In addition, compound K, a bacterial metabolite of ginsenoside Rb1, straight inhibits colon cancer cell development in vivo. Techniques Products HCT116 cells have been obtained from ATCC. Male A J and immuno deficient nu nu mice had been obtained from Jackson Laboratories. Azoxymethane was obtained from Midwest Study, the NCI Chemical Carcinogen Refer ence Typical 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 bought from ChromaDex. The molecular excess weight of compound K is six,222. 87. Harlan Tek lad ready AIN 76A and Western and manage chow diet programs. Details from the eating plan have been as described. RC DC protein assay was obtained from Bio Rad. Rabbit polyclonal anti Cox 2 anti bodies had been purchased from Cayman Chemical substances.
Polyclonal antibodies to pAKT were obtained from Cell Signaling. Antibo dies to pEGFR, pErbB2, pERK, Bax, Bcl2, c Jun and cyclin D1 had been obtained selleck inhibitor from Santa Cruz Biotechnology. Antibodies to phospho lively EGFR acknowledge human EGFR phos phorylated tyrosine 1092 and antibodies to phospho energetic ErbB2 understand human ErbB2 phosphory lated tyrosine 1248. Monoclonal antibodies to b actin had been bought from Sigma Aldrich. Antibodies to p27Kip1 and p21Waf1 were obtained from BD Transduction Laboratories. PCNA anti bodies have been obtained from Biomeda Corp. TUNEL assay kit was purchased 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 a water bath stored at 90 95 C. The filtrate was collected and also the selleckchem chk inhibitor extraction professional cedure repeated on the residue. The residue was dis carded immediately after the 2nd extraction. The filtrates were combined and passed via filter paper. Filtrate was evaporated underneath vacuum. The extract was dissolved in 500 ml water and extracted four instances in water saturated n butanol. The n butanol phase was evaporated beneath vacuum, and lyophilized. All solvents applied for extraction had been US pharmacopoeia pur ity. The ratio of beginning material to final ginseng extract was around 10,one. The ginsenosides from the extract have been characterized by UF HPLC as described. The ginsenoside concen trations had been Rg1, 14. 0, Re, 197. 9, Rb1, 341. 8, Rc, 34. two, Rb2, 4.
six, Rb3, six. eight, Rd, 65. 0, Rg3, 0. six as reported. Tumor Induction Utilization of animals for these research was accredited under the tips with the Institutional Animal Care and Use Committee at University of Chicago, which complies with all the recommendations outlined through the National Institutes of Wellness. A J male mice weighing twenty 22 grams had been acclimated for two wks on AIN 76A chow. We followed a modified protocol to induce colitis asso ciated colon cancer.