Kao C Y, Nomata K, Oakley C S, Welsch C W, Chang C C
Department of Pediatrics/Human Development, College of Human Medicine, Michigan State University, East Lansing 48824.
Carcinogenesis. 1995 Mar;16(3):531-8. doi: 10.1093/carcin/16.3.531.
A culture method to grow two morphologically distinguishable normal human breast epithelial cell types derived from reduction mammoplasty has been developed. Type I cells were characterized by a more variable cell shape, smooth cell colony boundaries, the expression of epithelial membrane antigen (EMA) and keratin 18 and the non-expression of keratin 14 and alpha 6 integrin. In addition, the Type I cells were growth stimulated by fetal bovine serum (FBS) and were deficient in gap junctional intercellular communication (GJIC). In contrast, Type II cells were characterized by a uniform cell shape, expression of keratin 14 and alpha 6 integrin and the non-expression of EMA and keratin 18. In addition, Type II cells were growth inhibited by FBS and were proficient in GJIC. Type I cells can be induced by cholera toxin to change their morphology to a Type II cell morphology. Hence, Type I cells antigenically resemble luminal epithelial cells, while the Type II cells more closely resemble basal epithelial cells. Type I and Type II cells were transfected with SV40 DNA. Clones with extended lifespans were obtained from both Type I and Type II cells by SV40 transfection. Some (2/9) of the SV40-transfected Type I cell clones became immortal (> 100 cumulative population doubling level), whereas none (0/8) of the SV40-transfected Type II cell clones became immortal. The SV-40-transfected Type I and Type II cell-derived extended life clones and immortal cell lines phenotypically resembled their parental cells with respect to EMA, keratin 14 and keratin 18 expression and GJIC. Each (9/9) of the SV40 transfected Type I cell clones grew in soft agar; none (0/8) of the SV40-transfected Type II cell clones were capable of growing in soft agar. These results provide evidence that normal human breast epithelial cells, derived from reduction mammoplasty, can be separated into two morphologically and antigenically different cell types and that these two different cell types significantly differ in their response to an oncogenic (SV40) stimulus.
一种用于培养从缩乳术获得的两种形态上可区分的正常人类乳腺上皮细胞类型的培养方法已被开发出来。I型细胞的特征是细胞形状更具变化性、细胞集落边界光滑、表达上皮膜抗原（EMA）和角蛋白18，不表达角蛋白14和α6整合素。此外，I型细胞受到胎牛血清（FBS）刺激生长，且缺乏间隙连接细胞间通讯（GJIC）。相比之下，II型细胞的特征是细胞形状均匀、表达角蛋白14和α6整合素，不表达EMA和角蛋白18。此外，II型细胞受到FBS抑制生长，且具备GJIC功能。I型细胞可被霍乱毒素诱导改变形态成为II型细胞形态。因此，I型细胞在抗原性上类似于管腔上皮细胞，而II型细胞更类似于基底上皮细胞。I型和II型细胞用SV40 DNA进行转染。通过SV40转染从I型和II型细胞中均获得了寿命延长的克隆。一些（2/9）SV40转染的I型细胞克隆变成了永生化细胞（累积群体倍增水平>100），而SV40转染的II型细胞克隆中没有一个（0/8）变成永生化细胞。SV - 40转染的I型和II型细胞衍生的寿命延长克隆及永生化细胞系在EMA、角蛋白14和角蛋白18表达以及GJIC方面在表型上类似于它们的亲代细胞。SV40转染的I型细胞克隆每一个（9/9）都能在软琼脂中生长；SV40转染的II型细胞克隆没有一个（0/8）能够在软琼脂中生长。这些结果提供了证据，表明从缩乳术获得的正常人类乳腺上皮细胞可以被分离成两种形态和抗原性不同的细胞类型，并且这两种不同的细胞类型对致癌（SV40）刺激的反应存在显著差异。
