氯化钴对人胰腺癌耐药细胞株Patu8988/5-FU的线粒体损伤及其耐药性的影响  

Effects of cobalt chloride on mitochondrial damage and drug resistance of 5-FU-resistant human pancreatic cancer Patu8988/5-FU cell line

作　　者：宋毕清[1] 时开网[1] 杨士勇[1] 

机构地区：[1]南京医科大学附属南京第一医院普通外科,江苏南京210006

出　　处：《南京医科大学学报（自然科学版）》2012年第6期811-815,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省自然科学基金(H200945)

摘　　要：目的:氯化钴(CoCl2)作用于人胰腺癌耐药细胞株Patu8988/5-FU致其化学性缺氧,观察其线粒体损伤及耐药性的影响。方法:采用梯度浓度CoCl2(0、6.25、12.50、25.00、50.00、100.00、200.00、400.00μmol/L)培养人胰腺癌耐药细胞株Patu8988/5-FU,MTT法测定Patu8988/5-FU细胞增殖活性和耐药性;JC-1荧光染色流式细胞仪分析线粒体膜电位变化;RT-PCR检测HIF-1α及多药耐药基因MDR1的表达情况;罗丹明外排实验检测Patu8988/5-FU细胞膜上的P-gp泵功能。结果:随着CoCl2浓度升高或以100μmol/L作用时间延长,Patu8988/5-FU增殖活性下降,线粒体膜电位去极化增加。常氧和缺氧时其对5-氟脲嘧啶(5-Fu)的耐药指数分别达到28.11±3.19、52.08±3.53(P<0.01);HIF-1α和MDR1 mRNA的表达亦明显高于常氧对照组(P<0.05)。罗丹明在缺氧细胞株内的积聚量低于常氧对照组(P<0.01)。结论:利用CoCl2模拟Patu8988/5-FU化学性缺氧存在浓度和时间依赖性;缺氧使Patu8988/5-FU获得性耐药性增加可能是通过引起HIF-1α表达增加和上调多药耐药基因MDR1的表达引起的。Objective：To investigate the effects of mitochondrial damage and drug resistance of 5-FU-resistant human pancreatic cancer cell line Patu8988/5-FU cultured by cobalt chloride （COC12）. nethods：Patu8988/5-FU were cultured in medium containing different concentrations of COC12 （0,6.25μmol/L, 12.50 μmol/L, 25.00 μmol/L, 50.00 μmol/L, 100.00 μmol/L, 200.00μmol/L, 400.00 μmol/L）. MTT assay was used to detect the cell proliferation activity and drug resistance of Patu8988/5-FU. JC-1 fluorescence staining with flow cytometry was used to analyze the mitochondria membrane potential. The mRNA expressions of HIF-1α and MDR1 were detected by RT-PCR technique. P-gp pump function of Patu8988/5-FU cells＇ membrane was tested by Rh123. Results： With COC12 concentration rising or 100μmol/L COC12 treatment time extension,the proliferation activity of Patu8988/5-FU cells was degressive, the depolarization of mitochondrial membrane potential was increased. Drug resistance index of Patu8988/5-FU to 5-FU in normoxia and hypoxia reaches to 28.11±3.19 and 52.08 ± 3.53,respectively （P 〈 0.01）. The mRNA expressions of HIF-1α and MDR1 in Patu8988/5-FU cells is higher in hypoxia than in normoxia （P 〈 0.01 ）. The accumulation amount of Rh123 in cytoplasm of Patu8988/5-FU cells was significantly lower in hypoxia than in normoxia（P 〈 0.01）. Conclusion： COC12 induced chemistry hypoxia of Patu8988/5-FU cells in a time-and-dose dependent manner. CoCI： mimicing hypoxia dramatically increased resistance of Patu8988/5- FU cells to 5-FU, and maybe through upregulating the expression of HIF-1α and MDR1.

关 键 词：缺氧 胰腺癌 多药耐药 

分 类 号：R735.9[医药卫生—肿瘤]