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Malignant Properties of Sublines Selected from a Human Bladder Cancer Cell Line That Contains an Activated c-Ha-ras Oncogene¹

Ontario Cancer Institute, Toronto, Ontario, Canada M4X 1K9

The human bladder cancer cell line MGH-U1 (also designated T-24 or EJ) contains an activated c-Ha-ras oncogene, which is amplified as compared to normal human fibroblasts. We have generated sublines from the MGH-U1 cell line: the MGH-U1/OCI subline was generated by dissociating spheroids formed from MGH-U1 cells; the U1-m/F₁ and OCI-m/F₁ were generated by in vivo passage of experimental lung metastases formed after i.v. injection of MGH-U1 and MGH-U1/OCI lines into immune-deprived mice; the U1/t subline was generated by in vivo passage of i.m. tumors formed from MGH-U1 cells. All sublines formed tumors in immune-deprived mice from smaller i.m. inocula than the parent line, and the U1-m/F₁ subline generated more spontaneous metastases in lungs. Lung colony forming efficiency after i.v. injections of cells into similar mice was also greater for the sublines than for the parent MGH-U1 cells. The U1-m/F₁ and OCI-m/F₁ were the most tumorigenic lines. Early passages of the MGH-U1/OCI subline showed the presence of double minute chromosomes, and amplification and increased expression of the c-Ha-ras oncogene as compared to the parental cell line. These changes were not present in later cultures of MGH-U1/OCI cells, and no consistent difference in the levels of gene amplification or expression between the parent line and the sublines was found. Thus the content and expression of the activated c-Ha-ras oncogene does not correlate with malignant properties of the sublines.

¹ Supported by research Grants CA29476 and CA29526 from the National Cancer Institute, NIH. Dr. Kovnat was a recipient of a National Cancer Institute of Canada Fellowship.

² To whom requests for reprints should be addressed, at Ontario Cancer Institute, 500 Sherbourne Street, Toronto, Ontario, Canada M4X 1K9.

Received 7/14/87. Revised 4/27/88. Accepted 6/ 6/88.

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