E MCF-7 and its derivative doxorubicin ABCB1 overexpressing MCF 7/adr, carcinoma Epidemo Of human KB cell line and its derivative, selected Hlt vincristine ABCB1 overexpressing KBv200, the S1-C London Online carcinoma and its derivative mitoxantrone selected Hlt ABCG2 overexpression S1 M1 80th HEK293/pcDNA3.1 ABCG2 were 482 R5, ABCG2 ABCG2 482 G2 and 482-T7 cells by selection with ZM-447439 331771-20-1 G418 after transfection with either empty pcDNA3.1 vector or HEK293 vector pcDNA3.1 full length Length contains Lt ABCG2 coding either set the arginine, glycine or threonine at the amino ureposition 482, respectively, and were in a medium containing 2 mg / ml G418. All cells were grown in culture medium free of drug for 2 weeks before the test. The cytotoxicity Tsassay MTT assay was used to cytotoxicity to translate t.
In detail, the cells in 96-well microtiter plates were cultured. To the toxicity of t to be determined by lapatinib, lapatinib different concentrations were diluted with medium to where the wells. To investigate the effect of lapatinib on the Chemosensitivit t of cancer cells, lapatinib the medium with various concentrations of doxorubicin in BMS-582664 VEGFR inhibitor MCF-7, MCF 7/adr, MX or topotecan in S1, S1 80 M1 cells, were each added, mitoxantrone and cisplatin in HEK293/pcDNA3.1, ABCG2 482 R5, Dai et al. Page 3 Cancer Res Author manuscript, increases available in PMC 2009 1 October. PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript NIH ABCG2 ABCG2 482 G2 and 482-T7 cells. The concentrations required to inhibit growth of 50% was calculated from the survival curves using the method of Bliss.
The Widerstandsf Conductivity was calculated by dividing the IC50 for MDR cells that calculates the sensitive parental cells. The degree of resolution and high MDR was calculated by dividing the IC50 for cells with the chemotherapeutic agent in the absence of lapatinib that have been obtained in the presence of lapatinib. Athymic mice Nacktm animals, 5 g of 6 weeks and weighing from 18 23 were used for the cell xenografts KBv200. All animals were sterilized food and water. MDR human carcinoma xenograft model of xenograft cells KBv200 prepared as described by Chen et al. Briefly, cells cultured in vitro KBv200 harvested and injected subcutaneously into mice in the shoulder Nacktm Implanted. When the tumors reached a mean diameter of 0.
5 cm, were Mice were randomized into 4 groups and treated with a NEN of the following Pl: A saline solution, paclitaxel 2, 3 lapatinib, lapatinib and paclitaxel fourth The K body weight Of the animal was measured every 3 days to adjust medication dose. The two perpendicular diameters recorded every 3 days, and tumor volume was calculated using the formula: The curve of tumor growth relative to the volume of tumor implantation. The Mice have been to Sthesiert and get tet When the mean tumor weight greater than 1 g in the control group. The tumor tissue was obtained from M Mice removed and weighed. The inhibition was calculated using the formula: the accumulation of doxorubicin and mitoxantrone intracellular accumulation of doxorubicin in MCF re ABCB1overexpessing 7/adr their parents and sensitive MCF-7 cells was analyzed by flow cytometry. Logarithmically growing cells were treated with 0.625, 1.25 or 2.5 M lapatinib at 37 for 3 h. Then, 10 M doxorubicin was added to the medium and incubation for a further 3 h