Elevated expression of SAG/ROC2/Rbx2/Hrt2 in human colon carcinomas: SAG does not induce neoplastic transformation, but antisense SAG transfection inhibits tumor cell growth.
Sensitive-to-apoptosis gene (SAG)/regulator of cullins (ROC)2/Rbx2/Hrt2 is a newly identified component of SCF E3 ubiquitin ligase that controls cell-cycle progression by promoting ubiquitination and degradation of cell-cycle inhibitors. We recently found that SAG protects cells from apoptosis induced by redox agents, promotes S-phase entry and cell growth under serum starvation, and is required for yeast growth. In the present study, we report that the SAG protein level was elevated in six of 10 human colon carcinoma tissues (60%) as compared with adjacent normal tissues from the same patient. SAG overexpression in preneoplastic cells in a JB6 tumor promotion-and-progression model did not induce neoplastic transformation, and SAG overexpression in NIH/3T3 cells did not induce transforming foci formation, suggesting that SAG is not a dominant oncogene. However, when DLD-1 human colon carcinoma cells were transfected with antisense SAG, monolayer growth was significantly inhibited, as shown by a decreased number of stable colonies in the plate after normalization with transfection efficiency. Stable clones that expressed antisense SAG showed a 50% decrease in their ability to form colonies when grown in soft agar versus clones that did not express antisense SAG. We found an inverse correlation in four of 10 tumors between the levels of SAG and p27, a cyclin-dependent kinase inhibitor. We concluded that SAG is not causally related to cellular transformation, but its overexpression may be important for the maintenance of tumor cell phenotype. Therefore, targeting SAG expression may have therapeutic value in cancer treatment. Mol. Carcinog. 30:62-70, 2001.
Department of Molecular Biology, Pfizer Global Research and Development, Ann Arbor Laboratories, Ann Arbor, Michigan 48105, USA.
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D000818:Animals; D002455:Cell Division / Q000235:genetics*; D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D003110:Colonic Neoplasms / Q000235:genetics* / Q000473:pathology; D004791:Enzyme Inhibitors / Q000378:metabolism; D006801:Humans; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D009124:Muscle Proteins; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D010641:Phenotype; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D014162:Transfection
D004791:Enzyme Inhibitors; D008840:Microfilament Proteins; D009124:Muscle Proteins; D016376:Oligonucleotides, Antisense; D011993:Recombinant Fusion Proteins; C058992:Tagln protein, mouse