Growth properties and tumorigenesis of MCF-7 cells transfected with isogenic mutants of rasH.
MCF-7 human breast cancer cells are estrogen dependent for maximal in vitro growth and for tumor formation in nude mice, thus providing a useful model system to study mammary tumorigenesis. A clone of MCF-7 cells transfected with the v-rasH oncogene has been shown to form tumors in the absence of estradiol [Kasid et al., 1985, Science (Wash. DC), 228:725-728]. To extend this observation to more clones of v-rasH-expressing MCF-7 cells and to examine the effects of rasH mutation, we transfected MCF-7 cells with a construct encoding the human c-rasH protooncogene protein product and with three isogenic constructs encoding proteins containing point mutations: arg-12, thr-59, and arg-12 + thr-59 (v-rasH). We isolated several cell lines which produced levels of c-rasH and v-rasH p21 at 30- to 50-fold the levels of controls. We also isolated several cell lines producing the various mutants p21s. All of the transfected cell lines were estrogen-responsive for cell growth. Transfected cells containing high levels of rasH p21 had correspondingly high levels of growth in an anchorage-independent growth assay. Tumorigenesis studies in nude mice, however, showed that some, but not all of the cell lines expressing v-rasH, formed tumors in the absence of estradiol. Tumor formation did not correlate with the level of rasH p21 expression in these cell lines. No tumor formation in the absence of estradiol was observed for cell lines expressing single-mutated or unmutated forms of rasH.
Division of Medical Oncology, Lombardi Cancer Center, Georgetown University Medical Center, Washington, DC 20007.
C L
CL
Sommers
A
A
Papageorge
G
G
Wilding
E P
EP
Gelmann
D000818:Animals; D001943:Breast Neoplasms / Q000473:pathology; D002455:Cell Division; D002460:Cell Line; D002478:Cells, Cultured; D005260:Female; D011905:Genes, ras; D006251:Harvey murine sarcoma virus / Q000235:genetics; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009154:Mutation; D009368:Neoplasm Transplantation; D010957:Plasmids; D011518:Proto-Oncogene Proteins / Q000032:analysis; D016283:Proto-Oncogene Proteins p21(ras); D014162:Transfection; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured / Q000166:cytology*
D011518:Proto-Oncogene Proteins; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)