ZHANG Ping,ZHOU Zhi-guo,GAO Xian-shu,et al.Isolation and characterization of radioresistant human esophageal cancer cells and the differential gene expression by cDNA microarray analysis[J].Chinese Journal of Radiological Medicine and Protection,2006,26(6):566-570 |
Isolation and characterization of radioresistant human esophageal cancer cells and the differential gene expression by cDNA microarray analysis |
Received:December 22, 2005 |
DOI: |
KeyWords:Esophageal cancer cell lines Radioresistance Cell cycle cDNA microarray analysis Flow cytometry |
FundProject:国家自然科学基金资助项目(30240057);河北省自然科学基金资助项目(303508) |
Author Name | Affiliation | E-mail | ZHANG Ping | Department of Radiation Oncology, Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011, China | xsgao777@hotmail.com | ZHOU Zhi-guo | Department of Radiation Oncology, Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011, China | | GAO Xian-shu | 北京大学第一医院放疗科 | | 卢付河 | Department of Radiation Oncology, Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011, China | | 乔学英 | Department of Radiation Oncology, Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011, China | | 宋永辉 | Department of Radiation Oncology, Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011, China | |
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Abstract:: |
Objective To observe the differential gene expression in esophageal cancer cell with different radiosensitivity by cDNA microarray analysis. Methods A radioresistant cell line, TE13R120, was isolated and characterized in vitro from a human esophageal cancer cell line TE13 by repeated 60Co gamma-ray irradiation, 120 Gy in total. Morphological changes from TE13 to TE13R120 were observed by phase-contrast microscopy. The relative radiosensitivities of tumor cells were assessed by standard colony formation assays. The time-course of cell cycle distribution after irradiation was investigated by flow cytometry (FCM). The differential gene expression was identified using cDNA microarray technique. Results The population doubling time of TE13 and TE13R120 was 33.94 h and 39.93 h, respectively. For TE13R120 cells, D0 of radiation sensitivity was 2.85 Gy, while D0 was 1.63 Gy for TE13 cells. The surviving fractions at 2 Gy (SF2) were 0.64 for TE13R120 cells and 0.55 for TE13 cells. Cell cycle distributions were different for TE13 cells and TE13R120 cells 12-48 h after irradiation. A marked accumulation of G1 phase was observed after 4 Gy irradiation for TE13 cells, and reached a peak at 12 h after irradiation. However, no apparent change of cell cycle distribution for TE13R120 cells was observed after irradiation and only a slight accumulation of cells in G2/M phase was observed during the period from 12 h to 24 h after irradiation. By the use of cDNA microarray analysis of differential gene expression between TE13R120 cell line and its parental TE13 cell line, 96 upregulated genes in TE13R120 cells and 80 down-regulated genes were found. Conclusions A radioresistant variant cell line, denoted by TE13R120, from the TE13 cell line was successfully isolated by repeated gamma-radiation exposures. Significant changes in gene expressions were observed in this variant cell line. A marked arrest of cells in the G1 phase was observed in TE13 cells after 4 Gy irradiation with, but no apparent change of cell cycle distribution for TE13R120 cells. |
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