E ABCC1 overcome ABCB1 and ABCG2 in vitro resistance to mediation relatively low concentrations. Materials and Methods Plant material and isolation of the compound. The aerial parts of Hyptis verticillata Jacq were collected in the summer of 2007 in Kingston, Jama and voucher specimens were deposited in the herbarium of the University Cyclooxygenas of t the West Indies, Mona, Jama get off know that methanolic extracts from aerial parts of the plant was distributed between water and diethyl ether. The ether extract was further between methanol / water and n-hexane, methanol w Ssrige layer subjected to silica gel flash chromatography and 4 methoxy 9 8.9 6 dihydrofuronaphthodioxol that eluted from the fractions obtained partitioned with hexane / ethyl acetate and purification by HPLC high octadecyl silane.
For comparison, the standard chemotherapy drugs etoposide and mitoxantrone from Sigma Aldrich. Cells. S1T, K3T and KaT01F ATL cell lines in our laboratory were maintained in RPMI 1640 erg complements With 10% Fetal K F CX-4945 Protein kinase PKC inhibitor calf serum, 100 U / ml penicillin G, 100 g / ml streptomycin and 2 mM L glutamate and divide every 2 to 3 days, corresponding to the logarithmic growth phase, Jurkat, lymphoblasto human T-cells Cell line HL60 and human Promyelozytenleuk Mie cell line were kept the same. The line of c Lon human cell adenocarcinoma, lung non-small cell lung cancer cell line is, cancer cells Epidemo Of resistant or transfected fa Stable ABCC1 cDNA Leuk Chemistry cells or 2 KBG express Pgp and erythromyeloblastoid or resistant transfected fa is connected to the ABCG2 cDNA were kept in the same medium, and sp ter washed again culture flasks with trypsin and before use in stable assays.
The first KB M Rz cell line was kindly provided by Professor Shin ichi Akiyama of the University JNJ-26481585 of t Tokushima, 2 and KBG KBABCC1 Professor Ueda of Kyoto University Kazumitsu, and K562 and Yoshikazu Sugimoto K562/ABCG2 Professor of Keio University provided t. Cytotoxicity Tstest. To the cytotoxic effects of MTDND 4, 1104 cells / well in at least three times the assessment were in a 96-well flat-bottom-incubated, without or in the presence of serial dilutions of the compound in a humidified incubator at 37 C. The cells were harvested after 72 h and 2.
5 reactive diphenyl tetrazolium bromide was added to each well and incubated for 4 hours followed by resolution and high formazan crystals dried with sodium sulfate and 20% after dodecyl What the plates were left alone in the dark at least 2 hours before reading the absorbance at 540/630 nm on a microplate reader. Untreated cells were assigned a value of 100% Lebensf Ability, and the Lebensf Ability of the treated cells were expressed relative to untreated controls. 8 The WST-test was used to MDR cells to compare their contr Them. Ver apoptotic changes In cell morphology. Leuk Mie cells were incubated S1T absence or presence of 4 MTDND at various concentrations in a 24-well microtiter plate in a humidified incubator at 37 C for 48 h. The cells were then recovered and followed in glass slides hunter with a Cytospin device, from May Grunwald Giemsa. Cell cycle analysis. The cells were incubated S1T absence or presence of 4 MTDND at various concentrations in triplicate in at least a 24