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目的:建立小鼠多药耐药白血病细胞系P388/VCR并研究其生物学特性。方法:小鼠淋巴细胞白血病细胞系P388,通过长期、单一、逐步提高浓度的方法建立多药耐药的白血病细胞系P388/VCR。采用悬浮细胞培养和半固体培养观察该细胞系的生长规律;流式细胞术分析细胞周期分布及柔红霉素在细胞内的积聚量;RT蛳PCR方法检测mdrla基因表达;四氮甲基唑蓝(MTT)法研究P388/VCR细胞系的耐药谱。结果:经过6个月单一、小剂量及间歇给药的方法诱导P388细胞成为多药耐药细胞系P388/VCR。与亲本细胞系P388比较,细胞生长曲线及倍增时间无明显差异,软琼脂培养集落形成率及大集落形成率差异不显著,细胞周期分析P388/VCR细胞S期比例降低。P388/VCR细胞除对原诱导药物具有耐药性外,对其他多种抗肿瘤药物如柔红霉素、高三尖杉酯碱、米托蒽醌、足叶乙苷等也具有交叉耐药性。流式细胞仪检测细胞内柔红霉素含量P388/VCR细胞平均荧光值为3.72,明显低于P388细胞的17.31。RT蛳PCR检测mdrla基因的表达结果为:P388细胞阴性,P388/VCR细胞阳性。结论:多药耐药细胞系P388/VCR具有mdrla基因表达阳性及对多种抗肿瘤药物耐药的特征。
Objective: To establish a mouse multidrug resistant leukemia cell line P388 / VCR and study its biological characteristics. METHODS: The murine lymphocytic leukemia cell line P388 was established by establishing a multidrug resistant leukemia cell line P388 / VCR by long-term, single and gradual increase in concentration. The growth of the cell line was observed by suspension cell culture and semi-solid culture. The cell cycle distribution and the amount of daunorubicin in cells were analyzed by flow cytometry. The mdrla gene expression was detected by RT 蛳 PCR. Blue (MTT) method to study drug resistance spectrum of P388 / VCR cell line. Results: P388 cells were induced to multidrug resistance cell line P388 / VCR after single, small dose and intermittent administration for 6 months. Compared with the parental cell line P388, there was no significant difference in cell growth curve and doubling time. There was no significant difference in colony formation rate and colony formation rate between soft agar culture and S phase in cell cycle analysis P388 / VCR cells. P388 / VCR cells in addition to the original drug-induced resistance, but also on a variety of other anti-tumor drugs such as daunorubicin, homoharringtonine, mitoxantrone, etodide also cross-resistant . The average fluorescence of P388 / VCR cells detected by flow cytometry was 3.72, which was significantly lower than that of P388 cells (17.31). RT 蛳 PCR detection of mdrla gene expression results: P388 cells were negative, P388 / VCR cells were positive. Conclusion: Multidrug-resistant cell line P388 / VCR has the characteristics of mdrla gene expression and resistance to a variety of anti-tumor drugs.