for 15 days. Again the cells were encapsulated in microcapsules for the microencapsulated culture. The procedure above was repeated for three times. The cells after microencapsulated culture were retrieved for further examination. The morphology, adhesion ability, proliferation ability, cell cycle, drug sensitivity and the expression of drug resistance-associated genes were detected through microscope, flow cytometry, Cell Counting Kit-8 assay and real-time PCR, respectively. RESULTS AND CONCLUSION:The HepG2 cells went through different times of microencapsulated culture showed no significant changes upon their morphology, adhesion ability, proliferation ability and cell cycle when they were grown in monolayer culture again. Compared with the cells without microencapsulated culture, the drugs resistance of the retrieved cells from microcapsules decreased along with the increasing of the microencapsulated culturing times. It was primarily due to the expression of drug resistance-associated genes declined. The results of this study suggested only cultured in the microcapsules, the tumor cells could maintain the higher drug resistance, and once back to the monolayer culture, the characteristics would disappear. Therefore, the tumor cells could acquire the in vivo-like drug resistance only when cultured in the microcapsules.