Biochemical and Biophysical Research Communications
The candidate tumor suppressor CST6 alters the gene expression profile of human breast carcinoma cells: Down-regulation of the potent mitogenic, motogenic, and angiogenic factor autotaxin
Section snippets
Materials and methods
Cell culture and transfection. Human breast carcinoma MDA-MB-435S cells (passage #400) were purchased from ATCC and, before transfection, cloned by limiting dilution. A single clone, representative of the parental cell ‘line’ with regard to morphology and cystatin expression (parental clone), was stably transfected with a pTracer-CMV2-based CST6 expression vector (CST6-clones) or the ‘empty’ control vector (mock-clones) as reported previously [6]. Cells were seeded into 100-mm Petri dishes and
CST6 does not alter expression of other cystatins or cathepsins
To determine the molecular mechanism(s) by which CST6 suppresses the malignant properties of tumor cells in vitro [6], we compared global gene expression profiles in two mock- and two CST6-transfected human MDA-MB-435S cell clones. For this, we used Affymetrix Hu95A v.2 oligonucleotide microarrays, which interrogate about 12,625 transcript species. First, we determined whether ectopic expression of CST6 in MDA-MB-435S cells was compensated by increased expression of a lysosomal cysteine
Discussion
Normal epithelial cells undergo multiple genetic and epigenetic changes to acquire the highly proliferative, invasive, vasculogenic/angiogenic, immunosuppressive, and metastatic properties that are characteristic of malignant tumors [28]. These genetic changes lead in turn to global changes in gene expression [29], and more specifically, in overexpression of tumor-promoting genes and loss of expression of tumor-suppressing genes. Close to 100 genes have so far been identified as tumor-promoting
Acknowledgments
This study was in part supported by a Research Grant CA91785 (DK) and by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research (TC). JS received a Takeda Scholar-in-Training Award from the American Association for Cancer Research to present part of this work at the Special Conference on Cancer Research: ‘New Directions in Angiogenesis Research,’ Chicago, October 15–19, 2003. RS was partly supported by Research Grant CA36481 (Bonnie F. Sloane, Wayne
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Cystatins in cancer progression: More than just cathepsin inhibitors
2019, BiochimieCitation Excerpt :Next, cystatin E/M also acts via modulation of gene transcription as an additional mechanism to affect invasion [25,135]. In MDA-MB-435S cells cystatin E/M changed gene expression of ECM components, cytokines, kinases and phosphatases and several key transcription factors such as the signaling molecule autotaxin that is highly relevant for invasiveness and angiogenesis of breast carcinoma [194]. Overexpressed cystatin E/M in SCP2 cells suppressed breast carcinoma bone metastasis formation in vivo in nude mice models and reduced cancer aggressiveness.
Regulation of lysophosphatidate signaling by autotaxin and lipid phosphate phosphatases with respect to tumor progression, angiogenesis, metastasis and chemo-resistance
2011, BiochimieCitation Excerpt :ATX expression is strongly increased by the v-Jun oncogene [67] and α6β4 integrin, through the transcription factor, NFAT1, which binds to the ATX promotor at two distinct sites [61]. ATX is down-regulated by CST6, a tumor suppressor gene for breast cancer [68]. Induction of ATX expression by the Epstein–Barr virus promotes the growth and survival of Hodgkin lymphoma cells and specific knockdown of ATX decreases LPA levels and attenuates cell growth and viability [69].
Cystatin M expression is reduced in gastric carcinoma and is associated with promoter hypermethylation
2010, Biochemical and Biophysical Research CommunicationsCitation Excerpt :In gastric cancer, a growing number of genes have been identified as undergoing aberrant promoter hypermethylation, suggesting that promoter hypermethylation is an important mechanism involved in gastric cancer. Cystatin M is a member of a family of proteins that function as physiological inhibitors of lysosomal cysteine proteases, and control target proteases by forming high-affinity reversible complexes [6]. Loss of cystatin M expression might contribute to increased proteolysis of tissue architecture, facilitating the spread of cancer cells [4].
Rapid clearance of the circulating metastatic factor autotaxin by the scavenger receptors of liver sinusoidal endothelial cells
2009, Cancer LettersCitation Excerpt :Consistent with a role in angiogenesis, NPP2 promotes blood vessel formation in matrigel plug assays [5], while its knockout in mice is embryonic lethal due to a deficient angiogenesis [6,7]. The expression of NPP2 is induced by the tumor-stimulating proteins v-Jun [8], integrin α6β4 [9] and VEGF-A [10], and is downregulated by the tumor suppressor CST6 [11]. All cancer-related effects of NPP2 can be accounted for by its intrinsic lysophospholipase-D activity [12].
The biology of cystatin ME and its cognate target proteases
2009, Journal of Investigative DermatologyCitation Excerpt :This finding made the authors suggest that cystatin M/E may act as a tumor and metastasis suppressor through other mechanisms (than cathepsin inhibition), perhaps through its inhibitory effects on LGMN or even through modulation of gene transcription. Cystatin M/E expression in MDA-MB-435S cells significantly changed the expression of several genes involved in tumor growth, invasion, and angiogenesis, including downregulation of autotaxin, a signaling molecule that has been linked to breast cancer invasiveness (Song et al., 2006). Subsequently, the transfected MDA-MB-435S cells were injected into scid mice to test whether cystatin M/E expression might have in vivo tumor-suppressing functions (Zhang et al., 2004).
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Present address: Department of Pathology, Johns Hopkins Medical Institutions, Baltimore, MD 21231, USA.