热疗联合榄香烯对SMMC 7721肝癌细胞HSP70的影响

目的探讨榄香烯联合热疗对肝癌细胞系SMMC 7721热休克蛋白70(HSP70)表达的变化以及与细胞凋亡的关系。方法采用肝癌细胞系SMMC 7721体外培养,MTT法测定单纯热疗、单纯榄香烯及榄香烯联合热疗对细胞增殖的影响。另外用流式细胞术的方法测定细胞凋亡与HSP70的表达。结果经热处理、榄香烯处理及联合处理,对肝癌细胞的生长均有明显的抑制作用,并诱导产生细胞凋亡。单纯热疗与单纯榄香烯处理细胞后HSP70表达增加,而榄香烯联合热疗后HSP70表达与前两组有所下降,细胞凋亡率却有所增加,与前两组比较差异有统计学意义(P<0.01)。结论榄香烯联合热疗下调了HSP70的表达,增加了细胞的凋亡率。     

鸦胆子油乳注射液联合热疗对人膀胱癌细胞株BIU-87细胞增殖及p-Akt表达的影响

目的探讨鸦胆子油乳注射液(简称鸦油乳)联合热疗对人膀胱癌细胞株BIU-87膀胱癌细胞形态、增殖、细胞周期及磷酸化丝氨酸苏氨酸蛋白激酶(p-Akt)蛋白表达的影响。方法采用倒置光学显微镜观察鸦油乳(10 ml/L)与热疗单独或联合对人BIU-87膀胱癌细胞形态学的影响;用四甲基偶氮唑蓝(MTT)法检测鸦油乳(5、10、20、30 ml/L)与热疗单独或联合对人BIU-87膀胱癌细胞增殖的抑制作用;采用流式细胞仪检测鸦油乳(10 ml/L)与热疗单独或联合对人BIU-87膀胱癌细胞周期的影响;用蛋白质印迹法(Western blotting)检测鸦油乳(10 ml/L)与热疗单独或联合作用于人BIU-87膀胱癌细胞24 h后p-Akt蛋白的表达。结果倒置光学显微镜结果显示,热疗组、鸦油乳组和鸦油乳联合热疗组的细胞数均明显减少,部分细胞变圆、体积变小、核固缩,但鸦油乳联合热疗组变化最明显,活细胞数最少。与热疗组和鸦油乳组比较,鸦油乳联合热疗对BIU-87膀胱癌细胞增殖的抑制作用显著增强(P<0.05),呈时间和剂量依赖性;鸦油乳联合热疗作用于人BIU-87膀胱癌细胞24、48、72 h的半数抑制浓度(IC50)分别为84、62、35 mg/L,低于单药鸦油乳的101、79、57 mg/L。鸦油乳联合热疗组作用24 h后人BIU-87膀胱癌细胞S期比率为(16.79±2.78)%,显著低于热疗组的(29.52±3.31)%和鸦油乳组的(26.34±0.86)%(P<0.05)。鸦油乳联合热疗组p-Akt蛋白的相对表达量为(0.51±0.02),显著低于热疗组的(0.98±0.05)和鸦油乳组的(0.70±0.04)(P<0.05)。结论鸦油乳联合热疗能增强对人BIU-87膀胱癌细胞的增殖抑制作用,下调p-Akt蛋白表达可能是其作用机制之一。     

水芹总酚酸对人肝癌2.2.15细胞周期的影响

目的研究水芹总酚酸对人肝癌2.2.15细胞增殖抑制及细胞周期的影响。方法用不同浓度的水芹总酚酸作用于体外培养的2.2.15细胞,采用MTT法检测水芹总酚酸对2.2.15细胞增殖抑制能力;倒置显微镜观察2.2.15细胞的形态学变化;流式细胞术检测水芹总酚酸对2.2.15细胞周期的影响。结果水芹总酚酸对2.2.15细胞增殖具有明显的抑制作用,且具有剂量依赖性;流式细胞术检测表明水芹总酚酸作用于2.2.15细胞后,可将细胞阻滞在S期。结论水芹总酚酸对人肝癌2.2.15细胞具有明显的抑制增殖作用,可能与阻滞细胞周期有关。     

葡萄籽原花青素联合顺铂对喉癌细胞Hep-2生物学作用及机制研究

目的利用葡萄籽原花青素及其联合顺铂在体外作用于喉癌细胞Hep-2,观察其对喉癌细胞增殖、凋亡的影响,并探讨抗肿瘤作用的可能机制。方法 MTT法检测各组药物对喉癌细胞增殖的抑制作用;流式细胞仪检测细胞周期;TUNEL法检测各组药物对喉癌细胞凋亡率的影响;采用流式细胞仪分析凋亡细胞比例,Western blot法检测各组药物对喉癌细胞Bax/Bcl-2表达情况的影响。结果 MTT和TUNEL结果显示,GSPE+顺铂组的喉癌细胞抑制增殖作用明显高于单纯GSPE组和单纯顺铂组,差异均有统计学意义(P<0.05);细胞周期结果显示,GSPE+顺铂组处于S+G2M期的细胞数明显低于单纯GSPE和单纯顺铂组,差异均有统计学意义(P<0.05);蛋白表达结果显示,GSPE+顺铂组的Bax/Bcl-2的表达明显高于单纯GSPE组和单纯顺铂组,差异均有统计学意义(P<0.05)。结论葡萄籽原花青素可诱导喉癌Hep-2细胞凋亡;与单纯应用葡萄籽原花青素或顺铂比较,葡萄籽原花青素联合顺铂可以起到更好的增殖抑制并诱导喉癌细胞凋亡,两者具有协同作用。     

模拟失重对胃粘膜上皮细胞增殖和周期的影响

目的 探讨模拟失重对胃粘膜上皮HFE-145细胞增殖和周期的影响.方法 采用回转器模拟失重,用免疫组化法观察HFE-145细胞PCNA表达的变化,用流式细胞仪测定细胞周期.结果 回转器模拟失重后,与正常对照组相比,回转组HFE-145细胞24、36、48及72 h的PCNA抗体阳性细胞比率无明显差异,12 h回转组阳性细胞比率明显减少（P〈0.05）.24、36、48、72 h回转组HFE-145细胞的细胞周期分布与对照组无明显差异,12 h回转组G0＋G1期细胞比例明显增多（P〈0.05）,S＋G2＋M期细胞之和显著减少（P〈0.05）.结论 回转器模拟失重使胃粘膜上皮HFE-145细胞在12 h时出现了细胞周期阻滞和增殖抑制.     

端粒酶反义寡核苷酸对恶性脑胶质瘤细胞生长的抑制作用

为观察端粒酶反义寡核苷酸(ODN)对恶性脑胶质瘤细胞的生长抑制作用，采用手术切除的端粒酶活性呈阳性奉达的Ⅲ、Ⅳ级脑胶质瘤标本16例，提取细胞进行原代培养后，以5μmol/L端粒酶反义寡核苷酸与恶性胶质瘤细胞进行孵育，每隔24h取样，以流式细胞仪检测PCNA、TUNEL阳性细胞百分率及细胞周期时相。结果显示，反义寡核苷酸对端粒酶活性于48h出现抑制，72h完全抑制，72h后对胶质瘤细胞增殖出现显著抑制并对细胞凋亡有显著促进作用，96h后细胞多滞留于G2／M期，此期凋亡细胞百分率随时间延长而增加。提示端粒酶反义寡核苷酸对端粒酶活性具有抑制作用，并可引起细胞周期阻滞，抑制脑胶质瘤细胞增殖而促进其凋亡。     

索拉非尼联合顺铂抑制肝癌HepG2细胞生长的体外研究

目的探讨索拉非尼联合顺铂(DDP)对肝癌细胞HepG2生长的抑制作用及其可能的分子机制。方法实验细胞分为4组:空白对照组、索拉非尼单药组、顺铂单药组、索拉非尼联合顺铂组。索拉非尼和顺铂单独或联合作用于细胞,以MTT法检测24、48、72、96h时间点药物对HepG2细胞的增殖抑制率,流式细胞仪检测24、48h时间点的细胞周期与凋亡率。在药物作用后24h,用荧光染料罗丹明123(Rh123)染色细胞,流式细胞仪测定线粒体跨膜电位(ΔΨm),分光光度法检测caspase-3活性。结果索拉非尼及顺铂单独应用对HepG2生长均有抑制作用,且低剂量联合具有协同作用。联合用药可以使细胞周期阻滞于G0/G1期和G2期,且凋亡率明显高于单药组(P<0.05)。单独应用顺铂和索拉非尼均能使线粒体跨膜电位降低,而联合组作用强于单药组(P<0.05)。药物作用后caspase-3活性增高,联合组高于单药组(P<0.05)。结论索拉非尼联合顺铂能更好地抑制肝癌HepG2细胞增殖,并促进其凋亡,其机制可能与细胞周期阻滞、线粒体跨膜电位降低及caspase-3活性增加有关。     

异鼠李素对人肝癌HepG-2细胞增殖与凋亡影响的实验研究

目的：观察异鼠李素对人肝癌HepG-2细胞的增殖周期及凋亡的影响。方法噻唑蓝（Mar）法检测异鼠李素对肿瘤细胞生长的抑制作用,流式细胞仪检测异鼠李素对肿瘤细胞周期及凋亡的影响。结果MTr结果提示异鼠李素对人肝癌细胞的增殖有明显抑制作用,并随着作用浓度的增大,抑制作用逐渐增强,以100mg／L的异鼠李素培养基抑制作用最明显。流式细胞仪检测可见亚二倍体峰,提示异鼠李素有诱导入肝癌细胞凋亡作用。异鼠李素可通过诱导人肝癌细胞凋亡并将细胞阻滞在G0—G1期,使细胞不能进入s期进行DNA合成,最终使瘤细胞的体外增殖受到抑制。结论异鼠李素抑制人肝癌细胞的增殖。阻滞细胞周期,诱导细胞凋亡,可能具有一定抗肝癌作用。     

全身放射损伤对皮肤伤口成纤维细胞增殖的影响

目的 研究合并全身放射损伤时创伤局部成纤维细胞数量减少与细胞增殖的关系。方法 采用组织学和免疫组化方法检测大鼠皮肤伤口局部成纤维细胞数量变化及与细胞增殖密切相关的增殖性细胞核抗原（PCNA）、细胞周期素E（Cyclin E)及细胞周期素依赖性激酶4（CDK4）含量变化。结果 合并全身放射损伤时皮肤伤口局部成纤维细胞数量在伤后5、7和10d显著少于单纯创伤组，并显著延迟高峰期；伤口局部PCNA和CDK4含量在务后3、5和7d显著低于单纯创伤组，伤口局部Cyclin E含量在伤后3、5、7和10d也显著低于单纯创伤组。结论 全身放射损伤可抑制伤后早期细胞周期G1期到S期的转变，抑制细胞增殖，从而使伤口局部成纤维细胞数量减少，这可能是合并全身放射损伤时创伤难愈的重要原因。     

热疗联合柔红霉素对CD34+CD38-白血病细胞增殖和凋亡的影响

目的 观察热疗联合柔红霉素(DNR)对CD34+ CD38-急性髓白血病KG1a细胞增殖的抑制作用和对其凋亡的影响.方法 免疫磁珠分离KG1a细胞中的CD34+ CD38- 细胞,以DNR作用48h后抑制50%CD34+ CD38- 细胞生长的药物浓度(IC50)作为后续实验的工作浓度.将CD34+CD38- 细胞分为对照组、热疗组(40℃、42℃)、化疗组和热化疗组(40℃、42℃).热疗、热化疗组在恒温水浴箱(40℃或42℃)hu加热60min,化疗组、热化疗组以IC50浓度的DNR处理48h.MTT法检测DNR对CD34+CD38- KG1a细胞增殖的抑制作用;流式细胞仪检测细胞凋亡率及42℃条件下细胞内DNR平均荧光强度的变化;甲基纤维素培养体系分析42℃条件下细胞的克隆形成能力.结果 KG1a细胞中分离出的CD34+CD38- 细胞纯度达95%以上,DNR的IC50为0.40μg/ml.热疗组、化疗组、热化疗组CD34+CD38-细胞增殖均明显受抑,且抑制作用随温度升高而增强(P<0.05).热疗组、化疗组及热化疗组细胞凋亡率均较对照组显著增加,且随温度升高凋亡率增加(P<0.05),热化疗组较化疗组及相同温度热疗组的细胞凋亡率显著增加(P<0.05).42℃热化疗组细胞内的DNR荧光强度(6.74±0.58)明显高于对照组(0.26±0.05)及化疗组(2.08±0.14,P<0.05).42℃热疗组、化疗组、42℃热化疗组细胞的集落形成率明显低于对照组,且42℃热化疗组明显低于42℃热疗组和化疗组(P<0.05).结论 热疗能增强DNR抑制CD34+ CD38- KG1a细胞增殖的作用,使细胞凋亡增多.     

Interaction between low dose-rate irradiation, mild hyperthermia and low-dose caffeine in a human lung cancer cell line.

PURPOSE: To investigate cell killing by means of low dose-rate irradiation (LDRI) combined with concurrent mild hyperthermia and to determine the effect of low-dose caffeine on this combination treatment. MATERIALS AND METHODS: Human lung adenocarcinoma cells, LK87, were treated with LDRI (50 cGy/h) in combination with mild hyperthermia at 41 degrees C and low-dose caffeine (1 mM). Cell survival was estimated by clonogenic assay. Flow-cytometry was performed with PI staining using FACScan. Heat-shock protein (HSP72/73) was measured by the Western blotting method. All treatments were simultaneously performed for up to 48 h (24 Gy). RESULTS: LDRI cytotoxicities were enhanced by hyperthermia at 41 degrees C. D0 calculated from the dose-response curve for LDRI combined with 41 degrees C was 3.46 Gy whereas it was 6.55 Gy for LDRI alone. The survival curve for LDRI +41 degrees C demonstrated no chronic thermotolerance up to 48 h. For LDRI + simultaneous low-dose caffeine, cell killing was also enhanced, where D0 was 3.38 Gy at 37 degrees C. Radiosensitization caused by caffeine was enhanced by combination with simultaneous mild hyperthermia at 41 degrees C, where D0=1.78 Gy. Cell cycle analysis demonstrated remarkable G2 and mild G1 arrest for LDRI alone, but only G1 arrest was observed for LDRI combined with 41 degrees C and for LDRI combined with caffeine. Strong and early G1 arrest was observed in the treatment with LDRI + caffeine at 41 degrees C. The amount of HSP72/73 in the combination of LDRI with caffeine at 41 degrees C was less than that at 41 degrees C alone. CONCLUSION: LDRI cytotoxicity was enhanced by non-lethal hyperthermia. Low dose caffeine produced further cell killing in the combination of LDRI with mild hyperthermia.     

Effect of adriamycin combined with 40 and 43 degrees C hyperthermia on cultured V79 cells and S-180 tumor and their uptake of the drug

The combined effects of hyperthermia and chemotherapy with adriamycin (ADR) on both cultured mammalian cells and growing tumors were analyzed according to different temperatures and durations of heating, timing of drug administration, and fractionation of treatments. Furthermore, ADR uptake in tumor was measured to evaluate the effect of ADR in combination with hyperthermia. Hyperthermic cell killing with ADR in vitro was much more potent at 40 degrees C than at 43 degrees C. Results of sequential combinations heating followed by ADR and ADR followed by heating under hyperthermia of 40 and 43 degrees C were not significantly different from those of simultaneous treatment. However, the anti-tumor effect of the combination was much more pronounced at 43 degrees C than at 40 degrees C for up to 60 min of treatment. ADR uptake showed a substantial increase under high temperatures of 40 and 43 degrees C in both cultured cells and tumors. Fractionated treatments combined with chemotherapy and hyperthermia showed no enhanced effects on tumor under temperatures of 40 and 43 degrees C, since three fractionated experiments with ADR and 43 degrees C hyperthermia had an effect almost identical to that of three fractionated experiments with hyperthermia alone, suggesting the possible induction of thermotolerance by ADR. The results of the present study indicate that potentiation of the combined effect of hyperthermia and chemotherapy with ADR may depend only upon the dose in one of these two modalities.     

Results in combined hyperthermia/radiotherapy of adenoid cystic carcinomas compared with conventional treatment

158 patients with advanced malignancies of the head and neck area were treated by a combined hyperthermia/radiotherapy since 1972. 16 were adenoid cystic carcinoma (ACC). Their treatment results are compared with 28 ACC after radiotherapy alone. All 16 patients with hyperthermia achieved a complete remission. CR in 28 ACC-patients without hyperthermia was 61%. 113 patients with hyperthermia-treated squamous cell carcinomas had a comparatively low CR-rate of 49%. Five-year-survival on the other hand was 50% for ACC after hyperthermia, 57% after a planned postoperative radiation without hyperthermia and 37.5% after a radiotherapy alone in cases of inoperability. The combination of hyperthermia and radiotherapy is indicated in cases of advanced inoperable ACC and leads to a high percentage of local tumour control.     

加热复合电离辐射对人骨髓细胞的影响

目的：研究了加热与电离辐射后对人离体骨髓细胞存活率及形态等方面的影响。结果：骨髓细胞存活率随加热时间增加而呈直线下降，温度越高直线斜率越大。在形态学上，骨髓细胞受热后可出现非特异性退行性变化，随着时间的延长，这些变化更加明显。骨髓细胞受单纯Ｘ射线照射后，可见随着照射剂量的增大，细胞存活率明显下降。形态学上单纯３ＧｙＸ射线照射可使少数细胞出现空泡变性，线粒体肿胀，核膜破裂等变化。骨髓细胞受Ｘ射线照射后立即给予４２℃３０分钟的加热使骨髓细胞存活率下降的程度比各自单独作用之和要大（Ｐ＜０．０１）。电镜显示结构破坏明显，细胞支架系统、细胞器均受到破坏，细胞膜、核膜破坏乃至溶解，核浆、细胞浆外溢。     

Deoxyribonucleic acid synthesis by rat thymus and spleen cells in vitro following hyperthermia

The inhibition of the semiconservative and restorative DNA synthesis caused by hyperthermia (30 to 60 min, 43 degrees C) was significantly higher in spleen cells than in thymus cells. The DNA repair synthesis of thymus cells measured at 37 degrees C was increased by about two times the initial value after a pre-incubation of 30 to 90 min and 30 to 60 min, respectively, with 37 and 43 degrees C, respectively. Under the same conditions, the 3H-thymidine incorporation into the DNA of spleen cells diminished proportionally to the pre-incubation time after a pre-incubation of 30 and 45 min, respectively, with 43 and 37 degrees C, respectively. When hyperthermia and inhibitors of DNA synthesis or DNA repair (hydroxyurea, 1-beta-D-arabinofuranosylcytosine, 3',5'-didesoxythymidine, and 3-aminobenzamide) were combined, overadditive effects--without cell specific particularities--were seen only in the case of 3-aminobenzamide. Only in thymus cells, the inhibitor of DNA topoisomerase II novobiocin caused an overadditive reinforcement of the inhibition induced by hyperthermia of the semiconservative DNA synthesis. The stimulation of DNA repair synthesis in thymus cells caused by novobiocin with the aid of DNA polymerase beta could be compensated by hyperthermia. The sedimentation of thymus and spleen cell nucleoids was increased after hyperthermia. The results suggest a special importance of DNA topology and of the DNA polymerase beta activity for the cellular effect of hyperthermia.     

Radiation and thermal characteristics of mouse lymphoma cells and their radiation-sensitive mutant

Radiation and thermal characteristics of L5178Y cells and their radiation-sensitive mutant M10 cells were studied by the colony-forming method and the dye-exclusion method using eosin-Y. Although M10 cells were remarkably radiation-sensitive compared with L5178Y cells, it was difficult to cause interphase death of M10 after a large dose of irradiation. After heat treatments, L5178Y cells revealed more cell destruction and were stained well by eosin-Y, but it was relatively difficult to produce cell destruction of M10 cells, which showed poor staining by eosin-Y. When assayed by the colony-forming method, M10 cells were also heat-resistant compared to L5178Y. The dye-exclusion rate was closely correlated with cell survival after hyperthermia of L5178Y cells, suggesting that this is a simple method of detecting the thermosensitivity and thermotolerance of cancer cells. The difference in survival of L5178Y cells and M10 cells after combined treatment with gamma irradiation and hyperthermia was smaller than with gamma irradiation alone. It was also found that there was a relationship between radiation-induced interphase death and hyperthermia-induced interphase death, and that interphase death accounted for a major part of cell death caused by hyperthermia in mouse leukemia cells.     

恒温循环热灌注化疗治疗恶性胸腔积液临床研究

 目的 观察恒温循环热灌注化疗对恶性胸腔积液的治疗效果、治疗后患者生活质量的改善及其不良反应.方法 将我科确诊恶性胸腔积液患者180例,随机分为治疗组(90例)和对照组(90例),治疗组行胸腔恒温循环热灌注化疗,对照组仅行胸腔热灌注化疗.两组均胸腔内注入顺铂(DDP),首剂90 mg,后每次60 mg,每周1次,共6次.结果 治疗组控制胸水有效率(CR+PR)为94.4%,对照组为64.4%,两组间比较差异有统计学意义(P<0.01).治疗组与对照组生活质量好转率分别为85.56%和62.22%( P<0.01).不良反应方面,消化道反应与血液学毒性,两组间差异均无统计学意义 (P=1.000).结论 采用胸腔恒温循环热灌注化疗方法治疗恶性胸腔积液疗效确切,安全性高,值得在临床广泛推广.     

Comparative evaluation of combined irradiation and hyperthermia versus irradiation alone

Controvesy remains as to the treatment schedule producing better results in combined hyperthermia and X-ray therapy. Our experience concerning combined therapy of the solid tumour Walker carcinoma is reported. Male Wistar rats were submitted to treatment on the ninth day after transplantation of the tumour. Two groups of rats received either a therapeutic X-ray dose of 800 cGy by a 6-MeV linear accelerator (Mevatron, Siemens) or treatment by 432 MHz of microwaves with continuous control of tumour tissue temperature to 44 ± 1 °C for 45 min. Another group of rats was submitted to a combined treatment, with X-ray therapy preceding hyperthermia by 24 h. The last group of animals constituted the control rats. Greater tumour regression and longer survival times were obtained with the combined treatment. The gain factor for survival time was equal to 1.85 after combined treatment compared with 1.30 after X-ray therapy and 1.05 after hyperthermia. In conclusion, the results suggest that in the above schedule of combined treatment, hyperthermia applied to a solid tumour 24 h after a single dose of X-rays enhances the beneficial effect of therapy. Correspondence to: C. Sawas-Dimopoulou     

Histological and morphometric research on rhabdomyosarcoma in the rat after irradiation and microwave hyperthermia

Cellular effects of a combined treatment with X-rays (15 Gy) and local microwave hyperthermia (43 degrees C, 60 min) on the tumor tissue of the rhabdomyosarcoma R1H of the rat were studied with light and electron microscopy. The results were compared with the cell reactions obtained after X-rays (15 Gy) alone. The histological results and the morphometric data show that the time course of cell damage changes individually with the treatment applied. After radiation alone a cell oedema occurs within some days. After a combined treatment, however, a marked cell oedema and cell lysis is already observed after 24 hours. An intensive shrinking of the cell nuclei is found at day 6th, attended with pyknosis, karyolysis and cell fragments in the extended extracellular area. In comparison with radiation alone the processes of repair and repopulation in the R1H tissue are remarkably delayed after a combined treatment.     

综合治疗原发性肝癌的临床观察

目的分析伽玛刀配合热疗治疗原发性肝癌的效果。方法自2004年4月-2005年8月,原发性肝癌患者65例行单纯伽玛刀治疗,31例给予伽玛刀治疗配合热疗。结果治疗后3个月总的有效率为83.3%（80/96）。单纯伽玛刀组的1年、2年局部控制率分别为61.5%（40/65）、35.4%（23/65）,1年、2年生存率分别为63.1%、32.3%;伽玛刀配合热疗组的1年、2年局部控制率分别为64.5%（20/31）、35.5%（11/31）,1年、2年生存率分别为67.7%,38.7%。随访期内未见严重放射性并发症。结论对原发性肝癌采用伽玛刀配合热疗是较有效的局部治疗方式。