光动力治疗裸小鼠胰腺移植癌的实验研究

目的 研究光动力治疗（ＰＤＴ）对胰腺癌的治疗效果,为胰腺癌寻找有效的治疗手段。方法 接种人胰腺癌细胞ＳＷ１９９０于裸小鼠皮下,建立移植癌模型,以血卟淋衍生物（ＨｐＤ）作为光敏剂,采用腹腔和瘤内局部注射两种不同方式给药,继以波长６３２．８ｎｍ的Ｈｅ－Ｎｅ激光照射肿瘤局部,观察光动力治疗后肿瘤的生长速度和组织形态学变化,并测定了肿瘤组织内脂质过氧化产物丙二醛（ＭＤＡ）的含量,对其治疗机制进行了初步探讨     

血卟啉光动力作用杀伤人胆管癌细胞的实验研究

目的研究血卟啉化衍生物（hematoporphyrin derivative，HPD）光动力作用（photodynamic therapy，PDT）对人胆管癌细胞QBC939的杀伤效应：方法以人胆管癌细胞系QBC939为研究对象，采用血卟啉化合物作为光敏剂，半导体激光治疗仪为光源。实验分为对照组（空白对照组、单纯光动力组、单纯光敏剂组）和光动力作用组，以系列浓度的血卟啉经不同强度光照后，用MTT法测定PDT对QBC939细胞的相对抑制率和筛选最佳PDT参数。结果光动力作用组的吸光度（OD值）与对照组间的差异具有统计学意义（P〈0.01），随着HPD浓度的增加和光照强度的增大，光动力作用对细胞的相对抑制率逐渐增大。不同的光敏剂浓度和光照强度间组合能达到相同的抑制率，如以10mg／L HPD经5J／cm^2光照强度或以4mg／L HPD经15J／cm^2光照强度对QBC939均可以达到92％相对抑制率。在最佳光照能量密度条件下，设计三组功率-时间组合，组间没有显著差异。结论1）PDT对人胆管癌细胞QBC939具有明确的杀伤作用，其对细胞的抑制率具有显著的剂量效应关系。光敏剂浓度和光照强度间存在交互关系，从临床角度考虑，采用较低的光敏剂浓度经较大的光照强度照射是理想的PDT治疗方案。2）改变功率时间的组合不会影响光动力对胆管癌细胞杀伤作用，采用在光纤承受范围内的大功率短时间的照射方式可达到安全快捷的目的。     

光动力治疗诱导人胰腺癌细胞凋亡的实验研究

目的 研究光敏剂血卟啉光动力作用对人胰腺癌细胞Panc-1的体外杀伤效应及其主要机制。方法 将光敏剂浓度、光照剂量两个因素按不同水平分组,以CCK-8实验的OD值为检测指标并转换为细胞存活率,研究两个因素对光动力作用的影响及其规律。在此基础上,依次以透射电镜、荧光显微镜、流式细胞仪测定不同处理强度的光动力作用后细胞凋亡及坏死的特点,探讨光动力杀伤肿瘤细胞的主要机制。结果 随着光敏剂浓度和光照剂量的增加,PDT后Panc-1细胞存活率相应下降,但单独给予光敏剂和光照均不对细胞存活率产生影响。PDT后细胞出现凋亡和坏死,二者比例随光敏剂浓度和光照剂量的增加而增加,但始终表现为凋亡率＞坏死率。结论 血卟啉光动力治疗对人胰腺癌细胞株Panc-1具有明确的杀伤作用,但是光敏剂和激光照射本身并不具有独立的杀伤效应。光敏剂浓度、光照剂量两个影响因素在一定范围内与PDT效应之间成正相关的关系。PDT破坏肿瘤细胞的作用机制主要在于诱导细胞凋亡。     

光动力学疗法联合局部化疗治疗上消化道肿瘤的护理

光动力学疗法（ＰＤＴ）是利用光敏剂够够选择性地聚积滞留于肿瘤组织，产生光动力杀伤效应，达到局部抗肿瘤作用［１］。本科１９９５～１９９８年对４１例进展期食管贲门癌病人进行治疗，护理体会如下。１　临床资料４１例病人中，男３３例，女８例，年龄４４～８１岁，平均５８．６岁。食管鳞状细胞癌２７例，其中手术及放疗后复发各２例；贲门癌１４例，术后复发５例，其中腺癌１例，鳞癌２例，印戒细胞癌１例，腺鳞癌１例。２　材料与方法２．１　材料：光敏剂为ＰＳＤ００７注射液，每安瓿１００ｍｇ／１０ｍｌ，上海第二军医大学５…     

胰腺癌细胞对化疗药物敏感性的测定

目的 探讨人胰腺癌细胞株对常用化疗药物及其相互配伍的敏感性，为临床合理用药提供理论依据。方法 将人胰腺癌细胞株（Ｐ３、ＳＷ１９９０、Ｃａｐａｎ－２）细胞接种于９６孔培养板，分别加入不同配伍及不同浓度的化疗药物，作用一定时间后采用噻唑蓝法（ＭＴＴ法）测定其抗癌敏感性。结果 以５－氟脲嘧啶和丝裂霉素为基本的三联用药在抑制率相当的情况下，其组成中各药物剂量仅为单一用药剂量的１／５０；联合用药方案中以５－     

2-丁胺-2-去甲氧基竹红菌乙素光动力作用诱导Capan-1细胞凋亡与线粒体损伤的关系

目的 研究新型光敏剂2-丁胺-2-去甲氧基竹红菌乙素（BAHB）的光动力作用诱导人胰腺癌细胞系（Capan-1）凋亡的机制。方法 利用荧光显微镜、细胞凋亡DNA梯带、激光共聚焦扫描显微镜、Westernblot分析等技术研究BAHB诱导Capan-1凋亡与线粒体损伤的关系。结果低浓度BAHB主要定位于线粒体,高浓度时定位于溶酶体。DNA琼脂糖电泳分析显示出特征性细胞凋亡的DNA梯带。激光共聚焦扫描显微镜观察到线粒体跨膜电位（△ψm）降低,定量分析结果表明,随着光照时间的缩短,细胞的相对荧光强度增强。用Westernblot法可以检测到线粒体中细胞色素C的释放。结论 低浓度BAHB的光动力作用诱导人胰腺癌细胞凋亡的机制与线粒体损伤有关。     

膀胱癌化疗药物敏感性的实验研究

以膀胱癌细胞株（ＢＩＵ－８７）为靶细胞，采用噻唑蓝（ＭＴＴ）比色法进行了膀胱癌腔内化疗药物敏感试验及条件选择的实验研究。结果发现丝裂霉素Ｃ（ＭＭＣ）、噻替哌（ＴＨＴ）、顺铂（ＤＤＰ）对ＢＩＵ－８７细胞杀伤作用较明显，但在１ｍｇ／ｍｌ、２ｈ作用条件下均未达到５０％抑制率、需增加药物浓度和延长作用时间。认为ＭＭＴ、ＴＨＴ、ＤＤＰ对ＢＩＵ－８７细胞的毒性作用与药物浓度和作用时间有关；为达到最大疗效，药物     

肝癌病人树突状细胞诱导高效而特异的抗肝癌免疫

目的 以肝癌病人树突状细胞（ＤＣ）体外诱导抗肝癌免疫。方法 自肝癌病人外周血中分离出单个核细胞（ＰＢＭＣ０;以人肝癌细胞系ＨｅｐＧ２肿瘤细胞的肿瘤相关抗原（ＴＡＡ）激活ＤＣ;以粒／巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）及白介素４（ＩＬ－４）联合刺激ＰＢＭＣ中ＤＣ;ＤＣ诱导自体Ｔ淋巴细胞增殖,分化为细胞毒性Ｔ细胞（ＣＴＬ）;检测ＣＴＬ及其上清液对ＨｅｐＧ２,ＢＥＬ－７４０２,ＬＯＶＯｅｙ ＨＯＳ－８６     

光动力治疗裸鼠胰腺癌及其机理的实验研究

目的 探讨在裸鼠胰腺癌瘤内注射光敏剂血卟啉衍生物（HpD）、竹红菌甲素（HA）及2－丁胺－2－去甲氧基竹红菌甲素（2－BA－2－DMHA）后，光动力治疗（PDT）的效果及其作用机理。方法 将人胰腺癌细胞株SW1990接种到裸鼠皮下，建立胰腺癌动物模型，而后将光敏剂HpD、HA及2－BA－2－DMHA注射入肿瘤，再用激光照射肿瘤局部，观察PDT治疗胰腺癌的效果。另以第Ⅷ因子为标记，采用免疫组化LSAB法研究肿瘤血管损伤的情况。结果 PDT对胰腺肿瘤有明显杀伤作用，能使胰腺肿瘤生长速度明显减慢。免疫组化染色明显，PDT可引起血管内皮细胞损伤和血管结构破坏。结论 PDT对胰腺癌有治疗作用，血管损伤是其杀伤肿瘤的重要途径。     

脂质体介导c-myc基因反义寡核苷酸对人肝癌细胞的生物学影响

目的 探讨应用脂质体（ＬＲ）介导ｃ－ｍｙｃ基因反义寡核苷酸（ＡＳＯＤＮ）对人肝癌细胞ＳＭＭＣ－７７２１（７７２１细胞）的生物学影响。方法 利用１ｍｇ／Ｌ ＬＲ包裹２μｍｏｌ／Ｌ ｃ－ｍｙｃ ＡＳＯＤＮ转染于培养的人肝癌细胞,观察其对瘤细胞的作用。结果 四唑盐比色实验（ＭＴＴ）法测定ＬＲ－ＡＳＯＤＮ组作用４８ｈ对瘤细胞生长抑制率（５５％）较无ＬＲ的ＡＳＯＤＮ组的生长抑制率（１５％）显著提高（Ｐ〈０．     

金属卟啉化合物对人前列腺癌细胞PC-3的光动力杀伤作用

目的:研究水溶性金属卟啉化合物对人前列腺癌细胞PC 3的光动力学杀伤作用,探讨金属卟啉的光动力 学杀伤作用抗肿瘤的机制。　方法:合成水溶性金属卟啉化合物5,10,15,20 四(N 甲基 4 吡啶基)锰卟啉四碘盐 后,采用四甲基偶氮唑蓝比色法和膜联蛋白/碘化丙锭双标记流式细胞术分别检测不同浓度的金属锰卟啉化合物 (0、0.1、1、10μmol/L)在不同时间(0、15、30、45min)的高压汞灯(40W)照射下,对PC 3细胞的生长活性和凋亡的 影响。　结果:锰卟啉浓度在0～10μmol/L、光照0～30min范围内,其对PC 3细胞的增值抑制作用和诱导PC 3 细胞的凋亡作用均随着浓度增加和光照时间的增加而增强,但大剂量(10μmol/L)、高能量照射(光照45min)时主 要导致PC 3细胞坏死。　结论:水溶性金属卟啉化合物5,10,15,20 四(N 甲基 4 吡啶基)锰卟啉四碘盐的光动 力作用在体外能诱导PC 3细胞凋亡,有效杀伤PC 3细胞,其杀伤效果在锰卟啉浓度为0～10μmol/L、光照0～30 min范围内与锰卟啉的浓度和光照射剂量呈正相关,这可能是卟啉的光动力学疗法抗肿瘤作用的机制。     

Selective necrosis in hamster pancreatic tumours using photodynamic therapy with phthalocyanine photosensitization.

Photodynamic therapy (PDT) is often thought to be able to effect selective tumour necrosis. This therapeutic selectivity, based on transient differences in tumour: normal tissue photosensitizer concentration ratios, is rarely useful clinically in extracranial tumours, although PDT itself may be of value by virtue of the nature of the damage produced and healing of normal tissue by regeneration. This report describes the effects of PDT on normal pancreas and chemically induced pancreatic cancers in the hamster, where a different mechanism of selective necrosis may be seen. Photosensitizer distribution in normal and neoplastic pancreas was studied by chemical extraction and fluorescence microscopy. Correlation of distribution studies with necrosis produced by PDT shows that the photodynamic dose (product of tissue concentration of sensitizer and light dose) threshold for damage is seven times as high for normal pancreas as for pancreatic cancer. Tumour necrosis extended to the point where tumour was invading normal areas without damaging the normal tissue. In rat colonic cancer, photodynamic dose thresholds in tumour and normal tissue are similar and so such marked selectivity of necrosis is not possible. The reason for this selectivity in the pancreas is not clear, but recent evidence has suggested a difference in response to PDT between normal and neoplastic pancreatic cell lines and the presence of a singlet oxygen scavenger in normal pancreas is postulated. Furthermore, the present fluorescence microscopy studies suggest that tumour stroma contains the highest level of photosensitizer and thus receives the highest photodynamic dose during PDT. These results suggest a possible role for PDT in treating small pancreatic tumours or as an adjuvant to other techniques, such as surgery, that reduce the main bulk of tumours localized to the pancreas.     

An ultrastructural study of human bladder cancer treated by photodynamic therapy

An electron microscopic study has been carried out biopsy material of transitional cell carcinoma of human bladder, taken before and after photodynamic therapy (PDT), from 14 patients. It has been demonstrated that bladder cancer is highly sensitive to PDT using haematoporphyrin derivative (HPD, made in China) and laser irradiation. It was found that vascular endothelium within the tumour tissue was very sensitive to PDT, showing distinct changes as early as immediately after completion of a 20-min irradiation. Twenty-four hours after PDT, almost all the capillaries examined were necrotic and broken down into small fragments. The cancer cells were less sensitive to PDT, being damaged later and less seriously than the blood vessels. It has been concluded that in PDT-treated tumours the vascular endothelium is damaged primarily while the cancer cells are destroyed, to a considerable extent, secondarily as the consequence of structural damage to capillaries and functional disturbance in the microcirculation. We would speculate that the main factor influencing the final response is the actual concentration of HPD in various types of cell in the tumours.     

Photodynamic therapy for head and neck cancer xenografts in athymic mice

This study examines efficacy and optimal treatment variables of photodynamic therapy (PDT) for human head and neck squamous cancer (HNSC) xenografts in athymic mice. Two and four days after injection of hematoporphyrin derivative (HPD), tumors were illuminated with red light from an argon-dye laser. Sixty-three tumors were treated. With HPD dose and light intensity constant at 7.5 mg/kg and 100 mW/cm2, respectively, the extent of tumor necrosis was strongly dependent on duration of light exposure. There was no substantial difference in results for 30- and 60-minute treatment durations between animals injected with HPD 2 and 4 days before treatment. After 30 minutes treatment time, responses were seen in 8 of 10 mice (2 days post-HPD) and 11 of 12 mice (4 days post-HPD). After 60 minutes treatment time, toxicity was high. We conclude that, in this model, PDT is effective in selective killing of HNSC. For future comparison studies in this model, if the indicated HPD dose and light intensity are used we recommend a 2-day delay after HPD injection and a light exposure duration of 30 minutes.     

5-氨基乙酰丙酸光动力学疗法对两种人卵巢癌细胞的体外杀伤作用

目的：探讨5-氨基乙酰丙酸（5-ALA）光动力学疗法（photodynamic therapy,PDT）对体外培养人卵巢癌SKOV3和AO细胞的杀伤作用。方法：于体外培养的SKOV3和AO细胞分别加入不同浓度的5-ALA（0．1、0．5、1．0、2．5、12．5mmol／L）孵育4h,以波长为630nm的半导体激光进行照射,照射能量密度分别为0．1、0．5、2．5、12．5J／cm^2。用四唑盐（MTT）比色法测定细胞存活率,并与单纯药物对照组（不同浓度5ALA孵育、无光照）、单纯光照组（无5-ALA孵育、不同能量密度光照）与阴性对照组（无药物孵育、无光照）进行比较。计算5ALA—PDT对SKOV3和AO细胞的半数杀伤浓度（IC50）。结果：5-ALA—PDT处理后两种卵巢癌细胞存活率均明显下降（P〈0．05）,提示5ALA—PDT对体外培养的人卵巢癌SKOV3和AO细胞均有杀伤作用,其杀伤作用随5-ALA的浓度和激光能量密度增加而增加,当5-ALA浓度超过2．5mmol／L或照射能量密度超过2．5J／cm^2时细胞存活率变化不明显。单纯药物对照组、单纯光照组的细胞存活率与阴性对照组相比差异无显著性（P〉0．05）。5-ALA-PDT对两种卵巢癌细胞杀伤效应强弱不同,对SKOV3及AO细胞的半抑制浓度（IC50）分别为0．34mmol／L和2．50mmol／L,两者差异有显著性（P〈0．05）。结论：5-ALA-PDT可有效杀伤体外培养的人卵巢癌细胞SKOV3和AO细胞,且对两种细胞的杀伤作用有差异,对SKOV3细胞株的杀伤效应要强于AO细胞株。     

Photodynamic therapy: Optimal treatment parameters in murine fibrosarcoma

We have investigated the efficacy of photodynamic therapy (PDT) by using two preparations of haematoporphyrin derivative (HPD) that demonstrated different biological activities against experimental murine fibrosarcoma RIF-1 in C3H/He mice. We have been able to demonstrate a minimum clonogenic survival rate of 0.25% by using the more active HPD at 20 mg/kg with a 40-h retention time and a total laser light dose of 100 J/cm². Further, we noted that clonogenic survival rates of 21.6% and 25.0% respectively could be achieved by using the less active HPD (at 20 mg/kg) with a laser light dose of 150 J/cm², or the more active HPD (at 20 mg/kg) with a laser light dose of 20 J/cm². In both cases necrosis of the surrounding normal tissue was absent. Necrosis of the normal tissue surrounding the tumour was shown to be associated with high laser energy (100 J/cm² and higher) in conjunction with a high dose (20 mg/kg) of the more active HPD. A comparison of survival curves as a function of laser energy for the RIF-1 cells following PDT with the two different preparations of HPD showed a difference in the kinetics of cell death. A curve with a shoulder region and aD _o (mean lethal dose) of 41 J/cm² was obtained when the less active HPD was used, whereas PDT using the more active HPD resulted in a curve with no shoulder and aD _o of 16 J/cm².     

Human pancreatic carcinoma cells are sensitive to photodynamic therapy in vitro and in vivo.

BACKGROUND: The aim of this study was to assess the efficiency of photodynamic therapy (PDT) on human pancreatic cancer cells in vitro and in an animal model. METHODS: Human pancreatic tumour cell lines were submitted to PDT with pheophorbide a (Ph a), a chlorophyll derivative, in culture and after grafting into athymic mice. Ph a was tested in culture (10-10-10-5 mol/l) with a 5-J/cm2 energy treatment and on tumour-bearing Nude mice (30 mg/kg intraperitoneally) with a 100-J/cm2 PDT session. The effect of PDT was assessed in vitro using proliferative, apoptotic and clonogenic tests and in vivo on tumour growth and on the induction of tumour necrosis. RESULTS: PDT inhibited tumour cell growth in culture by affecting DNA integrity. This tumour cell photodamage started at low concentration (10-7 mol/l) as corroborated by clonogenic and tumour growth tests. A strong necrosis was achieved in vivo with a single PDT session. CONCLUSION: PDT destroyed human pancreatic carcinoma after low photosensitizer supply and weak energy application. It exerted this tumoricidal effect via apoptosis induction with a gentle protocol, and apoptosis and/or necrosis with a stronger protocol.     

Hyperthermia potentiates the effects of aluminum phthalocyanine tetrasulfonate-mediated photodynamic toxicity in human malignant and normal cell lines.

The purpose of this study was to examine the effects of photodynamic therapy utilizing aluminum phthalocyanine tetrasulfonate in vitro on several human malignant and normal cell types, with or without hyperthermia. Cells examined included normal skin fibroblasts, HT-1080 fibrosarcoma cells, SCC-25 (squamous cell carcinoma) and malignant melanoma cells. An argon-pumped continuous wave tunable dye laser at 675 nm was used as the light source, hyperthermia groups were heated to 42.5 degrees C, and radioisotope incorporation was used to measure DNA and protein synthesis as toxicity assays. Results showed an energy-dose, and A1PcS-concentration dependent toxicity in all cell lines examined, with moderate selectivity toward malignant cells. Hyperthermia alone was slightly toxic in melanomas and HT-1080 cell lines but had no effect in normal fibroblasts or SCC-25 cells. Hyperthermia synergistically potentiated the effects of PDT in all cell lines, and the combined modality was significantly more toxic in all malignant cell lines compared with normal cells. Thus, addition of hyperthermia to PDT protocols may enhance the efficacy of this treatment modality in vitro.