High SEPT9_v1 expression in human breast cancer cells is associated with oncogenic phenotypes.
Altered expression of the human septin gene, SEPT9, and its murine homologue, Sept9, has been implicated in neoplasia. However, their role(s) in oncogenesis remains poorly understood. We found amplification of SEPT9 in 67% of breast cancer cells (BCC) when compared with immortalized human mammary epithelial cells (IHMEC) as well as high levels of SEPT9 expression in the majority (61%) of the BCCs studied, unlike IHMECs. Expression profiling of variant SEPT9 transcripts and translated products revealed that high expression of the variant, SEPT9_v1, in contrast to other variants, was widespread in BCCs (55% of the BCCs) but not in IHMECs. High expression of SEPT9_v1 was also observed in primary breast cancer samples by immunohistochemical studies. We subsequently examined the phenotypic consequences of SEPT9_v1 expression in human breast cells. Retroviral expression of SEPT9_v1 in IHMEC cell culture models showed that SEPT9_v1 accelerated growth kinetics, stimulated cell motility, promoted invasion in Matrigel Transwell assays, increased genomic instability with the development of aneuploidy, and stimulated morphologic changes. Significant cytokinesis defects and disruption of tubulin microfilaments were also observed by immunofluorescence when SEPT9_v1 was ectopically expressed in IHMECs. Furthermore, SEPT9_v1 markedly enhanced neoplastic transformation in Hs578T cells, a BCC with no endogenous expression of the SEPT9_v1 isoform. Small interfering RNA-mediated and short hairpin RNA-mediated inhibition of SEPT9_v1 expression in two BCCs with high levels of endogenous SEPT9_v1 expression inhibited neoplastic growth properties of the cells. Taken together, our findings suggest that increased SEPT9_v1 expression contributes to the malignant pathogenesis of some breast tumors.
Gonzalez
Maria E
ME
Departments of Internal Medicine, University of Michigan, Ann Arbor, Michigan 48109, USA.
Peterson
Esther A
EA
Privette
Lisa M
LM
Loffreda-Wren
Janice L
JL
Kalikin
Linda M
LM
Petty
Elizabeth M
EM
D000782:Aneuploidy; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000096:biosynthesis* / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D005784:Gene Amplification; D006801:Humans; D008938:Mitosis / Q000502:physiology; D010641:Phenotype; D020033:Protein Isoforms; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D058112:Septins; D014404:Tubulin / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism
D020033:Protein Isoforms; D012333:RNA, Messenger; D014404:Tubulin; D014746:Vimentin; D020558:GTP Phosphohydrolases; C474315:SEPTIN9 protein, human; D058112:Septins