Moreover, compound K, a bacterial metabolite of ginsenoside Rb1, right inhibits colon cancer cell development in vivo. Techniques Materials HCT116 cells have been obtained from ATCC. Male A J and immuno deficient nu nu mice were obtained from Jackson Laboratories. Azoxymethane was obtained from Midwest Research, the NCI Chemical Carcinogen Refer ence Conventional Repository. The sodium salt of dextran sul fate was obtained from MP Biomedicals. Ginseng extract was obtained from Wisconsin Ginseng Board. Com pound K protopanaxa diol was purchased from ChromaDex. The molecular excess weight of compound K is 6,222. 87. Harlan Tek lad prepared AIN 76A and Western and management chow diet programs. Details on the eating plan were as described. RC DC protein assay was obtained from Bio Rad. Rabbit polyclonal anti Cox two anti bodies were obtained from Cayman Chemical compounds.
Polyclonal antibodies to pAKT were obtained from Cell Signaling. Antibo dies to pEGFR, pErbB2, pERK, Bax, Bcl2, c Jun and cyclin D1 were obtained selleckchem from Santa Cruz Biotechnology. Antibodies to phospho active EGFR realize human EGFR phos phorylated tyrosine 1092 and antibodies to phospho active ErbB2 understand human ErbB2 phosphory lated tyrosine 1248. Monoclonal antibodies to b actin had been purchased from Sigma Aldrich. Antibodies to p27Kip1 and p21Waf1 had been bought from BD Transduction Laboratories. PCNA anti bodies had been obtained from Biomeda Corp. TUNEL assay kit was obtained from Millipore. Solutions Ginseng extraction and chemical analysis American ginseng root powder was obtained from Wis consin Ginseng Board.
5 hundred grams of ginseng was extracted in ten vol 70% ethanol underneath reflux for two h in the water bath kept at 90 95 C. The filtrate was collected plus the selleckchem Sorafenib extraction pro cedure repeated within the residue. The residue was dis carded just after the 2nd extraction. The filtrates had been combined and passed via filter paper. Filtrate was evaporated below vacuum. The extract was dissolved in 500 ml water and extracted 4 occasions in water saturated n butanol. The n butanol phase was evaporated under vacuum, and lyophilized. All solvents made use of for extraction were US pharmacopoeia pur ity. The ratio of commencing materials to ultimate ginseng extract was roughly 10,1. The ginsenosides within the extract have been characterized by UF HPLC as described. The ginsenoside concen trations have been Rg1, 14. 0, Re, 197. 9, Rb1, 341. eight, Rc, 34. two, Rb2, 4.
6, Rb3, six. 8, Rd, 65. 0, Rg3, 0. 6 as reported. Tumor Induction Utilization of animals for these scientific studies was approved below the guidelines of your Institutional Animal Care and Use Committee at University of Chicago, which complies using the pointers outlined through the National Institutes of Health and fitness. A J male mice weighing 20 22 grams were acclimated for 2 wks on AIN 76A chow. We followed a modified protocol to induce colitis asso ciated colon cancer.