HIF－1α@Fe3O4纳米颗粒标记缺氧条件下胰腺癌PANC1细胞及其MRI表现  

HIF-1α@ Fe3O4 labeled pancreatic cancer PANCl cells under hypoxia and its MRI detection

作　　者：谢晓东[1] 王冬青[2] 张磊[1] 宋廉[2] 李丹 沈文荣[1] 

机构地区：[1]江苏省肿瘤医院影像科,南京210009 [2]江苏大学附属医院影像科

出　　处：《中华胰腺病杂志》2017年第5期326-329,共4页Chinese Journal of Pancreatology

基　　金：江苏省肿瘤医院院级课题（ZN201611）;江苏省社会发展项目（BE2015668）

摘　　要：目的探讨新型纳米颗粒缺氧诱导因子－1α（HIF－1α）螯合四氧化三铁探针（HIF－1α@Fe3O4）标记胰腺癌PANC1细胞的可行性及其3.0T MRI扫描时T2WI信号的改变。方法在缺氧条件下培养胰腺癌PANC1细胞，采用蛋白质印迹法检测细胞HIF－1α及干细胞标志蛋白CD133、Oct－4、Sox－2的表达。收集缺氧培养24 h的PANC1细胞，并分别与5、15、45 μg/ml的HIF－1α@Fe3O4纳米颗粒共孵育24 h，检测HIF－1α@Fe3O4标记的PANC1细胞数及细胞存活率，采用3.0T MRI扫描记录细胞在T2WI图上的信号强度。结果在缺氧培养下PANC1细胞HIF－1α表达量较常氧培养组明显增加，且随着缺氧时间的延长进一步增加（P值均〈0.05），干细胞标志蛋白CD133、Oct－4、Sox－2的表达与HIF－1α表达呈正相关。PANC1细胞与不同浓度HIF－1α@Fe3O4共孵育24 h后，胞质内蓝色铁染颗粒细胞呈浓度依赖性增多，45 μg/ml浓度时最高，达100%。常氧培养组、缺氧培养未标记组及缺氧培养45 μg/ml HIF－1α@Fe3O4标记组的细胞存活率分别为（87.0±2.1）%、（84.7±2.7）%、（85.0±3.8）%，差异无统计学意义（P〉0.05）。通过3.0T MRI扫描，未标记组及5、15、45 μg/ml HIF－1α@Fe3O4标记组细胞T2WI的信号强度比分别为1.017±0.046、0.793±0.041、0.447±0.032、0.240±0.031，各组间差异有统计学意义（F＝80.0，P〈0.05）。结论缺氧有利于胰腺癌PANC1细胞干细胞特性的维持。HIF－1α@Fe3O4探针可成功标记缺氧培养下高表达HIF－1α的PANC1细胞，且标记细胞在3.0T MRI扫描的T2WI图上的信号强度减弱。ObjectiveTo explore the feasibility of novel nano-particle HIF-1α@Fe3O4 labeled pancreatic cancer PANC1 cells as well as the changes of signal intensity in 3.0T MRI scan.MethodsPancreatic cancer PANC1 cells were cultured in hypoxia condition, and hypoxia-inducible-factor-1α（HIF-1α） and stem cell markers CD133, Oct-4, Sox-2 were detected by Western blot assay. Cells cultured under hypoxia for 24 h were collected and then co-incubated with 5, 15 and 45 μg/ml HIF-1α@Fe3O4 for 24 h. The number of HIF-1α@Fe3O4 labeled PANC1 cells and cell survival rate were detected, and the signal intensity of T2WI image for PANC1 cells was measured by a 3.0T MRI system.ResultsIn hypoxia condition, HIF-1α level was obviously increased compared with that of normoxic culture, which was further increased with the increase of hypoxia time（all P〈0.05） . Stem-cell markers CD133,Oct-4 and Sox-2 was positively correlated with HIF-1α level. Co-cultured with different concentrations of HIF-1α@Fe3O4 for 24 h, blue-stained iron particles in cytoplasm of PANC1 cells was dosage-dependently increased, and the peak was at the concentration of 45 μg/ml, which could reach 100%. The survival rate of the PANC1 cells cultured in normoxic condition, the unlabeled and labeled in hypoxic condition group were（87.0±2.1）%, （84.7±2.7）% and （85±3.8）%, respectively, and the difference was not statistically significant（P〉0.05）. In 3.0T MRI scan, T2WI signal intensity in unlabeled group and 5, 15 and 45 μg/ml labeled group was 1.017±0.046, 0.793±0.041, 0.447±0.032 and 0.240±0.031, and the difference was not statistically significant（F=80.0, P〉0.05）.ConclusionsHypoxia condition could promote and maintain the stemness in PANC1 cells. HIF-1α@Fe3O4 probe could successfully label HIF-1α highly expressed PANC1 cells during hypoxia condition, and a significant decrease in T2WI signal intensity can be detected by a 3.0T MRI system.

关 键 词：胰腺肿瘤 缺氧诱导因子-1Α 肿瘤干细胞 磁共振成像 

分 类 号：R735.9[医药卫生—肿瘤] R445.2[医药卫生—临床医学]