Department of Dermatology and Harvard Skin Disease Research Center, Brigham and Women's Hospital and Harvard Medical School, 77 Ave. Louis Pasteur, Boston, MA 02115, USA.
In Vitro Cell Dev Biol Anim. 2010 Dec;46(10):841-55. doi: 10.1007/s11626-010-9353-8. Epub 2010 Nov 2.
Keratinocytes migrating from a wound edge or initiating malignant invasion greatly increase their expression of the basement membrane protein Laminin-322 (Lam332). In culture, keratinocytes initiate sustained directional hypermotility when plated onto an incompletely processed form of Lam332 (Lam332') or when treated with transforming growth factor beta (TGF-β), an inducer of Lam332 expression. The development and tissue architecture of stratified squamous and prostate epithelia are very different, yet the basal cells of both express p63, α6β4 integrin, and Lam332. Keratinocytes and prostate epithelial cells grow well in nutritionally optimized culture media with pituitary extract and certain mitogens. We report that prostate epithelial cells display hypermotility responses indistinguishable from those of keratinocytes. Several culture medium variables attenuated TGF-β-induced hypermotility, including Ca(++), serum, and some pituitary extract preparations, without impairing growth, TGF-β growth inhibition, or hypermotility on Lam322'. Distinct from its role as a mitogen, EGF proved to be a required cofactor for TGF-β-induced hypermotility and could not be replaced by HGF or KGF. Prostate epithelial cells have a short replicative lifespan, restricted both by p16(INK4A) and telomere-related mechanisms. We immortalized the normal prostate epithelial cell line HPrE-1 by transduction to express bmi1 and TERT. Prostate epithelial cells lose expression of p63, β4 integrin, and Lam332 when they transform to invasive carcinoma. In contrast, HPrE-1/bmi1/TERT cells retained expression of these proteins and normal TGF-β signaling and hypermotility for >100 doublings. Thus, keratinocytes and prostate epithelial cells possess common hypermotility and senescence mechanisms and immortalized prostate cell lines can be engineered using defined methods to yield cells retaining normal properties.
从伤口边缘迁移或开始恶性入侵的角质形成细胞大大增加了它们对基底膜蛋白层粘连蛋白-322（Lam332）的表达。在培养中，当角质形成细胞接种到不完全加工形式的 Lam332（Lam332'）或用转化生长因子-β（TGF-β）处理时，会引发持续的定向过度运动，TGF-β是 Lam332 表达的诱导剂。分层鳞状上皮和前列腺上皮的发育和组织结构非常不同，但两者的基底细胞都表达 p63、α6β4 整合素和 Lam332。角质形成细胞和前列腺上皮细胞在营养优化的培养基中生长良好，其中含有垂体提取物和某些有丝分裂原。我们报告说，前列腺上皮细胞表现出与角质形成细胞相似的过度运动反应。几种培养基变量减弱了 TGF-β诱导的过度运动，包括 Ca（++）、血清和某些垂体提取物制剂，但不会损害生长、TGF-β生长抑制或在 Lam322'上的过度运动。与作为有丝分裂原的作用不同，EGF 被证明是 TGF-β诱导的过度运动所必需的辅助因子，不能被 HGF 或 KGF 取代。前列腺上皮细胞的复制寿命很短，受 p16(INK4A)和端粒相关机制的限制。我们通过转导使正常前列腺上皮细胞系 HPrE-1 表达 bmi1 和 TERT 来实现永生化。前列腺上皮细胞在转化为侵袭性癌时会丧失 p63、β4 整合素和 Lam332 的表达。相比之下，HPrE-1/bmi1/TERT 细胞保留了这些蛋白质的表达以及正常的 TGF-β信号和过度运动能力，超过 100 次倍增。因此，角质形成细胞和前列腺上皮细胞具有共同的过度运动和衰老机制，并且可以使用定义的方法工程化永生化前列腺细胞系，以产生保留正常特性的细胞。
