BACKGROUND Hepatocellular carcinoma(HCC)ranks second in terms of cancer mortality worldwide.Molecular magnetic resonance imaging(MRI)targeting HCC biomarkers such as alpha-fetoprotein(AFP)or glypican-3(GPC3)offers new...BACKGROUND Hepatocellular carcinoma(HCC)ranks second in terms of cancer mortality worldwide.Molecular magnetic resonance imaging(MRI)targeting HCC biomarkers such as alpha-fetoprotein(AFP)or glypican-3(GPC3)offers new strategies to enhance specificity and help early diagnosis of HCC.However,the existing iron oxide nanoparticle-based MR molecular probes singly target AFP or GPC3,which may hinder their efficiency to detect heterogeneous micro malignant HCC tumors<1 cm(MHCC).We hypothesized that the strategy of double antibody-conjugated iron oxide nanoparticles which simultaneously target AFP and GPC3 antigens may potentially be used to overcome the tumor heterogeneity and enhance the detection rate for MRI-based MHCC diagnosis.AIM To synthesize an AFP/GPC3 double antibody-labeled iron oxide MRI molecular probe and to assess its impact on MRI specificity and sensitivity at the cellular level.METHODS A double antigen-targeted MRI probe for MHCC anti-AFP-USPIO-anti-GPC3(UAG)was developed by simultaneously conjugating AFP andGPC3 antibodies to a 5 nm ultra-small superparamagnetic iron oxide nanoparticle(USPIO).At the same time,the singly labeled probes of anti-AFP-USPIO(UA)and anti-GPC3-USPIO(UG)and non-targeted USPIO(U)were also prepared for comparison.The physical characterization including morphology(transmission electron microscopy),hydrodynamic size,and zeta potential(dynamic light scattering)was conducted for each of the probes.The antigen targeting and MRI ability for these four kinds of USPIO probes were studied in the GPC3-expressing murine hepatoma cell line Hepa1-6/GPC3.First,AFP and GPC3 antigen expression in Hepa1-6/GPC3 cells was confirmed by flow cytometry and immunocytochemistry.Then,the cellular uptake of USPIO probes was investigated by Prussian blue staining assay and in vitro MRI(T2-weighted and T2-map)with a 3.0 Tesla clinical MR scanner.RESULTS Our data showed that the double antibody-conjugated probe UAG had the best specificity in targeting Hepa1-6/GPC3 cells expressing AFP and GPC3 antigens compared with single antibody-conjugated and unconjugated USPIO probes.The iron Prussian blue staining and quantitative T2-map MRI analysis showed that,compared with UA,UG,and U,the uptake of double antigen-targeted UAG probe demonstrated a 23.3%(vs UA),15.4%(vs UG),and 57.3%(vs U)increased Prussian stained cell percentage and a 14.93%(vs UA),9.38%(vs UG),and 15.3%(vs U)reduction of T2 relaxation time,respectively.Such bi-specific probe might have the potential to overcome tumor heterogeneity.Meanwhile,the coupling of two antibodies did not influence the magnetic performance of USPIO,and the relatively small hydrodynamic size(59.60±1.87 nm)of double antibodyconjugated USPIO probe makes it a viable candidate for use in MHCC MRI in vivo,as they are slowly phagocytosed by macrophages.CONCLUSION The bi-specific probe presents enhanced targeting efficiency and MRI sensitivity to HCC cells than singly-or non-targeted USPIO,paving the way for in vivo translation to further evaluate its clinical potential.展开更多
AIM: To investigate the anti-tumor effect and mechanisms of magnetic nanoparticles targeting hepatocellular carcinoma. METHODS: Human hepatocellular carcinoma was induced in nude mice, and the mice were randomly divid...AIM: To investigate the anti-tumor effect and mechanisms of magnetic nanoparticles targeting hepatocellular carcinoma. METHODS: Human hepatocellular carcinoma was induced in nude mice, and the mice were randomly divided into group A receiving normal saline, group B receiving magnetic nanoparticles containing 5-fluorouracil (5-FU), group C receiving 5-FU, and group D receiving magnetic nanoparticles containing 5-FU with a magnetic field built in tumor tissues. The tumor volume was measured on the day before treatment and 1, 4, 7, 10 and 13 d after treatment. Tumor tissues were isolated for examination of the expression of bcl-2, bax and caspase 3 by immunohistochemical method, reverse transcription polymerase chain reaction and Western blotting. RESULTS: The tumor volume was markedly lower in groups C and D than in groups A and B (group C or D vs group A or B, P < 0.01). The volume was markedly lower in group D than in group C (P < 0.05). The expression of protein and mRNA of bcl-2 was markedly lower in groups C and D than in groups A and B (group C or D vs group A or B, P < 0.01), and was markedly lower in group D than in group C (P < 0.01). The expression of bax and caspase 3 in groups C and D was signif icantly increased, compared with that in groups A and B (P < 0.01). CONCLUSION: The targeted magnetic nanoparticles containing 5-FU can improve the chemotherapeutic effect of 5-FU against hepatocellular carcinoma by decreasing the expression of bcl-2 gene, and increasing the expression of bax and caspase 3 genes.展开更多
Liver cancer is one of the most common cancers in the world. Of all types of livercancer, hepatocellular carcinoma (HCC) is known to be the most frequent primaryliver malignancy and has seriously compromised the healt...Liver cancer is one of the most common cancers in the world. Of all types of livercancer, hepatocellular carcinoma (HCC) is known to be the most frequent primaryliver malignancy and has seriously compromised the health status of the generalpopulation. Locoregional thermal ablation techniques such as radiofrequency andmicrowave ablation, have attracted attention in clinical practice as an alternativestrategy for HCC treatment. However, their aggressive thermal effect may causeundesirable complications such as hepatic decompensation, hemorrhage, bile ductinjury, extrahepatic organ injuries, and skin burn. In recent years, photodynamictherapy (PDT), a gentle locoregional treatment, has attracted attention in ablationtherapy for patients with superficial or luminal tumors as an alternative treatmentstrategy. However, some inherent defects and extrinsic factors of PDT havelimited its use in clinical practice for deep-seated HCC. In this contribution, theaim is to summarize the current status and challenges of PDT in HCC treatmentand provide potential strategies to overcome these deficiencies in further clinicaltranslational practice.展开更多
Currently, sorafenib is the only systemic therapy capable of increasing overall survival of hepatocellular carcinoma patients. Unfortunately, its side effects, particularly its overall toxicity, limit the therapeutic ...Currently, sorafenib is the only systemic therapy capable of increasing overall survival of hepatocellular carcinoma patients. Unfortunately, its side effects, particularly its overall toxicity, limit the therapeutic response that can be achieved. Superparamagnetic iron oxide nanoparticles （SPIONs） are very attractive for drug delivery because they can be targeted to specific sites in the body through application of a magnetic field, thus improving intratumoral accumulation and reducing adverse effects. Here, nanoformulations based on polyethylene glycol modified phospholipid micelles, loaded with both SPIONs and sorafenib, were successfully prepared and thoroughly investigated by complementary techniques. This nanovector system provided effective drug delivery, had an average hydrodynamic diameter of about 125 nm, had good stability in aqueous medium, and allowed controlled drug loading. Magnetic analysis allowed accurate determination of the amount of SPIONs embedded in each micelle. An in vitro system was designed to test whether the SPION micelles can be efficiently held using a magnetic field under typical flow conditions found in the human liver. Human hepatocellular carcinoma （HepG2） cells were selected as an in vitro system to evaluate tumor cell targeting efficacy of the superparamagnetic micelles loaded with sorafenib. These experiments demonstrated that this delivery platform is able to enhance sorafenib＇s antitumor effectiveness by magnetic targeting. The magnetic nanovectors described here represent promising candidates for targeting specific hepatic tumor sites, where selective release of sorafenib can improve its efficacy and safety profile.展开更多
Despite being a common therapy for hepatocellular carcinoma(HCC),insufficient thermal ablation can leave behind tumor residues that can cause recurrence.This is believed to augment M2 inflammatory macrophages that usu...Despite being a common therapy for hepatocellular carcinoma(HCC),insufficient thermal ablation can leave behind tumor residues that can cause recurrence.This is believed to augment M2 inflammatory macrophages that usually play a pro-tumorigenic role.To address this problem,we designed D-mannose-chelated iron oxide nanoparticles(man-IONPs) to polarize M2-like macrophages into the antitumor Ml phenotype.In vitro and in vivo experiments demonstrated that man-IONPs specifically targeted M2-like macrophages and accumulated in peri-ablation zones after macrophage infiltration was augmented under insufficient microwave ablation(MWA).The nanoparticles simultaneously induced polarization of pro-tumorigenic M2 macrophages into antitumor M1 phenotypes,enabling the transformation of the immunosuppressive microenvironment into an immunoactivating one.Post-MWA macrophage polarization exerted robust inhibitory effects on HCC progression in a well-established orthotopic liver cancer mouse model.Thus,combining thermal ablation with man-IONPs can salvage residual tumors after insufficient MWA.These results have strong potential for clinical translation.展开更多
目的观察血管内皮生长因子C(VEGF-C)抗体与超顺磁性氧化铁颗粒(USPIO)连接的靶向分子探针(VEGF-C-USPIO)在大鼠肝细胞癌(HCC)模型体内的MR成像特点,并探讨其临床意义。方法采用诱导法建立大鼠原位肝癌模型,将30只SD大鼠随机分为实验组(n...目的观察血管内皮生长因子C(VEGF-C)抗体与超顺磁性氧化铁颗粒(USPIO)连接的靶向分子探针(VEGF-C-USPIO)在大鼠肝细胞癌(HCC)模型体内的MR成像特点,并探讨其临床意义。方法采用诱导法建立大鼠原位肝癌模型,将30只SD大鼠随机分为实验组(n=20)和对照组(n=10)。分别于鼠尾静脉注射靶向探针VEGF-C-USPIO和非靶向探针USPIO,并于注射前及注射后1 h对大鼠行MR扫描成像,测量其肝脏肿瘤与周围肝组织的T2WI信号强度,计算噪声比(CNR),比较增强前后2组之间CNR的差异。扫描结束后取动物肝脏进行HE染色明确大鼠肝癌病理类型;普鲁士蓝染色验证肿瘤组织细胞中铁含量;免疫组化染色验证肝癌组织中VEGF-C的表达情况。实验组与对照组间的比较采用独立样本t检验,实验组或对照组内注射对比剂前后的比较采用配对样本t检验。结果 30只大鼠全部诱癌成功,病理学诊断为HCC,成瘤率100%。实验组注射靶向对比剂VEGF-C-USPIO后1 h与注射前CNR比较,差异有统计学意义(2.11±0.23 vs 3.47±0.45,t=-13.15,P<0.001);对照组注射非靶向对比剂USPIO后1 h与注射前CNR之间差异无统计学意义(3.51±0.14 vs 3.82±0.61,t=-1.40,P=0.192);2组大鼠注射对比剂后的CNR比较,差异有统计学意义(t=17.60,P<0.001)。对大鼠肝脏标本行HE染色,结果证实为HCC;免疫组化染色显示,VEGF-C主要在肝癌细胞胞膜及胞浆中表达;普鲁士蓝染色显示,实验组肿瘤组织内蓝染铁颗粒较对照组明显增多。结论所合成的分子靶向探针VEGF-C-USPIO对大鼠HCC模型具有较好的主动靶向作用,能够通过MR信号强度的变化实现HCC的特异性成像,为HCC的早期诊断提供影像学依据。展开更多
基金Supported by CAMS Innovation Fund for Medical Sciences,No.2016-I2M-1-001PUMC Youth Fund,No.2017320010+1 种基金Chinese Academy of Medical Sciences Research Fund,No.ZZ2016B01Beijing HopeRun Special Fund of Cancer Foundation of China,No.LC2016B15
文摘BACKGROUND Hepatocellular carcinoma(HCC)ranks second in terms of cancer mortality worldwide.Molecular magnetic resonance imaging(MRI)targeting HCC biomarkers such as alpha-fetoprotein(AFP)or glypican-3(GPC3)offers new strategies to enhance specificity and help early diagnosis of HCC.However,the existing iron oxide nanoparticle-based MR molecular probes singly target AFP or GPC3,which may hinder their efficiency to detect heterogeneous micro malignant HCC tumors<1 cm(MHCC).We hypothesized that the strategy of double antibody-conjugated iron oxide nanoparticles which simultaneously target AFP and GPC3 antigens may potentially be used to overcome the tumor heterogeneity and enhance the detection rate for MRI-based MHCC diagnosis.AIM To synthesize an AFP/GPC3 double antibody-labeled iron oxide MRI molecular probe and to assess its impact on MRI specificity and sensitivity at the cellular level.METHODS A double antigen-targeted MRI probe for MHCC anti-AFP-USPIO-anti-GPC3(UAG)was developed by simultaneously conjugating AFP andGPC3 antibodies to a 5 nm ultra-small superparamagnetic iron oxide nanoparticle(USPIO).At the same time,the singly labeled probes of anti-AFP-USPIO(UA)and anti-GPC3-USPIO(UG)and non-targeted USPIO(U)were also prepared for comparison.The physical characterization including morphology(transmission electron microscopy),hydrodynamic size,and zeta potential(dynamic light scattering)was conducted for each of the probes.The antigen targeting and MRI ability for these four kinds of USPIO probes were studied in the GPC3-expressing murine hepatoma cell line Hepa1-6/GPC3.First,AFP and GPC3 antigen expression in Hepa1-6/GPC3 cells was confirmed by flow cytometry and immunocytochemistry.Then,the cellular uptake of USPIO probes was investigated by Prussian blue staining assay and in vitro MRI(T2-weighted and T2-map)with a 3.0 Tesla clinical MR scanner.RESULTS Our data showed that the double antibody-conjugated probe UAG had the best specificity in targeting Hepa1-6/GPC3 cells expressing AFP and GPC3 antigens compared with single antibody-conjugated and unconjugated USPIO probes.The iron Prussian blue staining and quantitative T2-map MRI analysis showed that,compared with UA,UG,and U,the uptake of double antigen-targeted UAG probe demonstrated a 23.3%(vs UA),15.4%(vs UG),and 57.3%(vs U)increased Prussian stained cell percentage and a 14.93%(vs UA),9.38%(vs UG),and 15.3%(vs U)reduction of T2 relaxation time,respectively.Such bi-specific probe might have the potential to overcome tumor heterogeneity.Meanwhile,the coupling of two antibodies did not influence the magnetic performance of USPIO,and the relatively small hydrodynamic size(59.60±1.87 nm)of double antibodyconjugated USPIO probe makes it a viable candidate for use in MHCC MRI in vivo,as they are slowly phagocytosed by macrophages.CONCLUSION The bi-specific probe presents enhanced targeting efficiency and MRI sensitivity to HCC cells than singly-or non-targeted USPIO,paving the way for in vivo translation to further evaluate its clinical potential.
基金Supported by the Hi-Tech Research and Development Program of China, NO.2002AA214061
文摘AIM: To investigate the anti-tumor effect and mechanisms of magnetic nanoparticles targeting hepatocellular carcinoma. METHODS: Human hepatocellular carcinoma was induced in nude mice, and the mice were randomly divided into group A receiving normal saline, group B receiving magnetic nanoparticles containing 5-fluorouracil (5-FU), group C receiving 5-FU, and group D receiving magnetic nanoparticles containing 5-FU with a magnetic field built in tumor tissues. The tumor volume was measured on the day before treatment and 1, 4, 7, 10 and 13 d after treatment. Tumor tissues were isolated for examination of the expression of bcl-2, bax and caspase 3 by immunohistochemical method, reverse transcription polymerase chain reaction and Western blotting. RESULTS: The tumor volume was markedly lower in groups C and D than in groups A and B (group C or D vs group A or B, P < 0.01). The volume was markedly lower in group D than in group C (P < 0.05). The expression of protein and mRNA of bcl-2 was markedly lower in groups C and D than in groups A and B (group C or D vs group A or B, P < 0.01), and was markedly lower in group D than in group C (P < 0.01). The expression of bax and caspase 3 in groups C and D was signif icantly increased, compared with that in groups A and B (P < 0.01). CONCLUSION: The targeted magnetic nanoparticles containing 5-FU can improve the chemotherapeutic effect of 5-FU against hepatocellular carcinoma by decreasing the expression of bcl-2 gene, and increasing the expression of bax and caspase 3 genes.
基金the Wuhan Municipal Health Commission,No.WX14B22the National Natural Science Foundation of China,No.81874208 and No.81700425.
文摘Liver cancer is one of the most common cancers in the world. Of all types of livercancer, hepatocellular carcinoma (HCC) is known to be the most frequent primaryliver malignancy and has seriously compromised the health status of the generalpopulation. Locoregional thermal ablation techniques such as radiofrequency andmicrowave ablation, have attracted attention in clinical practice as an alternativestrategy for HCC treatment. However, their aggressive thermal effect may causeundesirable complications such as hepatic decompensation, hemorrhage, bile ductinjury, extrahepatic organ injuries, and skin burn. In recent years, photodynamictherapy (PDT), a gentle locoregional treatment, has attracted attention in ablationtherapy for patients with superficial or luminal tumors as an alternative treatmentstrategy. However, some inherent defects and extrinsic factors of PDT havelimited its use in clinical practice for deep-seated HCC. In this contribution, theaim is to summarize the current status and challenges of PDT in HCC treatmentand provide potential strategies to overcome these deficiencies in further clinicaltranslational practice.
文摘Currently, sorafenib is the only systemic therapy capable of increasing overall survival of hepatocellular carcinoma patients. Unfortunately, its side effects, particularly its overall toxicity, limit the therapeutic response that can be achieved. Superparamagnetic iron oxide nanoparticles （SPIONs） are very attractive for drug delivery because they can be targeted to specific sites in the body through application of a magnetic field, thus improving intratumoral accumulation and reducing adverse effects. Here, nanoformulations based on polyethylene glycol modified phospholipid micelles, loaded with both SPIONs and sorafenib, were successfully prepared and thoroughly investigated by complementary techniques. This nanovector system provided effective drug delivery, had an average hydrodynamic diameter of about 125 nm, had good stability in aqueous medium, and allowed controlled drug loading. Magnetic analysis allowed accurate determination of the amount of SPIONs embedded in each micelle. An in vitro system was designed to test whether the SPION micelles can be efficiently held using a magnetic field under typical flow conditions found in the human liver. Human hepatocellular carcinoma （HepG2） cells were selected as an in vitro system to evaluate tumor cell targeting efficacy of the superparamagnetic micelles loaded with sorafenib. These experiments demonstrated that this delivery platform is able to enhance sorafenib＇s antitumor effectiveness by magnetic targeting. The magnetic nanovectors described here represent promising candidates for targeting specific hepatic tumor sites, where selective release of sorafenib can improve its efficacy and safety profile.
基金supported by the National Natural Science Foundation of China (Grant Nos. 91859201, 92159305, 81971625, and 82030047)
文摘Despite being a common therapy for hepatocellular carcinoma(HCC),insufficient thermal ablation can leave behind tumor residues that can cause recurrence.This is believed to augment M2 inflammatory macrophages that usually play a pro-tumorigenic role.To address this problem,we designed D-mannose-chelated iron oxide nanoparticles(man-IONPs) to polarize M2-like macrophages into the antitumor Ml phenotype.In vitro and in vivo experiments demonstrated that man-IONPs specifically targeted M2-like macrophages and accumulated in peri-ablation zones after macrophage infiltration was augmented under insufficient microwave ablation(MWA).The nanoparticles simultaneously induced polarization of pro-tumorigenic M2 macrophages into antitumor M1 phenotypes,enabling the transformation of the immunosuppressive microenvironment into an immunoactivating one.Post-MWA macrophage polarization exerted robust inhibitory effects on HCC progression in a well-established orthotopic liver cancer mouse model.Thus,combining thermal ablation with man-IONPs can salvage residual tumors after insufficient MWA.These results have strong potential for clinical translation.
文摘目的观察血管内皮生长因子C(VEGF-C)抗体与超顺磁性氧化铁颗粒(USPIO)连接的靶向分子探针(VEGF-C-USPIO)在大鼠肝细胞癌(HCC)模型体内的MR成像特点,并探讨其临床意义。方法采用诱导法建立大鼠原位肝癌模型,将30只SD大鼠随机分为实验组(n=20)和对照组(n=10)。分别于鼠尾静脉注射靶向探针VEGF-C-USPIO和非靶向探针USPIO,并于注射前及注射后1 h对大鼠行MR扫描成像,测量其肝脏肿瘤与周围肝组织的T2WI信号强度,计算噪声比(CNR),比较增强前后2组之间CNR的差异。扫描结束后取动物肝脏进行HE染色明确大鼠肝癌病理类型;普鲁士蓝染色验证肿瘤组织细胞中铁含量;免疫组化染色验证肝癌组织中VEGF-C的表达情况。实验组与对照组间的比较采用独立样本t检验,实验组或对照组内注射对比剂前后的比较采用配对样本t检验。结果 30只大鼠全部诱癌成功,病理学诊断为HCC,成瘤率100%。实验组注射靶向对比剂VEGF-C-USPIO后1 h与注射前CNR比较,差异有统计学意义(2.11±0.23 vs 3.47±0.45,t=-13.15,P<0.001);对照组注射非靶向对比剂USPIO后1 h与注射前CNR之间差异无统计学意义(3.51±0.14 vs 3.82±0.61,t=-1.40,P=0.192);2组大鼠注射对比剂后的CNR比较,差异有统计学意义(t=17.60,P<0.001)。对大鼠肝脏标本行HE染色,结果证实为HCC;免疫组化染色显示,VEGF-C主要在肝癌细胞胞膜及胞浆中表达;普鲁士蓝染色显示,实验组肿瘤组织内蓝染铁颗粒较对照组明显增多。结论所合成的分子靶向探针VEGF-C-USPIO对大鼠HCC模型具有较好的主动靶向作用,能够通过MR信号强度的变化实现HCC的特异性成像,为HCC的早期诊断提供影像学依据。
