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本文利用胞质杂交的细胞工程手段,研究人早幼粒白血病细胞突变株在添加小鼠网织红细胞胞质后的细胞恶性表型表达状态。实验结果表明,HGPRT~-人早幼粒白血病细胞突交株(HL-60-AR)与小鼠网织红细胞融合所形成的胞质杂交细胞,其恶性程度较亲本瘤细胞显著降低,它们在体外软琼脂培养基上不能形成集落,在裸鼠体内丧失致瘤能力,同时细胞增殖率降低,细胞分裂指数下降,DNA合成速率减慢。利用c-myc癌基因探针的分子杂交技术,未检测到胞质杂交细胞存在相当于c-myc基因特异的同源序列mRNA,提示杂交细胞癌基因表达严重受抑。对杂交细胞珠蛋白在翻译和转录水平上的表达分别进行探测,结果一致显示胞质杂交细胞中人珠蛋白基因受到激活。上述结果提示,小鼠网织红细胞中存在能调控人肿瘤细胞恶性表型和基因表达状态的胞质因子。
In this paper, the cytoplasmic hybrid cell engineering method was used to study the malignant phenotypic expression status of human promyelocytic leukemia cell mutants after addition of mouse reticulocyte cytoplasm. The experimental results showed that the cytoplasmic hybrid cells formed by the fusion of the HGPRT~-human promyelocytic leukemia cell line (HL-60-AR) with mouse reticulocytes have a significantly lower degree of malignancy than the parental tumor cells. Colonies could not be formed on the soft agar medium in vitro, and tumorigenicity was lost in nude mice. At the same time, the cell proliferation rate was decreased, the cell division index was decreased, and the DNA synthesis rate was slowed down. Using the c-myc oncogene probe molecular hybridization technique, no cytoplasmic hybrid cells were found to contain the c-myc gene-specific homologous sequence mRNA, suggesting that the expression of the oncogene in the hybrid cell was severely inhibited. The detection of the expression of the hybrid cell globin at the translational and transcriptional levels, respectively, revealed that the human globin gene was activated in the cytoplasmic hybrid cell. The above results suggest that there are cytoplasmic factors in mouse reticulocytes that can regulate the malignant phenotype and gene expression status of human tumor cells.