Selection of carcinogen-altered rat tracheal epithelial cells preexposed to 7,12-dimethylbenz[a]anthracene by their loss of a need for pyruvate to survive in culture.
Selection of carcinogen-altered rat tracheal epithelial cells preexposed to 7,12-dimethylbenz[a]anthracene by their loss of a need for pyruvate to survive in culture.
复制标题
通过在培养物中不再需要丙酮酸来选择预先暴露于 7,12-二甲基苯并[a]蒽的致癌物改变的大鼠气管上皮细胞。
DOI：
10.1093/carcin/5.6.789
复制
发表时间：
1984
期刊：
影响因子：
4.7
通讯作者：
Martin,DH
中科院分区：
文献类型：
--
作者：
Marchok,AC;Huang,SF;Martin,DH
正常的大鼠气管上皮细胞需要外源性丙酮酸才能在培养中存活，而致癌物改变的细胞则不需要。我们研究的目的是：(I)利用致癌物质改变的细胞对丙酮酸的需求的丧失作为筛选和定量暴露于200μg 7，12-二甲基苯[a]-菲(DMBA)-蜂蜡球不同时间长度的气管中诱导的改变的细胞群数量的一种方式，(Ii)确定这些数量在暴露后以增加的时间间隔的稳定性，(Iii)寻找细胞群体中可能与暴露条件有关的独特的生长特性。将气管植入物暴露于200μg DMBA中2周、4周或连续暴露。在暴露2、6和9个月后，将气管切成碎片，并在富含威茅斯的培养基中从突起中建立原代培养物。2周后，从培养液中去除丙酮酸和胰岛素，持续2周，杀死正常细胞，离开改变的细胞群，即所选细胞群(SPC)。然后将丙酮酸和胰岛素返回到培养液中，以刺激SPC的快速扩张。DMBA暴露的时间长短对气管内诱导的SPC数量有显著影响。暴露2周后2个月的SPC为1.8个/根，此时4周和连续暴露的气管分别为5.0个和7.2个/根。SPC的数量在暴露后并没有随着时间的延长而增加，说明最大起始位点数是固定的。在2~9个月期间，2周暴露组可传代SPC的数量从67%增加到88%，而在较长时间的DMBA暴露后，93~100%的SPC是可传代培养的。与所有其他实验条件相比，只有2周DMBA-2个月组的SPC具有较少的细胞存活数量和较慢的平均生长速度。
Normal rat tracheal epithelial cells require exogenous pyruvate to survive in culture while carcinogen-altered cells do not. The aims of our study were, (i) to utilize this loss of requirement for pyruvate by carcinogen-altered cells as a way of selecting out and quantitating the numbers of altered cell populations induced in tracheas exposed to 200 μg 7,12-dimethylbenz[a]-anthracene (DMBA)-beeswax pellets for different lengths of time, (ii) to determine the constancy of these numbers at increasing time intervals after the exposure, (iii) to look for distinctive growth properties in cell populations that may be related to the exposure conditions. Tracheal implants were exposed for 2 weeks, 4 weeks or continuously to the 200 μg DMBA. At 2,6 and 9 months after the start of exposure, the tracheas were cut into pieces and primary cultures established from outgrowths in an enriched Waymouth's medium. After 2 weeks, pyruvate and insulin were removed from the medium for 2 weeks to kill the normal cells and leave the altered, i.e., the selected cell populations (SPC). The pyruvate and insulin were then returned to the medium to stimulate rapid expansion of the SPC. The length of DMBA exposure had a marked effect on the number of SPC induced in the tracheas. There were 1.8 SPC/trachea 2 months after the 2 weeks exposure, while 5.0 and 7.2 SPC were obtained from the 4 weeks and continuously exposed tracheas, respectively, at this time. The numbers of SPC did not increase with time after exposure indicating that the maximal number of initiation sites was fixed. Between 2 and 9 months the number of subculturable SPC increased from 67 to 88% in the 2 week exposed group, while 93–100% of the SPC were subculturable after the longer DMBA exposures. Only the SPC from the 2 week DMBA - 2 month group were distinguished by smaller numbers of cells surviving the selection medium and by a slower average growth rate compared to all the other experimental conditions.