重庆医科大学附属第二医院妇产科
Published:2014,
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Yin Mingyue, Sun Jiangchuan, Chang Shufang, et al. Comparison on hypoxia induced drug resistance model for two kinds of human ovarian cancer cell lines. [J]. Journal of Chongqing Medical University 38(11):1533-1536(2014)
目的:采用二氯化钴(cobaltous chloride,CoCl2)化学诱导法构建人卵巢癌细胞株A2780和SKOV3的乏氧耐药模型,并比较二者耐药特性。方法:(1)体外培养的A2780和SKOV3细胞分别加入不同浓度CoCl2培养液作用12、24、48、72 h,采用MTT法检测其增殖活性及对紫杉醇的耐药倍数;(2)采用150 μmol/L CoCl2培养液作用24 h构建乏氧环境,RT-PCR检测常氧和乏氧条件下乏氧诱导因子(hypoxia inducible factor,HIF)1α mRNA的表达情况。结果:2种细胞的增殖抑制率与CoCl2呈剂量-时间依赖关系(A2780组:F=1 165.416,P=0.000;SKOV3组:F=2 802.394,P=0.000)。随着CoCl2浓度的增加,A2780和SKOV3细胞对紫杉醇的耐药倍数增加(F=7 842.711,P=0.000);在相同CoCl2浓度作用下,SKOV3细胞对紫杉醇的耐药倍数明显高于A2780(50 μmol/L组:t=-48.287,P=0.000;100 μmol/L组:t=-263.205,P=0.000;150 μmol/L组:t=-143.305,P=0.000)。150 μmol/L CoCl2作用24 h,2种细胞耐药倍数分别为(9.84±0.11)和(21.08±0.08),并出现稳定的HIF-1α mRNA表达,其中SKOV3细胞HIF-1α mRNA的表达增加量明显高于A2780细胞(t=-5.573,P=0.000)。结论:CoCl2化学诱导法可成功建立卵巢癌细胞乏氧耐药模型,其耐药倍数与细胞类型有关。在相同条件下,SKOV3细胞株乏氧耐药模型较A2780更易建立,是乏氧耐药逆转研究较为理想的细胞株。
Objective:To establish hypoxia drug resistance model(HDRM) of human ovarian cancer cell lines A2780 and SKOV3 by chemical induction method of Cobalt Chloride(CoCl2) and to compare the drug resistant characteristics of those two cell lines. Methods:CoCl2 with different concentrations(0,50,100,150,200,300 μmol/L) was added to A2780 and SKOV3 cells for 12,24,48,72 h. Cell proliferation activity(CPA) was detected by methyl thiazolyl tetrazolium(MTT). MTT was used to detect the drug resistance multiple(DRM) of paclitaxel under the different concentrations of CoCl2. The same cell lines without CoCl2 were taken as normoxic control. CoCl2 with concentration of 150 μmol/L was added to A2780 and SKOV3 cells medium for 24 h respectively and the expression of HIF-1α mRNA was detected by the real time polymerase chain reaction. The same cell lines without CoCl2 were taken as normoxic control. Results:The proliferation activity of A2780 and SKOV3 cells was correlated with CoCl2 in a dose and time dependent manner(A2780 group:F=1 165.416,P=0.000;SKOV3 group:F=2 802.394,P=0.000). Paclitaxel resistance fold of those two kinds of cells was increased with the increase of CoCl2 concentration(F=7 842.711,P=0.000). With the same concentration of CoCl2,DRM of SKOV3 cells was significantly higher than that of A2780 cells(50 μmol/L group:t=-48.287,P=0.000;100 μmol/L group:t=-263.205,P=0.000;150 μmol/L group:t=-143.305,P=0.000). With 150 μmol/L CoCl2 culturing for 24 h,DRM of A2780 and SKOV3 cells reached to 9.84±0.11 and 21.08±0.08,respectively. The increase of HIF-1α mRNA expression in SKOV3 cells was significantly higher than that of A2780 cells(t=-5.573,P=0.000). Conclusion:Ovarian cancer HDRM can be successfully established by CoCl2 induction method and DRM is related with cell characteristics. Under the same condition,SKOV3 cell line is easier to establish HDRM than A2780 cell line,and SKOV3 is an ideal cell line for hypoxia resistance reversal study.
乏氧耐药二氯化钴人卵巢癌细胞乏氧因子
hypoxia drug resistance modelcobalt chloridehuman ovarian cancer cellhypoxia inducible factor
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