用红外光谱研究胃癌细胞SGC7901光敏损伤后蛋白质的二级结构

本文用傅立叶变换红外光谱研究光动力作用胃癌细胞SGC7901后,蛋白质二级结构的变化。对蛋白质的酰胺I带,采用去卷积,二阶导数谱进行曲线拟合,得到蛋白质二级结构的峰个数和位置以及各二级结构的含量,结果显示蛋白质无规卷曲和β转角的含量上升,α螺旋、β折叠含量减少,蛋白质空间构象改变。实验结果表明,光动力对胃癌细胞SGC7901蛋白质二级结构造成明显损伤。     

利用拉曼光谱研究光动力作用对C6胶质瘤细胞的杀伤效应

在光敏剂浓度和光剂量一定的条件下,利用拉曼光谱技术研究光敏剂不同孵育时间对C6胶质瘤细胞光动力杀伤效应。实验分5个实验组(光敏剂孵育时间1 h,3 h,6 h,9 h和1个对照组),采集5组C6胶质瘤细胞的拉曼光谱。结果显示:与核酸和蛋白质相关的谱线,峰强和峰位都有很大的变化,孵育时间为1 h的谱线,各峰强度变化较小;孵育时间为6 h的谱线,各峰强度变化最大;孵育时间为9 h的谱线,各峰强度变化进入平台期。实验结果表明:在一定的时间范围内,光动力作用对C6胶质瘤细胞光动力杀伤效应,与光敏剂孵育时间成正相关     

用光谱法研究光敏剂血卟啉单甲醚对肿瘤细胞光动力作用的分子机制

测定了光敏剂血卟啉单甲醚对人宫颈癌细胞HeLa光敏作用后的傅里叶红外光谱。结果显示:光敏作用后,HeLa细胞磷酸二酯基团的对称伸缩振动峰1085cm^-1和不对称伸缩振动峰1246cm^-1蓝移,强度下降:蛋白质酰胺Ⅰ带1656cm^-1发生蓝移,酰胺Ⅱ带1546cm^-1出现红移;CH₂对称伸缩振动峰2858cm^-1,峰位蓝移2cm^-1,峰值明显减弱:细胞的蛋白质和核酸谱峰面积比值D1085/D1546降低。提示细胞中的DNA、蛋白质和磷脂结构受到损伤。结果表明:DNA、蛋白质和磷脂是血卟啉单甲醚光敏作用的主要靶分子。     

光动力疗法中最佳光动力治疗剂量的研究

为了研究光动力过程中光剂量和光敏剂剂量对光动力损伤效果的影响,基于肿瘤组织中光动力损伤剂量的数学模型以及光动力损伤剂量与组织中光剂量、光敏剂剂量、氧浓度之间的函数关系,用数学方法模拟研究了给定模型组织中光及光敏剂的有效吸收剂量与氧浓度之间的关系,并用ALA-PDT实验研究了k562细胞悬浮液中不同药物剂量对光动力损伤效果的影响.研究发现,光动力过程中光剂量与光敏剂浓度存在一个最佳治疗剂量,其大小和组织中氧浓度有关,组织中越缺氧,最佳光动力剂量越小,光动力过程对组织的损伤越小,通过实验发现k562细胞悬浮液中最佳剂量为20×0.25(J/cm2·mmol/L),此时光动力损伤效果最明显.研究结果表明了PDT对肿瘤组织具有选择性光动力损伤的特点,并为PDT在临床上的广泛应用提供了理论支持.     

两种卟啉类光敏剂对SMMC-7721细胞敏化损伤的拉曼光谱分析

用拉曼光谱技术比较血卟啉单甲醚HMEM和血卟啉衍生物HpD对肝癌细胞SMMC-7721光敏化损伤。实验测定了用两种光敏剂对SMMC-7721细胞光动力作用后的DNA和蛋白质的光谱图。结果显示:HMME和HpD对DNA和蛋白质构象的光敏损伤特征是相同的,但损伤程度不同。经HMME作用的SMMC-7721细胞的谱线,DNA各峰的位置和强度的变化都比HpD大,有的峰消失,蛋白质主链的几条谱线强度与HpD谱线相比大幅降低,表示HMME对DNA和蛋白质主链的损伤较大。结果表明:HMME对SMMC-7721细胞的光敏     

竹红菌素修饰纳米脂质微泡联合LED光源照射对人舌癌细胞Tca8113的作用的实验研究

目的:本实验自制竹红菌素修饰的脂质微泡,联合LED光源对人舌癌细胞Tca8113进行光动力处理后观察细胞.方法:常规培养,取对数生长期的舌癌细胞Tca8113,机械振荡法制备竹红菌素修饰的脂质微泡,联合LED光源对细胞进行光动力治疗,MTT法检测细胞存活率,并用流式分析仪分析其促凋亡作用.结果:MTT法和流式细胞仪显示竹红菌素修饰的脂质微泡联合LED光源照射对Tca8113人舌癌细胞有明显凋亡现象,且在一定浓度范围和光照能量密度范围内,其凋亡作用与HB微泡浓度和能量密度呈正相关.结论:竹红菌素修饰的纳米脂质微泡联合LED光源光动力疗法对人舌癌细胞Tca8113有促凋亡抑制作用.     

光动力疗法中的治疗剂量学研究

为了研究光动力过程中光剂量和光敏剂剂量对光动力损伤效果的影响,基于肿瘤组织中光动力损伤剂量的数学模型以及光动力损伤剂量与组织中光剂量、光敏剂剂量、氧浓度之间的函数关系,用数学方法模拟研究了给定模型组织中光及光敏剂的有效吸收剂量与氧浓度之间的关系,并用ALA-PDT实验研究了K562细胞悬浮液中不同药物剂量对光动力损伤效...     

光敏剂亚细胞定位与诱导细胞死亡的信号途径

光动力疗法 (PDT)是一种能诱导细胞坏死和凋亡的细胞毒性治疗方法 ,光动力反应过程与许多信号途径有关 ,而光敏剂亚细胞定位是影响光动力杀伤效应的重要因素。光敏剂亚细胞定位后 ,在激发光作用下 ,通过各种信号途径诱导细胞坏死和凋亡。如 :光动力 -线粒体损伤 -细胞色素C释放 -caspase (白细胞介素 1 (转换酶 ) 3激活途径、光动力%D溶酶体原发损伤%D酸性鞘磷脂酶 (aSMase)活化%D神经酰胺 (Ceramide)释放 -细胞凋亡途径、光动力作用%D膜稳态失衡%D钙超载%D细胞死亡途径。目前 ,PDT引发的信号途径还未完全阐明 ,以往许多实验是以不同的模型和不同光敏剂为基础的 ,并且大多数实验是在体外进行的 ,许多共性的观点有待进一步确定 ,而一些信号途径实验依据较充分 ,在光动力诱导细胞死亡过程中发挥重要作用 ,实际上 ,这些途径常常综合起作用 ,体现了光动力作用机制的复杂性     

MPPA光动力作用诱导人鼻咽癌细胞凋亡的实验研究

为观察MPPa光动力作用对鼻咽癌细胞凋亡的影响,应用AnnexinV—PI双染结合流式细胞仪分析MPPa光动力作用后人鼻咽癌细胞株CNE2细胞发生凋亡和继发性坏死的比率。结果显示MPPa光动力作用实验组人鼻咽癌细胞株CNE2细胞发生凋亡和继发性坏死的比率分别增加到16.43 %和4.64 % ,且均显著高于单纯光照射组、单纯MMPa光敏剂处理组和假照射组(P <0 .0 1) ,而三对照组间无明显差异(P >0 .0 5 )。表明MPPa光动力作用能有效诱导低分化人鼻咽癌细胞株CNE2细胞凋亡的发生。这也可能是MPPa光动力作用杀伤鼻咽癌的重要机制之一。     

拉曼光谱研究不同剂量的光动力作用对肝癌细胞HepG2的杀伤效应

用拉曼光谱技术分析不同剂量的光动力作用对肝癌细胞HepG2的损伤.实验测定了4个光敏剂浓度组(光敏剂浓度为0.2、0.5、1.5、2.5μg/ml)和4个光剂量组(光剂量为6、12、18、24 J/cm2)及对照组HepG2细胞的拉曼光谱.结果表明,光剂量为6 J/cm2、光敏剂浓度为0.2μg /ml的谱线强度和峰位变化都较小,对HepG2细胞损伤较轻.随着剂量的增加各特征峰的强度变化率加大,细胞损伤加重,但光剂量高于18J/cm2、光敏剂浓度高于1.5μg /ml后,峰强变化率趋于平缓,到达平台期.提示光动力作用剂量存在有效杀伤阈值和安全阈值.     

A Self‐Transformable pH‐Driven Membrane‐Anchoring Photosensitizer for Effective Photodynamic Therapy to Inhibit Tumor Growth and Metastasis

Poor tumor selectivity and short life span of reactive oxygen species (ROS) are two major challenges in photodynamic therapy (PDT). In this study, a self‐transformable pH‐driven membrane anchoring photosensitizer (pHMAPS) is used to realize tumor‐specific accumulation and in situ PDT on tumor cell membrane to maximize the therapeutic potency. It is found that pHMAPS was able to form α‐helix structure under acidic condition (pH 6.5 or 5.5), while remain random coil at normal pH of 7.4. This pH‐driven secondary structure switch enables the successful insertion of pHMAPS into membrane lipid bilayer, especially for cancerous cell membrane in the acidic tumor microenvironment. Under laser irradiation, cytotoxic ROS is generated in the immediate vicinity of cell membrane, resulting in superior cell killing effect in vitro and significant inhibition of tumor growth in vivo. Importantly, benefited from this membrane‐specific PDT, tumor growth‐induced hepatic, pulmonary, as well as osseous metastases of breast cancer cells are also retarded after PDT treatment. Thus, the membrane localized PDT by pHMAPS provides a simple but effective strategy to enhance the medical performance of photosensitizing agents in cancer therapy.     

An O2 Self‐Sufficient Biomimetic Nanoplatform for Highly Specific and Efficient Photodynamic Therapy

Conventional oxygen‐dependent photodynamic therapy (PDT) has faced severe challenges because of the non‐specificity of most available photosensitizers (PSs) and the hypoxic nature of tumor tissues. Here, an O₂ self‐sufficient cell‐like biomimetic nanoplatform (CAT‐PS‐ZIF@Mem) consisting of the cancer cell membrane (Mem) and a cytoskeleton‐like porous zeolitic imidazolate framework (ZIF‐8) with the embedded catalase (CAT) protein molecules and Al(III) phthalocyanine chloride tetrasulfonic acid (AlPcS₄, defined as PS) is developed. Because of the immunological response and homologous targeting abilities of the cancer cell membrane, CAT‐PS‐ZIF@Mem is selectively accumulated at the tumor site and taken up effectively by tumor cells after intravenous injection. After the intracellular H₂O₂ penetration into the framework, it is catalyzed by CAT to produce O₂ at the hypoxic tumor site, facilitating the generation of toxic ¹O₂ for highly effective PDT in vivo under near‐infrared irradiation. By integrating the immune escape, cell homologous recognition, and O₂ self‐sufficiency, this cell‐like biomimetic nanoplatform demonstrates highly specific and efficient PDT against hypoxic tumor cells with much reduced side‐effect on normal tissues.     

曲古菌素A体外作用HeLa细胞的拉曼光谱分析

拉曼光谱是一种简便灵敏的光谱分析新技术,近年来在细胞的结构、功能等方面的研究取得了迅速发展.本文系统的分析了药物曲古菌素A作用于官颈癌细胞(HeLa细胞)24小时后的拉曼光谱变化,探讨了曲古菌素A作用后细胞内生物分子的结构及其含量的变化规律.结果表明:曲古菌素A作用后HeLa细胞中蛋白质的主链和侧链、核酸、脂类的谱带都...     

光动力治疗对荷瘤小鼠肿瘤淋巴细胞表型的影响

目的:观察光动力治疗(PDT)肿瘤留置对残存肿瘤局部淋巴细胞表型的影响。方法:采用双侧小鼠荷H22移植型肝癌模型。将30只双侧荷瘤小鼠分为3组:PDT组,切除组,对照组。光动力治疗一侧肿瘤后,定量观察残存肿瘤和脾脏内CD4+和CD8+细胞的变化。使用免疫组织化学及图像分析技术。结果:在原位肿瘤内,有极少量的CD4+细胞。在残存肿瘤内,未见CD4+细胞,CD8+细胞呈明显的带状分布,在光动力治疗组,CD8+细胞的密度明显增高,但仍不足以杀死残存肿瘤,其活性也有待进一步研究。     

光动力法制备抗小鼠H22肝癌的肿瘤疫苗

研究了光动力疗法(PDT)制备的抗小鼠H22肝癌肿瘤疫苗的抗瘤效应.将昆明鼠60只,随机分为2组,每组30只.实验组取6~12周龄的昆明鼠背部皮下接种光动力疗法产生的疫苗,每3天注射一次,每次注射50μL(相当于3×105个细胞),连续两周.隔一周于第22天注射H22肿瘤细胞悬液0.1 mL(1×106个细胞);对照组:每周每次注射50μL生理盐水,连续两周.隔一周于第22天注射H22肿瘤细胞悬液0.1 mL(1×106个细胞).比较两组的抑瘤率、生存率以及两组之间免疫学的相关指标.结果表明,实验组小鼠具有显著的抑瘤效果,抑瘤率、生存率较对照组有显著提高.实验组肿瘤抑瘤率最高可达60%且长期有效,100天生存率达56%.说明光动力疗法产生的抗小鼠H22肝癌疫苗可以有效地抑制肿瘤生长,提高荷瘤小鼠的生存率,具有明显的抗瘤效应.该方法可能成为一种辅助性治疗肿瘤的手段而应用于临床.     

Two-Photon Absorption Induced Cancer Immunotherapy Using Covalent Organic Frameworks

Immune checkpoint blockade therapy is revolutionizing the traditional treatment model of multiple tumor types, but remains ineffective for a large subset of patients. Photodynamic therapy (PDT) has been shown to induce cancer cell death and provoke an immune response, and may represent a potential strategy to synergize with immune checkpoint blockade therapy. However, the limited tissue penetration of exciting light for conventional PDT largely hinders its application in the clinic and its further combination with immunotherapy. Here, a serrated packing covalent organic framework (COF), COF-606, with excellent two-photon absorption (2PA) property and photostability, largely avoids aggregation-caused quenching, therefore offering high reactive oxygen species (ROS) generation efficiency; it is used as a 2PA photosensitizer for PDT in deep tumor tissue. COF-606 induced PDT is shown to be efficient in inducing immunogenic cell death, provoking an immune response and normalizing the immunosuppressive status for the first time. This makes it possible to combine 2PA induced PDT using COF with programmed cell death protein 1 immune checkpoint blockade therapy. Such combination leads to strong abscopal tumor-inhibiting efficiency and long-lasting immune memory effects, standing as a promising combinatorial therapeutic strategy for cancer treatment.     

A Near-Infrared Light-Responsive ROS Cascade Nanoplatform for Synergistic Therapy Potentiating Antitumor Immune Responses

Tumor occurrence is closely related to the unlimited proliferation and the evasion of the immune surveillance. However, it remains a challenge to kill tumor cells and simultaneously activate antitumor immunity upon spatially localized external stimuli. Herein, a robust tumor synergistic therapeutic nanoplatform is designed in combination with dual photosensitizers-loaded upconversion nanoparticles (UCNPs) and ferric-tannic acid (FeTA) nanocomplex. Dual photosensitizers-loaded UCNPs can induce photodynamic therapy (PDT) effect by generation of cytotoxic reactive oxygen species (ROS) on demand under near-infrared (NIR) light irradiation. FeTA can robustly respond to acidic tumor microenvironment to produce Fe²⁺ and subsequently induce chemodynamic therapy (CDT) by reacting with H₂O₂ in the tumor microenvironment. More importantly, the CDT/PDT synergy can not only exhibit significant antitumor ability but also induce ROS cascade to evoke immunogenic cell death. It increases tumor immunogenicity and promotes immune cell infiltration at tumor sites allowing further introduction of systemic immunotherapy responsiveness to inhibit the primary and distant tumor growth. This study provides a potential tumor microenvironment-responsive nanoplatform for imaging-guided diagnosis and combined CDT/PDT with improved immunotherapy responses and an external NIR-light control of photoactivation.     

FAP-Targeted Photodynamic Therapy Mediated by Ferritin Nanoparticles Elicits an Immune Response against Cancer Cells and Cancer Associated Fibroblasts

Cancer-associated fibroblasts (CAFs) are present in many types of tumors and play a pivotal role in tumor progression and immunosuppression. Fibroblast-activation protein (FAP), which is overexpressed on CAFs, has been indicated as a universal tumor target. However, FAP expression is not restricted to tumors, and systemic treatment against FAP often causes severe side effects. To solve this problem, a photodynamic therapy (PDT) approach is developed based on ZnF₁₆Pc-loaded and FAP-specific single chain variable fragment (scFv)-conjugated apoferritin nanoparticles, or αFAP-Z@FRT. αFAP-Z@FRT PDT efficiently eradicates CAFs in tumors without inducing systemic toxicity. When tested in murine 4T1 models, the treatment elicits anti-cancer immunity, causing suppression of both primary and distant tumors, that is, the abscopal effect. Treatment efficacy is enhanced when αFAP-Z@FRT PDT is used in combination with anti-PD1 antibodies. Interestingly, it is found that the PDT treatment not only elicits a cellular immunity against cancer cells, but also stimulates an anti-CAFs immunity. This is supported by an adoptive cell transfer study, where T cells taken from 4T1-tumor-bearing animals treated with αFAP PDT retard the growth of A549 tumors established on nude mice. Overall, this approach is unique for permitting site-specific eradication of CAFs and inducing a broad spectrum anti-cancer immunity.     

Red Blood Cell‐Facilitated Photodynamic Therapy for Cancer Treatment

Photodynamic therapy (PDT) is a promising treatment modality for cancer management. So far, most PDT studies have focused on delivery of photo­sensitizers to tumors. O₂, another essential component of PDT, is not artificially delivered but taken from the biological milieu. However, cancer cells demand a large amount of O₂ to sustain their growth and that often leads to low O₂ levels in tumors. The PDT process may further potentiate the oxygen deficiency, and in turn, adversely affect the PDT efficiency. In the present study, a new technology called red blood cell (RBC)‐facilitated PDT, or RBC‐PDT, is introduced that can potentially solve the issue. As the name tells, RBC‐PDT harnesses erythrocytes, an O₂ transporter, as a carrier for photosensitizers. Because photosensitizers are adjacent to a carry‐on O₂ source, RBC‐PDT can efficiently produce ¹O₂ even under low oxygen conditions. The treatment also benefits from the long circulation of RBCs, which ensures a high intraluminal concentration of photosensitizers during PDT and hence maximizes damage to tumor blood vessels. When tested in U87MG subcutaneous tumor models, RBC‐PDT shows impressive tumor suppression (76.7%) that is attributable to the codelivery of O₂ and photosensitizers. Overall, RBC‐PDT is expected to find wide applications in modern oncology.