Disease relevance of Col4a2

• The recombinant mouse canstatin efficiently expressed in E. coli M15 after IPTG induction was monitored by SDS-PAGE and by Western blotting with an anti-hexahistidine tag antibody [1].
• Nine copies of the hypoxia-response element were ligated upstream from the canstatin gene near the cytomegalovirus promoter [2].
• Angiogenesis is crucial for the growth and metastasis of solid tumors with sizes larger than a few cubic millimeter Canstatin, the non-collagenous 1 (NC1) domain of alpha2 chain of type IV collagen, was previously shown to inhibit proliferation of endothelial cells in vitro and suppress in vivo tumor growth [3].
• AIM: To examine the effect of canstatin, a newly discovered endogenous inhibitor of angiogenesis, in the treatment of pancreatic cancer in vivo [4].

High impact information on Col4a2

• Canstatin significantly inhibited human endothelial cell migration and murine endothelial cell tube formation [5].
• We demonstrate that apoptosis induced by canstatin was associated with a down-regulation of the anti-apoptotic protein, FLIP [5].
• Finally, we identified either a closely linked sequence related to Col4a2, or a recombination hot-spot between Col4a1 and Col4a2 that has been conserved in humans and mice [6].
• Isopropyl-1-thio-b-Dgalactopyran-oside (IPTG) was used to induce the expression of canstatin protein and affinity chromatography was used to purify the protein [4].
• In orthotopic pancreatic cancer models, the final tumor volume in groups treated with PBS, canstatin 5 mg/kg, 10 mg/kg were 355.21 +/- 39.54 mm(3), 112.73 +/- 10.47 mm(3), and 61.75 +/- 6.99 mm(3) respectively [4].

Anatomical context of Col4a2

• In addition, recombinant mouse canstatin potently inhibited endothelial cell proliferation with no inhibition on non-endothelial cells [1].
• The refolded mouse canstatin was tested on the chicken embryo chorioallantoic membranes (CAM), and a large number of newly formed blood vessels were significantly regressed [1].
• The expressed mouse canstatin, mainly as inclusion bodies, accounted for approximately 35% of the total bacterial proteins [1].
• Taken together, these findings demonstrate that the recombinant mouse canstatin effectively inhibited angiogenesis of the chicken embryo in a dose-dependent manner and specially suppressed in vitro the proliferation of human umbilical vein endothelial cells [1].

Regulatory relationships of Col4a2

• Transforming growth factor-beta (TGF-beta) treatment induced the expression of SOX9 and Col4a2, and a small interfering RNA against SOX9 reduced Col4a2 expression induced by TGF-beta treatment in mesangial cells [7].

Analytical, diagnostic and therapeutic context of Col4a2

• Canstatin protein expression was assessed by Western blot analysis in tumor tissues. pCMVCans and empty vector were used as controls in the in vivo assays [2].
• We believe that the hypoxia-inducible canstatin-expressing vector is a promising gene therapy tool for antiangiogenesis research [2].
• METHODS: The canstatin cDNA fragment was synthesized and amplified from the total RNA extracted from human placenta tissues by RT-PCR [4].
• Twenty-four nude mice with orthotopic xenograft tumor were randomly divided into 3 groups 10 d after the inoculation, and were treated with PBS 0.3 mL, or canstatin 5 mg/kg, or 10 mg/kg per day for 3 wk intraperitoneally [4].

References