基于临床手术标本的胰腺癌原位移植模型建立及评价
|
摘要
目的建立基于临床肿瘤标本的胰腺癌原位移植模型,开展模型的评价研究。方法将临床新鲜的胰腺癌手术标本移植到裸鼠胰腺部位,建立原位异种移植模型(PDOX),通过活体成像技术和PET/CT对肿瘤生长状况进行评价;采用STR技术分析移植瘤的来源,通过组织形态观察和免疫组化判定移植瘤的病理学特征,同时检测PDOX模型外周血中CA19-9的表达水平。结果模型建立6周后,通过活体成像观察到近红外荧光信号明显富集在肿瘤部位,通过荧光强度可以初步判断肿瘤位置和大小;使用小动物PET/CT可清晰观察到腹部18F-FDG分子探针富集区域,与预期的肿瘤位置和大小一致;安死术后,解剖分离的各个脏器组织及肿瘤组织,通过近红外荧光成像进一步确认肿瘤生长状况;STR检测证实移植瘤人源性比率为99.99%;组织形态学和免疫组化分析表明移植肿瘤生长于裸鼠胰腺;血清中CA19-9检测进一步证实了胰腺癌的发生。结论成功建立了人胰腺癌原位异种移植模型,通过小动物活体成像、PET/CT等技术对模型进行了全面评估,为胰腺癌的发病机制和治疗研究提供了良好的动物模型。
Objective To establish and evaluate a patient-derived orthotopic xenograft( PDOX) model of pancreatic cancer. Methods Tissues of patient-derived pancreatic tumor were transplanted into nude mouse pancreas by surgery. The PDOX models were evaluated by the small animal near infrared fluorescence( NIRF) optical imaging and PET/CT. The traceability of PDOX models was detected by STR technology,and the pathological changes were observed by H&E staining,immunohistochemistry,and serum level of CA19-9 was detected by ELISA. Results Apparent NIRF were observed to be accumulated in pancreatic site by optical imaging system. The location and size of the xenografts tumor were revealed by fluorescence intensity. The PET/CT images with18 F-FDG molecular probe confirmed the tumor's location and size. Ex vivo NIRF imaging of isolated organ further showed the tumor formation. The traceability of PDOX models was99. 99% with human origin. H&E staining pathology and immunohistochemistry indicated the pancreatic cancer characteristics. The high serum level of ca19-9 confirmed the mice bearing tumor. Conclusions Pancreatic PDOX models are successfully established in this study,and it can be evaluated comprehensively by NIRF optical imaging and PET/CT,providing an appropriate platform for further research of pancreatic cancer.
Objective To establish and evaluate a patient-derived orthotopic xenograft( PDOX) model of pancreatic cancer. Methods Tissues of patient-derived pancreatic tumor were transplanted into nude mouse pancreas by surgery. The PDOX models were evaluated by the small animal near infrared fluorescence( NIRF) optical imaging and PET/CT. The traceability of PDOX models was detected by STR technology,and the pathological changes were observed by H&E staining,immunohistochemistry,and serum level of CA19-9 was detected by ELISA. Results Apparent NIRF were observed to be accumulated in pancreatic site by optical imaging system. The location and size of the xenografts tumor were revealed by fluorescence intensity. The PET/CT images with18 F-FDG molecular probe confirmed the tumor's location and size. Ex vivo NIRF imaging of isolated organ further showed the tumor formation. The traceability of PDOX models was99. 99% with human origin. H&E staining pathology and immunohistochemistry indicated the pancreatic cancer characteristics. The high serum level of ca19-9 confirmed the mice bearing tumor. Conclusions Pancreatic PDOX models are successfully established in this study,and it can be evaluated comprehensively by NIRF optical imaging and PET/CT,providing an appropriate platform for further research of pancreatic cancer.
引文
[1]Kamisawa T,Wood LD,Itoi T,et al.Pancreatic cancer[J].Lancet,2016,388(10039):73-85.
[2]Smith RA,Andrews KS,Brooks D,et al.Cancer screening in the United States,2017:a review of current American Cancer society guidelines and current issues in cancer screening[J].CA Cancer J Clin,2017,67(2):100-121.
[3]Siegel RL,Miller KD,Jemal A.Cancer statistics,2017[J].CA Cancer J Clin,2017,67(1):7-30.
[4]Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.
[5]Rosfjord E,Lucas J,Li G,et al.Advances in patient-derived tumor xenografts:from target identification to predicting clinical response rates in oncology[J].Biochem Pharmacol,2014,91(2):135-43.
[6]Murakami T,Murata T,Kawaguchi K,et al.Cervical cancer patient-derived orthotopic xenograft(PDOX)is sensitive to cisplatinum and resistant to nab-paclitaxel[J].Anticancer Res,2017,37(1):61-65.
[7]Kawaguchi K,Igarashi K,Murakami T,et al.Tumor-targeting Salmonella typhimurium A1-R sensitizes melanoma with a BRAFV600E mutation to vemurafenib in a patient-derived orthotopic xenograft(PDOX)nude mouse model[J].J Cell Biochem,2017,118(8):2314-2319.
[8]Rapic S,Vangestel C,Verhaeghe J,et al.Characterization of an orthotopic colorectal cancer mouse model and its feasibility for accurate quantification in positron emission tomography[J].Mol Imaging Biol,2017,20:1536-1632.
[9]Wang Z,Kang F,Gao Y,et al.Metformin promotes 2-deoxy-2-[18F]fluoro-D-glucose uptake in hepatocellular carcinoma cells through Fox O1-mediated downregulation of glucose-6-phosphatase[J].Mol Imag Biol,2017.
[10]Michalski CW,Hackert T,Büchler MW,et al.Targeting metabolism in pancreatic cancer[J].Lancet Oncol,2017,18(6):699-700.
[11]Bates SE.Pancreatic cancer:challenge and inspiration[J].Clin Cancer Res,2017,23(7):1628.
[12]Burk KS,Lo GC,Gee MS,et al.Imaging and screening of pancreatic cancer[J].Radiol Clin North Am,2017,55(6):1223-1234.
[13]Jason CH,Laura AM,Adam CM,et al.Novel patient-derived xenograft mouse model for pancreatic acinar cell carcinoma demonstrates single agent activity of oxaliplatin[J].J Translat Med,2016,14(1):129-143.
[14]Perez MR,Fleming JB.Patient-derived tumor xenograft models promise,potential and practice[M].New York:Academic Press,2016,49-82.
[15]Houghton JL,Zeglis BM,Abdelatti D,et al.Site-specifically labeled CA19-9-targeted immunoconjugates for the PET,NIRF,and multimodal PET/NIRF imaging of pancreatic cancer[J].Proc Nat Acad Sci U S A,2015,112(52):15850-15855.
[16]Khamis RY,Woollard KJ,Hyde GD,et al.Near infrared fluorescence(NIRF)molecular imaging of oxidized LDL with an autoantibody in experimental atherosclerosis[J].Sci Rep,2016,6:21785.
[17]Zhang CQ,Zhao Y,Zhang H,et al.The application of heptamethine cyanine dye DZ-1 and indocyanine green for imaging and targeting in xenograft models of hepatocellular carcinoma[J].Int J Mol Sci,2017,18(6):1332-1346.
[18]张彩勤,张海,赵勇,等.近红外荧光染料IR-783介导的肿瘤成像[J].中国实验动物学报,2014,22(2):17-20.Zhang CQ,Zhang H,Zhao Y,et al.Near infrared heptamethine cyanine dye IR-783-mediated tumor imaging[J].ACTA Lab Animalis Sci Sin,2014,22(2):17-20.
[19]Valtorta S,Moro M,Prisinzano G,et al.Metabolic evaluation of non-small cell lung cancer patient-derived xenografts models using 18F-FDG PET:potential tools for early therapy response[J].J Nucl Med,2017,58(1):42-47.
[20]Kiyuna T,Murakami T,Tome Y,et al.Labeling the stroma of a patient-derived orthotopic xenograft(PDOX)mouse model of undifferentiated pleomorphic soft-tissue sarcoma with red fluorescent protein for rapid non-invasive imaging for drug screening[J].J Cell Biochem,2017,118(2):361-365.
[21]Katherine BG,Rachel AA,Peter MV,et al.STR allele sequence variation:Current knowledge and future issues[J].Forensic Sci Int Genet,2015,18:118-130.
[22]Mohan B.Pathological diagnosis in pancreatic cancer preoperatively—a reality or myth?[J].Eur J Cancer,2017,72:79.
[23]Huang Z,Jiang Z,Zhao C,et al.Simple and effective label-free electrochemical immunoassay for carbohydrate antigen 19-9based on polythionine-Au composites as enhanced sensing signals for detecting different clinical samples[J].Int J Nanomed,2017,12:3049-3058.
[2]Smith RA,Andrews KS,Brooks D,et al.Cancer screening in the United States,2017:a review of current American Cancer society guidelines and current issues in cancer screening[J].CA Cancer J Clin,2017,67(2):100-121.
[3]Siegel RL,Miller KD,Jemal A.Cancer statistics,2017[J].CA Cancer J Clin,2017,67(1):7-30.
[4]Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.
[5]Rosfjord E,Lucas J,Li G,et al.Advances in patient-derived tumor xenografts:from target identification to predicting clinical response rates in oncology[J].Biochem Pharmacol,2014,91(2):135-43.
[6]Murakami T,Murata T,Kawaguchi K,et al.Cervical cancer patient-derived orthotopic xenograft(PDOX)is sensitive to cisplatinum and resistant to nab-paclitaxel[J].Anticancer Res,2017,37(1):61-65.
[7]Kawaguchi K,Igarashi K,Murakami T,et al.Tumor-targeting Salmonella typhimurium A1-R sensitizes melanoma with a BRAFV600E mutation to vemurafenib in a patient-derived orthotopic xenograft(PDOX)nude mouse model[J].J Cell Biochem,2017,118(8):2314-2319.
[8]Rapic S,Vangestel C,Verhaeghe J,et al.Characterization of an orthotopic colorectal cancer mouse model and its feasibility for accurate quantification in positron emission tomography[J].Mol Imaging Biol,2017,20:1536-1632.
[9]Wang Z,Kang F,Gao Y,et al.Metformin promotes 2-deoxy-2-[18F]fluoro-D-glucose uptake in hepatocellular carcinoma cells through Fox O1-mediated downregulation of glucose-6-phosphatase[J].Mol Imag Biol,2017.
[10]Michalski CW,Hackert T,Büchler MW,et al.Targeting metabolism in pancreatic cancer[J].Lancet Oncol,2017,18(6):699-700.
[11]Bates SE.Pancreatic cancer:challenge and inspiration[J].Clin Cancer Res,2017,23(7):1628.
[12]Burk KS,Lo GC,Gee MS,et al.Imaging and screening of pancreatic cancer[J].Radiol Clin North Am,2017,55(6):1223-1234.
[13]Jason CH,Laura AM,Adam CM,et al.Novel patient-derived xenograft mouse model for pancreatic acinar cell carcinoma demonstrates single agent activity of oxaliplatin[J].J Translat Med,2016,14(1):129-143.
[14]Perez MR,Fleming JB.Patient-derived tumor xenograft models promise,potential and practice[M].New York:Academic Press,2016,49-82.
[15]Houghton JL,Zeglis BM,Abdelatti D,et al.Site-specifically labeled CA19-9-targeted immunoconjugates for the PET,NIRF,and multimodal PET/NIRF imaging of pancreatic cancer[J].Proc Nat Acad Sci U S A,2015,112(52):15850-15855.
[16]Khamis RY,Woollard KJ,Hyde GD,et al.Near infrared fluorescence(NIRF)molecular imaging of oxidized LDL with an autoantibody in experimental atherosclerosis[J].Sci Rep,2016,6:21785.
[17]Zhang CQ,Zhao Y,Zhang H,et al.The application of heptamethine cyanine dye DZ-1 and indocyanine green for imaging and targeting in xenograft models of hepatocellular carcinoma[J].Int J Mol Sci,2017,18(6):1332-1346.
[18]张彩勤,张海,赵勇,等.近红外荧光染料IR-783介导的肿瘤成像[J].中国实验动物学报,2014,22(2):17-20.Zhang CQ,Zhang H,Zhao Y,et al.Near infrared heptamethine cyanine dye IR-783-mediated tumor imaging[J].ACTA Lab Animalis Sci Sin,2014,22(2):17-20.
[19]Valtorta S,Moro M,Prisinzano G,et al.Metabolic evaluation of non-small cell lung cancer patient-derived xenografts models using 18F-FDG PET:potential tools for early therapy response[J].J Nucl Med,2017,58(1):42-47.
[20]Kiyuna T,Murakami T,Tome Y,et al.Labeling the stroma of a patient-derived orthotopic xenograft(PDOX)mouse model of undifferentiated pleomorphic soft-tissue sarcoma with red fluorescent protein for rapid non-invasive imaging for drug screening[J].J Cell Biochem,2017,118(2):361-365.
[21]Katherine BG,Rachel AA,Peter MV,et al.STR allele sequence variation:Current knowledge and future issues[J].Forensic Sci Int Genet,2015,18:118-130.
[22]Mohan B.Pathological diagnosis in pancreatic cancer preoperatively—a reality or myth?[J].Eur J Cancer,2017,72:79.
[23]Huang Z,Jiang Z,Zhao C,et al.Simple and effective label-free electrochemical immunoassay for carbohydrate antigen 19-9based on polythionine-Au composites as enhanced sensing signals for detecting different clinical samples[J].Int J Nanomed,2017,12:3049-3058.