Wagner H E, Joyce A D, Beatrice K, Summerhayes I C
New England Deaconess Hospital, Laboratory of Cancer Biology, Boston, Massachusetts 02115.
Oncogene. 1990 Apr;5(4):557-63.
To determine the in vivo phenotype elicited in bladder epithelium following the expression of a ras oncogene we have introduced the HaSV ras transforming gene into transplants of normal urothelium. Stripped adult bladder mucosa incubated with HaSV, in the presence or absence of helper virus, was transplanted beneath the renal capsule of syngeneic animals and maintained for periods up to three months. Control implants, exposed to helper virus alone, formed heterotopic bladders lined by urothelium displaying focal areas of full differentiation when associated with underlying submucosal elements. Exposure of urothelium to HaSV resulted in increased proliferative potential of mucosal and submucosal elements with the appearance of a normal differentiated heterotopic bladder 10 days post-implantation. Implants left for 28 days presented a range of hyperplastic lesions (mild-severe) characterized from histological evaluation and antibody markers recognizing basal (D66) and superficial cell populations (H10) in normal bladder mucosa. Staining of mild hyperplastic lesions revealed an increased basal cell compartment and a loss of fully differentiated cells lining the lumen of the implanted bladder. In severe hyperplasia no superficial cells were observed but mucosal elements stained throughout with antibody D66. This phenotype was accompanied by an irregular laminin staining pattern associated with a disorganized basement membrane and increased blood vasculature. Similar experiments conducted with HaSV and helper virus resulted in the generation of mesenchymal lesions at 28 days with little surviving urothelium. The H-ras oncogenic protein can induce hyperplastic lesions in normal urothelium which were characteristic of preneoplastic changes identified in bladder carcinogenesis.
为了确定在膀胱上皮中表达ras癌基因后所引发的体内表型，我们已将HaSV ras转化基因导入正常尿路上皮移植体中。将去除上皮的成年膀胱黏膜与HaSV一起孵育，无论有无辅助病毒，然后移植到同基因动物的肾被膜下，并维持长达三个月。仅暴露于辅助病毒的对照植入物形成了异位膀胱，当与下面的黏膜下层成分相关时，尿路上皮衬里显示出局部完全分化的区域。尿路上皮暴露于HaSV导致黏膜和黏膜下层成分的增殖潜能增加，植入后10天出现正常分化的异位膀胱。植入28天的植入物呈现出一系列增生性病变（轻度至重度），这些病变通过组织学评估以及识别正常膀胱黏膜中基底（D66）和表层细胞群（H10）的抗体标记物来表征。轻度增生性病变的染色显示基底细胞区增加，植入膀胱腔内衬里的完全分化细胞减少。在严重增生中，未观察到表层细胞，但黏膜成分用抗体D66全程染色。这种表型伴随着与紊乱的基底膜相关的不规则层粘连蛋白染色模式以及增加的血管。用HaSV和辅助病毒进行的类似实验在28天时导致间充质病变的产生，存活的尿路上皮很少。H-ras致癌蛋白可在正常尿路上皮中诱导增生性病变，这些病变是膀胱癌发生过程中所确定的癌前变化的特征。
