NIRF(Np95/ICBP90-like RING finger protein)是2002年发现的一种核蛋白,其功能涉及细胞增殖调节、蛋白多聚泛素化降解、细胞癌变进程控制等领域.已有研究报道,NIRF能与p53相互作用,NIRF本身也是一个高度调节蛋白,在细胞正常的生理状态...NIRF(Np95/ICBP90-like RING finger protein)是2002年发现的一种核蛋白,其功能涉及细胞增殖调节、蛋白多聚泛素化降解、细胞癌变进程控制等领域.已有研究报道,NIRF能与p53相互作用,NIRF本身也是一个高度调节蛋白,在细胞正常的生理状态下发挥泛素化E3连接酶的作用,结合p53并将其降解,但NIRF与p53结合的蛋白结合域目前尚不清楚.本文研究证明,NIRF能与p53结合成复合体参与泛素化蛋白降解途径,并测定出NIRF与p53结合的区域.为了检测NIRF的蛋白结合域,将空载体和NIRF缺失突变体质粒分别转染于HEK293细胞,蛋白表达水平通过Western印迹用两种抗体分别检测.结果显示,所有的突变体都能在细胞中表达,并且两种抗体检测结果完全一致.同时,免疫共沉淀技术用于进一步分析实验结果.由于泛素化蛋白通常伴随蛋白酶体通路介导的降解,免疫共沉淀的蛋白纯化过程中用蛋白酶体抑制剂MG-132以抑制蛋白降解.本研究结果显示,NIRF通过PHD区域与p53形成复合体.该复合体可能参与蛋白分选、蛋白降解、DNA修复以及细胞凋亡等一系列重要的细胞活动,从而形成与细胞增殖相关的新的信号通路,在肿瘤的发生发展中可能发挥某种程度的作用.展开更多
为研究NIRF(Np95/ICBP-90 like RING finger protein)对乙型肝炎病毒(hepatitis B virus,HBV)的复制以及与乙型肝炎病毒共价环状闭合DNA(HBV cccDNA)结合的组蛋白H3乙酰化的影响,采用脂质体转染将pGEM-HBV1.3+pFLAG、pGEM-HBV1.3+pFLAG-...为研究NIRF(Np95/ICBP-90 like RING finger protein)对乙型肝炎病毒(hepatitis B virus,HBV)的复制以及与乙型肝炎病毒共价环状闭合DNA(HBV cccDNA)结合的组蛋白H3乙酰化的影响,采用脂质体转染将pGEM-HBV1.3+pFLAG、pGEM-HBV1.3+pFLAG-NIRF、pGEM-HBV1.3质粒分别转入HepG2细胞,Western blot检测NIRF蛋白的表达,用ELISA结合RT-PCR检测HBsAg、HBeAg以及HBV cccDNA的量并同时说明HBV在细胞内是完成完整复制表达的,采用染色质免疫共沉淀(ChIP)的方法检测与HBV cccDNA结合的组蛋白H3以及H3乙酰化水平的动态变化。结果显示,NIRF蛋白下调HBV标志物HBeAg、HBsAg的分泌以及HBV cccDNA的表达,表明其对HBV复制具有抑制作用;组蛋白H3及乙酰化的组蛋白H3都与HBV cccDNA的动态变化水平呈现相似的平行性,而NIRF蛋白也明显抑制组蛋白H3的表达水平和乙酰化水平。结论证实NIRF不仅能抑制HBV在肝癌细胞中的复制,而且能下调与HBV cccDNA结合的组蛋白H3和乙酰化组蛋白H3的表达。期待NIRF能为后续的HBV致病机理、HBV复制表观遗传学水平研究及有效抗病毒药物的研究与开发提供理论上的支持与帮助。展开更多
随着对NIRF(Np95/ICBP-90 like RING finger protein)研究的深入,其功能已涉及细胞癌变进程以及表观遗传学等领域.近期研究显示,NIRF能与HBc(hepatitis B virus core protein)相互结合,但其对乙型肝炎病毒(HBV)抗原表达的影响尚不明确....随着对NIRF(Np95/ICBP-90 like RING finger protein)研究的深入,其功能已涉及细胞癌变进程以及表观遗传学等领域.近期研究显示,NIRF能与HBc(hepatitis B virus core protein)相互结合,但其对乙型肝炎病毒(HBV)抗原表达的影响尚不明确.本文通过转染pAAV-HBV1.3质粒和高压水动力法尾静脉注射BALB/C小鼠,建立乙型肝炎病毒的细胞和动物模型,研究NIRF对乙型肝炎病毒抗原表达的影响.ELISA检测细胞上清和小鼠血清中HBsAg、HBeAg的分泌和表达情况,Western印迹或免疫组化染色技术检测HBcAg.结果显示,乙型肝炎病毒抗原分泌的细胞以及小动物模型建立成功,并且无论在体内外,NIRF都能对它们的表达起抑制作用,期待能为后续的HBV致病机理以及治疗药物的研究提供支持与帮助.展开更多
目前对NIRF(Np95/ICBP-90 like RING finger protein)的研究正朝着细胞癌变进程以及表观遗传学的方向发展,但在体内NIRF对乙型肝炎病毒(HBV)的复制及表达的影响,目前尚不明确.通过高压水动力法转染HBV小鼠模型,不同时间点收集血液和肝...目前对NIRF(Np95/ICBP-90 like RING finger protein)的研究正朝着细胞癌变进程以及表观遗传学的方向发展,但在体内NIRF对乙型肝炎病毒(HBV)的复制及表达的影响,目前尚不明确.通过高压水动力法转染HBV小鼠模型,不同时间点收集血液和肝组织标本,荧光定量PCR检测血清及肝组织中病毒载量,WesternBlot检测肝组织HBc(hepatitis B virus core protein)表达,ELISA检测血清HBeAg表达,并通过免疫组化染色检测HBsAg、HBcAg在肝组织中的定位及表达.小鼠转染pAAV-HBV1.3和NIRF以后,血清及肝组织病毒载量降低(n=3,P<0.05),HBc蛋白及HBV相关抗原的表达受到抑制,说明在水动力法转染HBV小鼠模型中NIRF对HBV的复制及表达起到抑制作用,期待能为后续的HBV致病机理及治疗药物的研究与开发提供支持与帮助.展开更多
Intelligent nanoplatform that combines multimodal imaging and therapeutic effects holds great promise for precise and efficient cancer therapy.Herein,folate-targeted polymersomes with stimuli-responsiveness were fabri...Intelligent nanoplatform that combines multimodal imaging and therapeutic effects holds great promise for precise and efficient cancer therapy.Herein,folate-targeted polymersomes with stimuli-responsiveness were fabricated and evaluated by near-infrared fluorescence(NIRF)and optical coherence tomography angiography(OCTA)dual-imaging for photo-chemo-antiangiogenic therapy against cancer.The folate-targeted polymersomes(FA-MIT-SIPS)not only integrated ammonium bicarbonate(ABC)and mitoxantrone(MIT)into their hydrophilic cavity but also encapsulated indocyanine green(ICG)and sorafenib(SOR)within their hydrophobic layer.NIRF imaging demonstrated that FA-MIT-SIPS effectively accumulated and retained in the tumors.Upon 808 nm laser irradiation,the ICG produced hyperthermia and reactive oxygen species(ROS)for efficient photothermal and photodynamic therapy.In addition,the decomposition of ABC in responsive to acidic tumor environment and ICG-induced hyperthermia accelerated drug release.The released MIT accumulated in nucleus to inhibit DNA synthesis,while the released SOR destructed tumor vascularization.Notably,OCTA imaging was applied to observe the tumor blood flow upon the combination therapy,demonstrating that FA-MIT-SIPS obviously decreased the vessels area density.Moreover,the synergistic photo-chemo-antiangiogenic therapy of FA-MIT-SIPS achieved excellent antitumor effect with 40%of the 4T1 tumor-bearing mice being completely cured without recurrence.The multifunctional polymersomes provide a promising dual-modal imaging-evaluated synergistic strategy for tumor therapy.展开更多
Cervical cancer,the most common gynecological malignancy,significantly and adversely af-fects women’s physical health and well-being.Traditional surgical interventions and chemotherapy,while potentially effective,oft...Cervical cancer,the most common gynecological malignancy,significantly and adversely af-fects women’s physical health and well-being.Traditional surgical interventions and chemotherapy,while potentially effective,often entail serious side effects that have led to an urgent need for novel therapeutic methods.Photothermal therapy(PTT)has emerged as a promising approach due to its ability to minimize damage to healthy tissue.Connecting a biothiol detection group to PTT-sensitive molecules can improve tumor targeting and further minimize potential side effects.In this study,we developed a near-infrared fluorescence(NIRF)/photoacoustic(PA)dual-mode probe,S-NBD,which demonstrated robust PTT per-formance.This innovative probe is capable of activating NIRF/PA signals to enable the detection of bio-thiols with high emission wavelength(838 nm)and large Stokes shift(178 nm),allowing for in vivo monitoring of cancer cells.Additionally,the probe achieved an outstanding photothermal conversion ef-ficiency of 67.1%.The application of laser irradiation(660 nm,1.0 W/cm^(2),5 min)was able to achieve complete tumor ablation without recurrence.In summary,this seminal study presents a pioneering NIRF/PA dual-mode dicyanoisophorone-based probe for biothiol imaging,incorporating features from PTT for the first time.This pioneering approach achieves the dual objectives of improving tumor diagnosis and treatment.展开更多
文摘NIRF(Np95/ICBP90-like RING finger protein)是2002年发现的一种核蛋白,其功能涉及细胞增殖调节、蛋白多聚泛素化降解、细胞癌变进程控制等领域.已有研究报道,NIRF能与p53相互作用,NIRF本身也是一个高度调节蛋白,在细胞正常的生理状态下发挥泛素化E3连接酶的作用,结合p53并将其降解,但NIRF与p53结合的蛋白结合域目前尚不清楚.本文研究证明,NIRF能与p53结合成复合体参与泛素化蛋白降解途径,并测定出NIRF与p53结合的区域.为了检测NIRF的蛋白结合域,将空载体和NIRF缺失突变体质粒分别转染于HEK293细胞,蛋白表达水平通过Western印迹用两种抗体分别检测.结果显示,所有的突变体都能在细胞中表达,并且两种抗体检测结果完全一致.同时,免疫共沉淀技术用于进一步分析实验结果.由于泛素化蛋白通常伴随蛋白酶体通路介导的降解,免疫共沉淀的蛋白纯化过程中用蛋白酶体抑制剂MG-132以抑制蛋白降解.本研究结果显示,NIRF通过PHD区域与p53形成复合体.该复合体可能参与蛋白分选、蛋白降解、DNA修复以及细胞凋亡等一系列重要的细胞活动,从而形成与细胞增殖相关的新的信号通路,在肿瘤的发生发展中可能发挥某种程度的作用.
文摘为研究NIRF(Np95/ICBP-90 like RING finger protein)对乙型肝炎病毒(hepatitis B virus,HBV)的复制以及与乙型肝炎病毒共价环状闭合DNA(HBV cccDNA)结合的组蛋白H3乙酰化的影响,采用脂质体转染将pGEM-HBV1.3+pFLAG、pGEM-HBV1.3+pFLAG-NIRF、pGEM-HBV1.3质粒分别转入HepG2细胞,Western blot检测NIRF蛋白的表达,用ELISA结合RT-PCR检测HBsAg、HBeAg以及HBV cccDNA的量并同时说明HBV在细胞内是完成完整复制表达的,采用染色质免疫共沉淀(ChIP)的方法检测与HBV cccDNA结合的组蛋白H3以及H3乙酰化水平的动态变化。结果显示,NIRF蛋白下调HBV标志物HBeAg、HBsAg的分泌以及HBV cccDNA的表达,表明其对HBV复制具有抑制作用;组蛋白H3及乙酰化的组蛋白H3都与HBV cccDNA的动态变化水平呈现相似的平行性,而NIRF蛋白也明显抑制组蛋白H3的表达水平和乙酰化水平。结论证实NIRF不仅能抑制HBV在肝癌细胞中的复制,而且能下调与HBV cccDNA结合的组蛋白H3和乙酰化组蛋白H3的表达。期待NIRF能为后续的HBV致病机理、HBV复制表观遗传学水平研究及有效抗病毒药物的研究与开发提供理论上的支持与帮助。
文摘随着对NIRF(Np95/ICBP-90 like RING finger protein)研究的深入,其功能已涉及细胞癌变进程以及表观遗传学等领域.近期研究显示,NIRF能与HBc(hepatitis B virus core protein)相互结合,但其对乙型肝炎病毒(HBV)抗原表达的影响尚不明确.本文通过转染pAAV-HBV1.3质粒和高压水动力法尾静脉注射BALB/C小鼠,建立乙型肝炎病毒的细胞和动物模型,研究NIRF对乙型肝炎病毒抗原表达的影响.ELISA检测细胞上清和小鼠血清中HBsAg、HBeAg的分泌和表达情况,Western印迹或免疫组化染色技术检测HBcAg.结果显示,乙型肝炎病毒抗原分泌的细胞以及小动物模型建立成功,并且无论在体内外,NIRF都能对它们的表达起抑制作用,期待能为后续的HBV致病机理以及治疗药物的研究提供支持与帮助.
文摘目前对NIRF(Np95/ICBP-90 like RING finger protein)的研究正朝着细胞癌变进程以及表观遗传学的方向发展,但在体内NIRF对乙型肝炎病毒(HBV)的复制及表达的影响,目前尚不明确.通过高压水动力法转染HBV小鼠模型,不同时间点收集血液和肝组织标本,荧光定量PCR检测血清及肝组织中病毒载量,WesternBlot检测肝组织HBc(hepatitis B virus core protein)表达,ELISA检测血清HBeAg表达,并通过免疫组化染色检测HBsAg、HBcAg在肝组织中的定位及表达.小鼠转染pAAV-HBV1.3和NIRF以后,血清及肝组织病毒载量降低(n=3,P<0.05),HBc蛋白及HBV相关抗原的表达受到抑制,说明在水动力法转染HBV小鼠模型中NIRF对HBV的复制及表达起到抑制作用,期待能为后续的HBV致病机理及治疗药物的研究与开发提供支持与帮助.
基金supported by the National Natural Science Foundation of China(Nos.82072059 and 82172090)the Fundamental Research Funds for the Central Universities(No.2019PT320028)+2 种基金Tianjin Municipal Natural Science Foundation(No.20JCYBJC00030)CAMS Initiative for Innovative Medicine(No.2021-I2M-1-058)Science and Technology Planning Project of Tianjin(No.18ZXSGSY00050).
文摘Intelligent nanoplatform that combines multimodal imaging and therapeutic effects holds great promise for precise and efficient cancer therapy.Herein,folate-targeted polymersomes with stimuli-responsiveness were fabricated and evaluated by near-infrared fluorescence(NIRF)and optical coherence tomography angiography(OCTA)dual-imaging for photo-chemo-antiangiogenic therapy against cancer.The folate-targeted polymersomes(FA-MIT-SIPS)not only integrated ammonium bicarbonate(ABC)and mitoxantrone(MIT)into their hydrophilic cavity but also encapsulated indocyanine green(ICG)and sorafenib(SOR)within their hydrophobic layer.NIRF imaging demonstrated that FA-MIT-SIPS effectively accumulated and retained in the tumors.Upon 808 nm laser irradiation,the ICG produced hyperthermia and reactive oxygen species(ROS)for efficient photothermal and photodynamic therapy.In addition,the decomposition of ABC in responsive to acidic tumor environment and ICG-induced hyperthermia accelerated drug release.The released MIT accumulated in nucleus to inhibit DNA synthesis,while the released SOR destructed tumor vascularization.Notably,OCTA imaging was applied to observe the tumor blood flow upon the combination therapy,demonstrating that FA-MIT-SIPS obviously decreased the vessels area density.Moreover,the synergistic photo-chemo-antiangiogenic therapy of FA-MIT-SIPS achieved excellent antitumor effect with 40%of the 4T1 tumor-bearing mice being completely cured without recurrence.The multifunctional polymersomes provide a promising dual-modal imaging-evaluated synergistic strategy for tumor therapy.
基金supported by Foundation of Ministry of Science and Technology of China(2022YFC2304203)National Natural Science Foundation of China(Nos.82073689,82273762 and 81973183)+1 种基金National Natural Science Foundation of Guangdong Province(No.2024A1515010642,China)Science and Technology Program of Guangzhou(No.201904010380,China).
文摘Cervical cancer,the most common gynecological malignancy,significantly and adversely af-fects women’s physical health and well-being.Traditional surgical interventions and chemotherapy,while potentially effective,often entail serious side effects that have led to an urgent need for novel therapeutic methods.Photothermal therapy(PTT)has emerged as a promising approach due to its ability to minimize damage to healthy tissue.Connecting a biothiol detection group to PTT-sensitive molecules can improve tumor targeting and further minimize potential side effects.In this study,we developed a near-infrared fluorescence(NIRF)/photoacoustic(PA)dual-mode probe,S-NBD,which demonstrated robust PTT per-formance.This innovative probe is capable of activating NIRF/PA signals to enable the detection of bio-thiols with high emission wavelength(838 nm)and large Stokes shift(178 nm),allowing for in vivo monitoring of cancer cells.Additionally,the probe achieved an outstanding photothermal conversion ef-ficiency of 67.1%.The application of laser irradiation(660 nm,1.0 W/cm^(2),5 min)was able to achieve complete tumor ablation without recurrence.In summary,this seminal study presents a pioneering NIRF/PA dual-mode dicyanoisophorone-based probe for biothiol imaging,incorporating features from PTT for the first time.This pioneering approach achieves the dual objectives of improving tumor diagnosis and treatment.
