Modulation of cystatin C expression impairs the invasive and tumorigenic potential of human glioblastoma cells.
Increases in the abundance of cathepsin B transcript and protein with increased tumor grade and changes in subcellular localization and activity of this enzyme. We observed progressive reductions in levels of the protease inhibitor cystatin C, an inhibitor of cathepsin B with corresponding increases in the malignancy of glioma cell lines, implying an inverse correlation between cystatin C and tumor grade. To investigate the role of cystatin C in the invasion of brain tumor cells, we stably transfected SNB19 glioblastoma cells with either a 0.4-kb cDNA construct of human cystatin C in the sense orientation or an empty vector. Clones expressing sense-cystatin C cDNA had higher cystatin C mRNA and protein levels than did control cells. Sense-transfected cells were also markedly less invasive than control cells in a Matrigel invasion assay and in a coculture assay of SNB19 spheroids and fetal rat brain aggregates. Finally, the sense-transfected cells did not form tumors in nude mice upon intracerebral injection. These results strongly implicate cystatin C in the invasiveness of human glioblastoma cells and suggest that sense transcripts of cystatin C may prove useful in cancer therapy.
Konduri
Santhi D
SD
Division of Cancer Biology, Department of Biomedical and Therapeutic Sciences, University of Illinois, Peoria, Illinois 61656, USA.
Yanamandra
Niranjan
N
Siddique
Khawar
K
Joseph
Arun
A
Dinh
Dzung H
DH
Olivero
William C
WC
Gujrati
Meena
M
Kouraklis
Gregory
G
Swaroop
Amand
A
Kyritsis
Athanassios P
AP
Rao
Jasti S
JS
D000818:Animals; D001932:Brain Neoplasms / Q000235:genetics / Q000473:pathology*; D018920:Coculture Techniques; D055316:Cystatin C; D015891:Cystatins / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D005909:Glioblastoma / Q000235:genetics / Q000473:pathology*; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured
C525030:CST3 protein, human; C525031:Cst3 protein, mouse; C525032:Cst3 protein, rat; D055316:Cystatin C; D015891:Cystatins; D012333:RNA, Messenger