Kao C Y, Oakley C S, Welsch C W, Chang C C
Department of Pediatrics/Human Development, College of Human Medicine, Michigan State University, East Lansing 48824, USA.
In Vitro Cell Dev Biol Anim. 1997 Apr;33(4):282-8. doi: 10.1007/s11626-997-0048-8.
A chemically defined culture medium was developed to support the growth of two distinctly different types of normal human breast epithelial cells (HBEC) derived from reduction mammoplasty. Type I cells expressed luminal epithelial cell markers and were deficient in gap junctional intercellular communication (GJIC), whereas Type II cells expressed basal epithelial cell markers and were efficient in GJIC. In this study, we examined and compared the growth factor and hormone requirements of these two types of cells and a series of cell lines that were obtained by sequential transfection with SV40 DNA (extended lifespan, nontumorigenic), treatment with 5-bromodeoxyuridine (BrdU)/black light (immortal and weakly tumorigenic), and infection of a virus carrying the neu oncogene (highly tumorigenic). Growth of Type I cells was inhibited by withdrawing epidermal growth factor (EGF), hydrocortisone (HC), or insulin (INS) from the culture media, but was enhanced by fetal bovine serum (FBS) supplementation. Growth of Type II cells was inhibited by withdrawal of EGF, HC, or INS from the media, and was inhibited by FBS supplementation. Withdrawal of human transferrin (HT) or 17 beta-estradiol (E2) from the media did not alter the growth of Type I or Type II cells. SV40 transfected Type I cell lines still required EGF, HC, or INS for optimal growth. However, the highly tumorigenic cell line did not show a growth dependence on EGF, HC, or INS but did appear to require HT and 3,3',5-triiodo-D.L. thyronine (T3) for optimal growth. In addition, FBS stimulated the growth of these cell lines. Thus, this study shows that Type I HBEC are distinctly different from Type II HBEC in growth response to FBS and that neoplastically transformed Type I cells could become growth factor and hormone independent.
开发了一种化学成分明确的培养基，以支持从缩乳术获得的两种截然不同类型的正常人乳腺上皮细胞（HBEC）的生长。I型细胞表达腔上皮细胞标志物，细胞间间隙连接通讯（GJIC）功能缺陷，而II型细胞表达基底上皮细胞标志物，GJIC功能正常。在本研究中，我们检测并比较了这两种细胞以及一系列通过用SV40 DNA进行连续转染（延长寿命，非致瘤性）、用5-溴脱氧尿苷（BrdU）/黑光处理（永生化且弱致瘤性）和感染携带neu癌基因的病毒（高致瘤性）获得的细胞系对生长因子和激素的需求。从培养基中去除表皮生长因子（EGF）、氢化可的松（HC）或胰岛素（INS）会抑制I型细胞的生长，但补充胎牛血清（FBS）可促进其生长。从培养基中去除EGF、HC或INS会抑制II型细胞的生长，补充FBS也会抑制其生长。从培养基中去除人转铁蛋白（HT）或17β-雌二醇（E2）不会改变I型或II型细胞的生长。SV40转染的I型细胞系仍需要EGF、HC或INS以实现最佳生长。然而，高致瘤性细胞系对EGF、HC或INS没有生长依赖性，但似乎需要HT和3,3',5-三碘-D.L.甲状腺原氨酸（T3）以实现最佳生长。此外，FBS刺激这些细胞系的生长。因此，本研究表明，I型HBEC在对FBS的生长反应上与II型HBEC明显不同，并且肿瘤转化的I型细胞可能变得不依赖生长因子和激素。
