胃肠道间质瘤伊马替尼耐药细胞株的建立及细胞凋亡的实验研究
投稿时间:2016-07-26  修订日期:2016-11-05  点此下载全文
引用本文:顾彬彬,叶丽萍,方从诚,王俊,王超,吴坚芬.胃肠道间质瘤伊马替尼耐药细胞株的建立及细胞凋亡的实验研究[J].医学研究杂志,2017,46(6):117-120
DOI: 10.11969/j.issn.1673-548X.2017.06.030
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作者单位E-mail
顾彬彬 317000 台州医院消化内科  
叶丽萍 317000 台州医院消化内科 yelp@enzemed.com 
方从诚 317000 台州医院消化内科  
王俊 317000 台州医院消化内科  
王超 317000 台州医院消化内科  
吴坚芬 317000 台州医院消化内科  
基金项目:浙江省台州市科技计划项目(131ky13)
中文摘要:目的 体外构建胃肠道间质瘤耐药细胞株GIST-882-IM,鉴定其生物学特性。方法 采用首剂大剂量冲击和逐步增加剂量相结合的方法诱导胃肠道间质瘤细胞GIST-882对Imatinib耐药,建立耐药细胞株,CCK8法绘制细胞生长曲线,计算细胞的倍增时间及半抑制浓度IC50,流式细胞仪检测GIST-882及耐药株GIST-882-IM细胞凋亡及细胞周期。结果 耐药株GIST-882-IM的倍增时间为58.95±1.56h,高于细胞株GIST-882 49.49±0.69h,GIST-882-IM IC50=1.869±0.103μmol,GIST-882 IC50=0.054±0.001μmol,其耐药指数为34.62(P<0.01)。耐药细胞株GIST-882-IM相对于GIST-882,凋亡受到抑制,24、48、72h其细胞凋亡率分别为1.97%±0.38%、2.87%±0.38%、4.40%±0.36%,GIST-882凋亡率分别为6.07%±0.96%、11.17%±0.64%、16.97%±0.12%。细胞周期分析,耐药细胞株的G0/G1期增加,S期、G2/M期减少。结论 成功构建GIST882 Imatinib耐药细胞株,耐药细胞与非耐药细胞的生物学特性不同。这为下一步研究伊马替尼耐药机制和筛选伊马替尼耐药的替代靶向药物奠定基础。
中文关键词:耐药细胞株  细胞生长曲线  耐药指数  细胞凋亡
 
Establishment and Apotosis of an Imatinib Resistant Cell Line of Gastrointestinal Stromal Tumor
Abstract:Objective To establish the imatinib-resistant gastrointestinal stromal cell line (GIST-882-IM) and explore its biological characteritics. Methods Imtinib-resistant cell line(GIST-882-IM) was established by the high concentration Imatinib. The cell growth curve,doubling time and IC50 was calculated by CCK-8 assay. The apoptosis and the cell cycle of GIST-882、GIST-882-IM was tested by flow cytometry. Results The doubling time of GIST-882-IM and GIST-882 was 58.95±1.56h and 49.49±0.69h respectively (P<0.01).IC50 for GIST-882-IM and GIST-882 was IC50=1.869±0.103μmol and 0.054±0.001μmol respectively (P<0.01).The resistance of GIST-882-IM to Imatinib was 34.62 fold of GIST-882. Compared with GIST-882,apotosis of the GIST-882-IM was inhibited. The apotosis rate of GIST-882-IM in 24,48,72h was 1.97%±0.38%,2.87%±0.38%,4.40%±0.36% while the GIST-882 was 6.07%±0.96%、11.17%±0.64%、16.97%±0.12% respectively (P<0.01).The cell numbers of G0/G1 phase increased while decreased in S and G2/M phase in GIST-882-IM cell. Conclusion The Imatinib-resistant gastrointestinal stromal cell line GIST-882-IM was successful established. The GIST-882 and GIST-882-IM cells were different in biological characteristics.The GIST-882-IMcell will be uesd to sutdy the mechanism of Imatinib resistance in GIST and search another targed drug to eliminate the drug resistance.
keywords:Drug-resistant cell line  Cell growth curve  Resistance  Apotosis
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