John H. Frenster, Sharon R. Landrum, Marilyn A. Masek, and Lennard S. Wilson

Department of Medicine, Stanford University School of Medicine, Stanford, California 94305

Phone: +1 650 367 6483; Fax: +1 650 364 1773;  E-mail: frenster@euchromatin.net

Neoplastic cells are capable of undergoing cytoplasmic differentiation in-vitro (J. Natl. Cancer Inst. 54: 687 (1975). Nuclear changes in-vitro are less well characterized, but include hyper-segmentation of the cell nucleus, (Cancer 33: 716 (1974). In order to determine the ultrastructure and function of nuclear changes within neoplastic cells in-vivo, lymph nodes were fixed in glutaraldehyde within 30 seconds after removal from untreated patients with Hodgkin's Disease during original staging of their disease. 1 mm ³ blocks of fixed tissue were then reacted for 96 hours with Acridine Orange at 4^o C., washed to remove unbound probe, and digested with DNase I at 37^o C. for 30 min. as previously described (Cancer Res. 31: 1128 (1971). High-resolution electron microscopy was then used to localize and quantitate active DNA templates within each individual cell in its native location in the lymph node (Nature 248: 334 (1974). The maturation stages of the neoplastic Reed-Sternberg cells were separately identified by the ultrastructural criteria of previous studies (Natl. Cancer Inst. Monogr. 36: 239 (1973). Mononuclear Reed-Sternberg cells displayed higher DNA template activity/cell than did earlier Hodgkin's cells, while bi-nuclear Reed-Sternberg cells had still higher DNA template activity/cell. Multi-nuclear Reed-Sternberg cells displayed almost no DNA template activity/cell, but displayed the most extreme degrees of nuclear hyper-segmentation, blebbing of the nuclear membrane, and formation of nuclear pockets and projections. These quantitative ultrastructural data suggest that neoplastic cells in-vivo are capable of undergoing nuclear maturation sequences consisting of nuclear hyper-segmentation, blebbing, pocket and projection formation simultaneous with extreme degrees of inactivation of DNA template activity.

Supported in part by Research Grants from the National Cancer Institute (CA-10174) and the American Cancer Society (IC-45).

1. "Electron Microscopic Analysis of Lymph Node Cellular Activity in Hodgkin's Disease".
 

2. "Electron Microscopic Localization of Acridine Orange Binding to DNA within Human Leukemic Bone marrow Cells".

3. "Activity of DNA Templates During Cell Division and Cell Differentiation".

4. "Single-Cell Analysis of DNase I-Sensitive Sites during Neoplastic Cell Differentiation within Hodgkin's Disease Lymph Nodes".