高低转移潜能肝癌在骨髓间充质干细胞干预前后的Gd-RGD成像  

Hepatocellular carcinoma with high and low metastatic potential before and after bone marrow mesenchymal stem cell transplantation: a MRI imaging study using Gd-RGD probe

作　　者：李天然[1] 黄晓斌 杨志杰 卢光明 李延军 禹名卉 Li Tian-ran;Huang Xiao-bin;Yang Zhi-jie;Lu Guang-ming;Li Yan-jun;Yu Ming-hui(Department of Radiology,No.1 Affiliated Hospital of Chinese PLA General Hospital,Beijing 100048,China;Department of Radiology,Chinese PLA 95th Hospital,Putian 351100,Fujian Province,China;Department of Radiology,Chinese PLA Nanjing General Hospital,Nanjing 210000,Jiangsu Province,China)

机构地区：[1]解放军总医院第一附属医院放射科,北京市100048 [2]解放军第九五医院放射科,福建省莆田市351100 [3]解放军南京总医院医学影像科,江苏省南京市210000

出　　处：《中国组织工程研究》2018年第21期3341-3348,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81271607);国家博士后基金(2015M572810)~~

摘　　要：背景:随着现代分子影像学的进步,使骨髓间充质干细胞干预肿瘤效能的实时监测成为可能。目的:构建新型转移相关MRI分子影像探针Gd-RGD,通过MRI成像在体观察骨髓间充质干细胞干预前后肝癌转移和增殖行为的变化。方法:构建MR转移相关因子分子影像探针整合素α_vβ_3配体cRGD-PEG-DGL-DTPA-Gd(Gd-RGD),并进行~1H-MRS及ICP-AES分析鉴定。制作高转移潜能MHCC97-H和低转移潜能MHCC97-L肝癌动物模型,尾静脉注射骨髓间充质干细胞悬液干预6周,计算肿瘤质量抑制率。骨髓间充质干细胞干预后分别用Gd-DTPA和Gd-RGD两种示踪剂进行MRI成像实验,以SNR和CNR为半定量指标。将MHCC97-H和MHCC97-L与骨髓间充质干细胞进行Transwell共培养,置于37℃,体积分数为5%CO_2培养箱内培养48 h后采用qPCR法检测转化生长因子β1、骨桥蛋白、整合素亚单位α_v和β_3的表达。结果与结论:(1)经骨髓间充质干细胞干预后,肝癌肿瘤组织质量明显降低。在第3周时,肿瘤质量抑制率最高;(2)对于高转移潜能肝癌(MHCC97-H),骨髓间充质干细胞干预后Gd-RGD成像SNR和CNR低于骨髓间充质干细胞干预前(P<0.05)。对于低转移潜能肝癌(MHCC97-L),骨髓间充质干细胞干预前后Gd-RGD成像SNR比较差异无显著性意义(P>0.05),骨髓间充质干细胞干预后CNR低于骨髓间充质干细胞干预前(P<0.05);(3)骨髓间充质干细胞干预后Gd-RGD成像SNR在高、低转移潜能肝癌之间差异有显著性意义(P<0.05);Gd-DTPA成像SNR和CNR差异不如Gd-RGD成像差异显著;(4)高转移潜能细胞(MHCC97-H)与骨髓间充质干细胞共培养后,骨桥蛋白、整合素亚单位β_3、转化生长因子β1表达有明显的下降(P<0.05),而α_v表达无明显变化(P>0.05)。低转移潜能细胞(MHCC97-L)与骨髓间充质干细胞共培养后,骨桥蛋白、β_3、转化生长因子β1、α_v表达均下降(P<0.05);(5)结果表明,MRI成像中Gd-DTPA和Gd-RGD示踪剂CNR指标可以很好地区BACKGROUND： With the advancement of modern molecular imaging, possibilities to monitor the efficacy of bone marrow mesenchymal stem cells （BMSCs） against tumors have been achieved.OBJECTIVE： To construct a novel metastasis-associated MRI molecular imaging probe, Gd-RGD, and to observe metastasis and proliferation of hepatocellular carcinoma （HCC） before and after intervention with BMSCs using MRI imaging.METHODS： The metastasis-associated MR molecular imaging probe, integrin αvβ3 ligand cRGD-PEG-DGL-DTPA-Gd （Gd-RGD）, was constructed, and the ingredient analysis and identification were implemented through 1H-magnetic resonance spectroscopy （1H-MRS） and inductively coupled plasma-atomic emission spectrometry （ICP-AES） methods. MHCC97-H and MHCC97-L animal models were established. After BMSCs intervention was administrated via the tail vein for 6 weeks, tumor weight inhibition rate was calculated. MRI imaging with a metastasis-associated integrin ligand RGD molecular probe was performed. Molecular imaging probe Gd-DTPA was taken as control. Signal noise ratio （SNR） and contrast to noise ratio （CNR） of MRI were taken as semi-quantitative indicators. MHCC97-H and MHCC97-L animal models before BMSCs intervention were co-cultured with BMSCs using Transwell method. Quantitative PCR method was used to detect proliferation and metastasis-associated indicators, tumor growth factor β1 （TGF-β1）, osteopontin, integrin subunits αv and β3.RESULTS AND CONCLUSION： （1） After BMSCs transplantation, the tumor tissue weight of HCC decreased obviously, and the tumor weight inhibition rate was peaked in the 3rd week. （2） For high metastatic potential （MHCC97-H）, both SNRs and CNRs of Gd-RGD MR imaging before and after BMSCs intervention were statistically different （P 〈 0.05）. For low metastatic potential （MHCC97-L）, SNRs of Gd-RGD MR imaging before and after BMSCs intervention were not statistically different （P 〉 0.05）, while CNRs were statistically different �

关 键 词：骨髓间充质干细胞 高转移潜能肝癌细胞 低转移潜能肝癌细胞 整合素 Gd-RGD GD-DTPA 干细胞 国家自然科学基金 骨髓 间质干细胞移植 肝肿瘤 整合素αVβ3 磁共振成像 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]