title: The aspartic protease napsin A suppresses tumor growth independent of its catalytic activity.
abstract: Members of the aspartic protease family have been implicated in cancer progression. The aspartic protease napsin A is expressed in type II cells of the lung, where it is involved in the processing of surfactant protein B (SP-B). Napsin A is also expressed in kidney, where its function is unknown. Here, we examined napsin A mRNA expression in human kidney tissues using in situ hybridization. Whereas strong napsin A mRNA expression was observed in kidney proximal tubules, expression was detected in only one of 29 renal cell carcinomas. This result is consistent with previous observations of loss of napsin A expression in high-grade lung adenocarcinomas. We re-expressed napsin A in the tumorigenic HEK293 kidney cell line and examined the phenotype of stably transfected cells. Napsin A-expressing HEK293 cells showed an altered phenotype characterized by formation of cyst-like structures in three-dimensional collagen cultures. Napsin A-expressing cells also showed reduced capacity for anchorage-independent growth and formed tumors in SCID mice with a lower efficiency and slower onset compared to vector-transfected control cells. Mutation of one of the aspartic acid residues in the napsin A catalytic site inactivated enzymatic activity, but did not influence the ability to suppress colony formation in soft agar and tumor formation. The mutation of the catalytic site did not affect processing, glycosylation or intracellular localization of napsin A. These data show that napsin A inhibits tumor growth of HEK293 cells by a mechanism independent of its catalytic activity.
authors.affiliation: Department of Oncology-Pathology, Cancer Center Karolinska, Karolinska Institute and Hospital, Stockholm, Sweden. takayuki@kuhp.kyoto-u.ac.jp
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ueno
authors.forename: Göran
authors.initials: G
authors.lastname: Elmberger
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Weaver
authors.forename: Masakazu
authors.initials: M
authors.lastname: Toi
authors.forename: Stig
authors.initials: S
authors.lastname: Linder
mesh_terms: D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000627:therapeutic use; D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000473:pathology; D002384:Catalysis; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002999:Clone Cells; D003094:Collagen / Q000737:chemistry; D018076:DNA, Complementary; D006639:Histidine / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000166:cytology; D008113:Liver Neoplasms / Q000201:enzymology* / Q000473:pathology; D051379:Mice; D016513:Mice, SCID; D009154:Mutation; D009369:Neoplasms / Q000188:drug therapy*; D012333:RNA, Messenger / Q000032:analysis; D013997:Time Factors; D014162:Transfection; D014183:Transplantation, Heterologous; D047368:Tumor Burden
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D006639:Histidine; D003094:Collagen; D016282:Aspartic Acid Endopeptidases; C498549:NAPSA protein, human