A Direct and Sensitive Method for Determination of 5-Fluorouracil in Colorectal Cancer Cells: Evaluating the Effect of Stromal Cell on Drug Resistance of Cancer Cells
Fibroblasts in the stroma play a critical role in tumor evolution. In this study, we assessed the influence of colonic fibroblasts on colon cancer cells treated with 5-fluorouracil (5-FU), and mouse colon cancer cell lines MC38 and colonic fibroblasts NIH3T3 were used in this study. A sensitive and...
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2021-01-01
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| Series: | Journal of Analytical Methods in Chemistry |
| Online Access: | http://dx.doi.org/10.1155/2021/6689488 |
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| author | Fengjing Xu Zhipeng Wang Xinhua Song Mengwei Zhang Lili Cui Yanping Liu Hongxia Yan Shouhong Gao Yan Liu Wansheng Chen |
| author_facet | Fengjing Xu Zhipeng Wang Xinhua Song Mengwei Zhang Lili Cui Yanping Liu Hongxia Yan Shouhong Gao Yan Liu Wansheng Chen |
| author_sort | Fengjing Xu |
| collection | DOAJ |
| description | Fibroblasts in the stroma play a critical role in tumor evolution. In this study, we assessed the influence of colonic fibroblasts on colon cancer cells treated with 5-fluorouracil (5-FU), and mouse colon cancer cell lines MC38 and colonic fibroblasts NIH3T3 were used in this study. A sensitive and rapid UHPLC-MS/MS method for the quantitation of 5-FU from the cell and their medium has been successfully developed and validated. The cells were lysed with methanol, and the mixture was evaporated and then redissolved to extract intracellular 5-FU. The analysis was performed on UHPLC-MS/MS using an Atlantis T3-C18 column (3 μm, 2. 1 ∗ 100 mm) and gradient elution with acetonitrile and 0.1% formic acid in water. Method validation included the following parameters: the matrix effect range 88.82%–93.64% and the recovery range 93.52%–94.56%. The intraday and interday precision and accuracy were <11% and within ±6%, and the stability, specificity, carry-over, dilution effect, and linearity all conformed to the criteria. The method was applied to detect the concentration of 5-FU inside cells and cell culture medium. The preliminary results present that NIH3T3 could enhance the drug resistance of MC38 to 5-FU with a decreased intracellular concentration of 5-FU in MC38, which showed a positive relationship with NIH3T3 number. |
| format | Article |
| id | doaj-art-358d13d343224ae78e540ef713f15dbf |
| institution | Kabale University |
| issn | 2090-8865 2090-8873 |
| language | English |
| publishDate | 2021-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Analytical Methods in Chemistry |
| spelling | doaj-art-358d13d343224ae78e540ef713f15dbf2025-02-03T01:05:28ZengWileyJournal of Analytical Methods in Chemistry2090-88652090-88732021-01-01202110.1155/2021/66894886689488A Direct and Sensitive Method for Determination of 5-Fluorouracil in Colorectal Cancer Cells: Evaluating the Effect of Stromal Cell on Drug Resistance of Cancer CellsFengjing Xu0Zhipeng Wang1Xinhua Song2Mengwei Zhang3Lili Cui4Yanping Liu5Hongxia Yan6Shouhong Gao7Yan Liu8Wansheng Chen9College of Traditional Chinese Medicine, Yunnan University of Chinese Medicine, Kunming, Yunnan 650500, ChinaDepartment of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaDepartment of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaDepartment of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaDepartment of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaDepartment of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaCollege of Traditional Chinese Medicine, Yunnan University of Chinese Medicine, Kunming, Yunnan 650500, ChinaCollege of Traditional Chinese Medicine, Yunnan University of Chinese Medicine, Kunming, Yunnan 650500, ChinaDepartment of Pharmacy, Xinhua Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, ChinaDepartment of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, ChinaFibroblasts in the stroma play a critical role in tumor evolution. In this study, we assessed the influence of colonic fibroblasts on colon cancer cells treated with 5-fluorouracil (5-FU), and mouse colon cancer cell lines MC38 and colonic fibroblasts NIH3T3 were used in this study. A sensitive and rapid UHPLC-MS/MS method for the quantitation of 5-FU from the cell and their medium has been successfully developed and validated. The cells were lysed with methanol, and the mixture was evaporated and then redissolved to extract intracellular 5-FU. The analysis was performed on UHPLC-MS/MS using an Atlantis T3-C18 column (3 μm, 2. 1 ∗ 100 mm) and gradient elution with acetonitrile and 0.1% formic acid in water. Method validation included the following parameters: the matrix effect range 88.82%–93.64% and the recovery range 93.52%–94.56%. The intraday and interday precision and accuracy were <11% and within ±6%, and the stability, specificity, carry-over, dilution effect, and linearity all conformed to the criteria. The method was applied to detect the concentration of 5-FU inside cells and cell culture medium. The preliminary results present that NIH3T3 could enhance the drug resistance of MC38 to 5-FU with a decreased intracellular concentration of 5-FU in MC38, which showed a positive relationship with NIH3T3 number.http://dx.doi.org/10.1155/2021/6689488 |
| spellingShingle | Fengjing Xu Zhipeng Wang Xinhua Song Mengwei Zhang Lili Cui Yanping Liu Hongxia Yan Shouhong Gao Yan Liu Wansheng Chen A Direct and Sensitive Method for Determination of 5-Fluorouracil in Colorectal Cancer Cells: Evaluating the Effect of Stromal Cell on Drug Resistance of Cancer Cells Journal of Analytical Methods in Chemistry |
| title | A Direct and Sensitive Method for Determination of 5-Fluorouracil in Colorectal Cancer Cells: Evaluating the Effect of Stromal Cell on Drug Resistance of Cancer Cells |
| title_full | A Direct and Sensitive Method for Determination of 5-Fluorouracil in Colorectal Cancer Cells: Evaluating the Effect of Stromal Cell on Drug Resistance of Cancer Cells |
| title_fullStr | A Direct and Sensitive Method for Determination of 5-Fluorouracil in Colorectal Cancer Cells: Evaluating the Effect of Stromal Cell on Drug Resistance of Cancer Cells |
| title_full_unstemmed | A Direct and Sensitive Method for Determination of 5-Fluorouracil in Colorectal Cancer Cells: Evaluating the Effect of Stromal Cell on Drug Resistance of Cancer Cells |
| title_short | A Direct and Sensitive Method for Determination of 5-Fluorouracil in Colorectal Cancer Cells: Evaluating the Effect of Stromal Cell on Drug Resistance of Cancer Cells |
| title_sort | direct and sensitive method for determination of 5 fluorouracil in colorectal cancer cells evaluating the effect of stromal cell on drug resistance of cancer cells |
| url | http://dx.doi.org/10.1155/2021/6689488 |
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