摘要
目的:通过对体外培养的大鼠卵圆细胞进行AFP和c-ras、c-myc基因的动态测定,观察二者在卵圆细胞转化过程中的变化特点,探讨癌基因对细胞分化和转化的影响.方法:用化学致癌剂3’-Me-DAB诱发出SD大鼠肝卵圆细胞增生,采用Percoll密度梯度离心法分离之,并进行长期体外培养.在培养过程中,通过免疫荧光流式细胞仪租RNA-DNAslotblot杂交分别测定细胞AFP及癌基因的表达隋况.结果:AFP及c-ras、c-myc在卵圆细胞体外培养的不同时期呈现同步升高或降低:培养初期,卵圆细胞AFP及癌基因均呈高表达,而后下降;第20代时再次升高,之后维持在较低的表达水平.至第65代时,卵圆细胞不仅呈现出生长速度加快、群体倍增时间缩短、多倍体核型及软琼脂生长,而且癌基因及AFP的表达亦第3次升高.结论:癌基因及其产物不仅参与细胞的转化过程,亦可调节细胞的分化.
AIM: To study the effects of oncogenes on differentiation and transformation of oval cells by detecting and characterizing the expression of AFP and Ha-ras, c-myc genes of rat oval cells in vitro. METHODS: Proliferation of rat oval cells was induced by chemical carcinogen, 3'-Me-DAB. By using Percoll density gradient centrifugation method, oval cells were isolated, followed by continous cultivation in vitro. The expression of Ha-ras and c-myc genes and AFP in the oval cells from cultures was dynamically observed by RNA-DNA slot blot hybridization and flow cytometry. RESULTS: The expression of AFP and Ha-ras, c-myc genes in the cultured oval cells from different phases was synchronous: At the beginning of oval cell cultivation in vitro, both of AFP and oncogenes displayed a higher level expression and then declined. Up to 20th passage, the expression of AFP and oncogenes went up again and then kept a lower level. To 65th passage, the oval cells not only presented a growth rate increased, population doubling time shortened, adiploid chromosomes and growing on soft agar, but also the expression of oncogenes and AFP went up again. CONCLUSION: Oncogenes and their products participate not only in the regulation of cellular transformation, but also in the process of cell differentiation.
出处
《世界华人消化杂志》
CAS
2004年第2期344-346,共3页
World Chinese Journal of Digestology
基金
国家教委回国人员启动基金资助项目
No.2000479国家自然科学基金资助项目
No.30170473~~
