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Changes in Growth and Tumorigenicity following Reconstitution of Retinoblastoma Gene Function in Various Human Cancer Cell Types by Microcell Transfer of Chromosome 13¹

Center for Biotechnology, Baylor College of Medicine, The Woodlands, Texas 77381 [A. B., H-J. X., S-X. H., R. T., W. F. B.], and Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine, California 92717 [D. A., E. J. S.]

Functional loss of the retinoblastoma (RB) gene has been implicated in the initiation or progression of several human tumor types including cancer of the eye, bone, bladder, and prostate. To examine the consequence of adding one RB allele containing its normal regulatory elements back into representative examples of each of these cancer types, as well as to compare the results to those previously reported using various RB complementary DNA contructs, a neomycin resistant marked 13 chromosome was transferred by microcell fusion. Several attempts to obtain RB positive osteosarcoma cells failed. In addition, only one RB positive retinoblastoma clone was isolated. This clone contained many large cells, could not be maintained in long-term culture, and produced only RB negative tumors. Three RB positive bladder cancer cell clones were obtained, all of which grew slower in culture than their RB negative parental counterpart and did not form colonies in soft agar. Tumorigenicity was markedly suppressed in these clones. One clone yielded no tumors, and the other 2 clones produced only one small tumor each, both of which were RB negative. In contrast, the 2 RB positive prostate cancer cell clones isolated had no differences in their cell culture growth properties, including growth in soft agar compared to the parental cells. One of the clones was nontumorigenic, while the other clone produced 4 small tumors, all of which were RB positive. These results indicate that the transfer of one RB allele by microcell transfer produces different levels of growth inhibition as well as tumor suppression, depending on the cell type examined. In the case of prostate cancer, the function of the RB gene in tumor suppression appears to be independent from its growth regulatory function, since no growth inhibition in cell culture was noted in these cells, although tumor suppression was significant.

¹ This study was supported by NIH Grants EYO6195 and CA29401, a grant from the Retina Research Foundation, and The Council for Tobacco Research-United States of America.

² To whom requests for reprints should be addressed, at the Center for Biotechnology, Baylor College of Medicine, 4000 Research Forest Drive, The Woodlands, TX 77381.

Received 6/ 1/92. Accepted 9/11/92.