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- W3043809546 abstract "1493 Irinotecan (CPT-11), a camptothecin analog, is converted to the active topoisomerase I inhibitor, SN-38, by carboxylesterase. CPT-11 is widely used for the treatment of advanced colorectal carcinoma. CPT-11 refractory colorectal cancer patients have shown benefit by treatment with anti-EGFR mAb, cetuximab, when given alone or in combination with CPT-11. In order to study current and novel treatment options for CPT-11 refractory colorectal cancer, two CPT-11 resistant colorectal cancer cell sublines were isolated from the parental cell line, SW-948. Two methods were used to develop sublines resistant to CPT-11. First, SW-948 cells were initially exposed to a high dose (2-fold higher than the IC50 dose) of CPT-11 to obtain the resistant cell line, SW948CPTH. Second, SW-948 cells were initially exposed to a low dose (4-fold lower than the IC50 dose) of CPT-11 to obtain the resistant cell line, SW948CPTL. Both methods used a stepwise increase and continuous exposure to CPT-11 on the cells. The cells were monitored for drug resistance by clonogenic assay. Upon attaining an increased resistance of ≥10-fold in the IC50 dose of CPT-11compared to the parental cell line, the drug was removed and the cells were monitored for retention of CPT-11 resistance for >100 days. Cross-resistance testing to other topoisomerase I inhibitors showed increased resistance to CPT (SW948CPTH) and SN-38 (SW948CPTH and SW948CPTL) but not lapachone (SW948CPTH). Cross-resistance studies to other chemotherapeutic agents showed increased resistance to cisplatin and etoposide by SW948CPTL but not 5-FU by either subline. Microarray analyses showed the differential expression of 464 (p≤0.01) or 1488 (p≤0.05) genes when comparing the SW948CPTH to SW-948 cells and 27 (p≤0.01) or 333 (p≤0.05) genes when comparing the SW948CPTL to SW-948 cells. Ninety-eight (p≤0.05) genes were identified that were differentially expressed in both SW948CPTH and SW948CPTL cells compared to the parental cell line. SW948CPTL cells were found to overexpress the p-glycoprotein (p-gp) gene and protein by microarray and FACS analyses, respectively. Verapamil was shown to increase the intracellular concentration of CPT-11 in SW948CPTL cells. SW948CPTH cells were found to overexpress the single-strand break repair gene, XRCC1, and protein by microarray analysis and immunoblot, respectively. Immunoblot analysis indicated the EGFR expression was similar in the parental cell line and each subline, however erbB-2 gene and protein expression was increased in SW948CPTH cells. Exposure of the parental cell line and each subline to cetuximab was shown to reduce the expression of XRCC1 protein in SW948CPTH and SW948CPTL cells. The mechanism of CPT-11 drug resistance in SW948CPTL cells involves p-gp whereas the mechanism of CPT-11 drug resistance in SW948CPTH cells may involve one or more genes, one of which may be the XRCC1 gene." @default.
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- W3043809546 date "2005-05-01" @default.
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- W3043809546 title "Development and analyses of CPT-11 resistant sublines of human colorectal adenocarcinoma cell line, SW-948" @default.
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