热化疗对晚期恶性肿瘤患者外周血CD8+CD28+ T细胞表达的影响及临床意义

 目的探讨热化疗对晚期肺癌、大肠癌、乳腺癌患者外周血CD8+CD28+T细胞亚群（CTL）表达的影响及临床意 义。方法晚期肺癌、乳腺癌、大肠癌患者各20例,随机分为热化疗与化疗两组,每组肺癌、乳腺癌、大肠癌患者各 10例,热化疗组每周期化疗予以同步全身热疗,至少完成两个周期治疗,正常健康志愿者20例作为健康对照,用流 式细胞仪检测外周血CD4+、CD8+、CD8+CD28+T细胞亚群。结果与健康对照组相比,肿瘤患者外周血T细胞CD4+ 、 CD8+及CD4+/CD8+的表达差异无统计学意义(P＞0.05),CD8+CD28+的表达显著降低(P＜0.05),热化疗组患者治疗后 T细胞CD8+CD28+表达上调,与治疗前比较差异有统计学意义（P＜0.05）,化疗组患者治疗前后T细胞CD8+CD28 +的 表达差异无统计学意义（P＞0.05）,热化疗组患者的有效率、生存质量优于化疗组患者（P＜0.05）, 两组治疗 的Ⅲ~Ⅳ度骨髓抑制及消化道反应比较差异无统计学意义（P＞0.05）,T细胞CD8+CD28+的表达与肿瘤病理类型无关 。结论晚期肿瘤患者T细胞存在共刺激分子CD28表达的低下及肿瘤细胞的免疫逃逸,热化疗能上调T细胞CD8+CD28+ 的表达,增强肿瘤患者的免疫监视功能,在疗效与预后上较化疗有优势。     

结肠癌患者化疗/热化疗前后CD4+CD25+T细胞和CTL变化

背景与目的:化疗是内科治疗结肠癌的主要方法之一,热化疗改善患者机体免疫状态的机制尚不清楚.本研究通过观察结肠癌患者化疗/热化疗前后CD4+CD25+调节性T(Treg)细胞及细胞毒T淋巴细胞功能的变化,探讨其临床意义.方法:入选结肠癌Ⅲ期术后患者60例,其中热化疗组30例,以单纯化疗组30例为对照组,收集患者治疗前后外周血,采用流式细胞术检测治疗前后外周血淋巴细胞中CD4+CD25+Treg细胞的表达,并检测患者CTL细胞杀伤功能.结果:(1)结肠癌患者单纯化疗组化疗前CD4+CD25+Treg细胞表达为19±9,化疗后为19±7,差异无统计学意义(F=0.026,P＞0.05);热化疗组化疗前CD4+CD25+Treg细胞表达为19±5,化疗后为14±4,差异有统计学意义(F=2.198,P＜0.05);(2)结肠癌患者单纯化疗组化疗前细胞毒T淋巴细胞(cytotoxic T lymphocytes,CTL)为10±3,化疗后为15±5,差异有统计学意义(F=187.801,P＜0.05);热化疗组化疗前CTL为12±3,化疗后CTL为18±5,两组比较差异有统计学意义(F=236.097,P＜0.05).即化疗组治疗前后CD4+CD25+Treg细胞变化不明显,而化疗联合热疗组治疗后CD4+CD25+Treg细胞水平显著降低.两组结肠癌患者治疗后CTL细胞杀伤功能均明显增强,以热化疗组增强更明显.结论:热化疗可明显改善结肠癌患者的免疫抑制状态,增强患者的细胞毒T淋巴细胞的功能.化疗联合热疗可提高患者疗效和预后.     

热化疗对荷瘤鼠NO水平影响的研究

目的:观察热化疗对荷瘤鼠NO水平的影响。方法:80只荷瘤Balb/c小白鼠随机分为不治疗组(NT),化疗组(P),热疗组(H)和热化疗组(HP),每组20只(雌雄各半)。处理后观察其抑瘤情况及肿瘤组织形态学变化,并采用硝酸还原酶法检测瘤组织内NO的浓度。结果:热化疗组与热疗组,热化疗组与化疗组抑瘤结果有显著性差异(P<0.01)。除热疗组和化疗组之间差异无统计学意义(P>0.05)外,其余各组之间NO含量均有显著性差异(P<0.01)。结论:热化疗具有良好的抗肿瘤效应,可以显著提高荷瘤宿主NO水平,荷瘤宿主血清NO具有双重作用,低浓度时可促进肿瘤生长,高浓度时参与抗肿瘤免疫、抑制和杀伤肿瘤细胞。     

热化疗与热疗治疗晚期肺癌32例临床观察

 目的　比较热化疗、热疗对晚期肺癌的近期疗效及其对患者外周血白细胞总数、CEA、TSGF水平的影响。方法　热化组 17例 ,热疗组 15例 ,观察其近期疗效 ,检测其治疗前后外周血白细胞总数、CEA、TSGF水平。结果　热化组、热疗组有效率分别为 70 .5 %、4 6 .6 % ,两者比较无显著性差异 (P >0 .0 5 ) ,两组血清CEA、TSGF水平治疗后均下降且有显著性差异 (P <0 .0 5 ) ,外周血白细胞总数治疗前后均无显著性差异 (P >0 .0 5 )。结论　热化疗较热疗治疗晚期肺癌有更好的疗效 ,热疗与化疗相结合能防治白细胞减少     

热化疗对荷瘤鼠IL-2、TNF水平影响的研究

目的：探讨热化疗对荷瘤鼠IL-2、TNF水平的影响。方法：对正常Balb/c鼠及实验组荷瘤鼠脾脏IL-2、TNF水平进行检测。结果：热化疗组荷瘤鼠IL-2、TNF水平与正常鼠无显著性差异(P＞0.01)，而与单纯热疗或单纯化疗及未作任何处理组比较有显著性差异(P＜0.01)。结论：热化疗可以提高荷瘤鼠IL-2、TNF水平，从而增强机体的免疫功能，提高机体全身抗肿瘤效应。     

热化疗对荷瘤鼠IL—2、TNF水平影响的研究

目的：探讨热化疗对荷瘤鼠IL－2、TNF水平的影响。方法：对正常Balb／c鼠及实验组荷瘤鼠脾脏IL－2、TNF水平进行检测。结果：热化疗组荷瘤鼠IL－2、TNF水平与正常鼠无显著性差异（P＞0．01），而与单纯热疗或单纯化疗及未作任何处理组比较有显著性差异(P＜0．01）。结论：热化疗可以提高荷瘤鼠IL－2、TNF水平，从而增强机体的免疫功能，提高机体全身抗肿瘤效应。     

热疗配合腹腔灌注化疗治疗恶性腹腔积液的临床观察

目的:探讨热疗联合腹腔灌注化疗治疗恶性腹腔积液的临床疗效和不良反应。方法:将恶性腹腔积液患者58例分为两组,腹腔灌注化疗同时配合热疗组(治疗组)30例,单纯腹腔灌注化疗组(对照组)28例。评估疗效及不良反应。结果:腹腔积液疗效评价:治疗组有效率为86.7%(26/30),对照组有效率57.2%(16/28),两组差异有统计学意义(P=0.012);生活质量评价:治疗组改善96.7%(29/30),对照组改善71.4%(20/28),两组差异有统计学意义(P=0.008)。两组的不良反应发生率差异无统计学意义(P>0.05)。结论:热疗加局部腹腔灌注化疗治疗恶性腹腔积液疗效肯定,不良反应轻微,值得临床推广。     

热疗联合化疗治疗晚期非小细胞肺癌临床研究

目的:比较热疗联合化疗治疗晚期非小细胞肺癌(NSCLC)的疗效及安全性。方法:将225例Ⅲb/Ⅳ病人1∶2∶4比例随机分为全身热疗联合化疗组、局部热疗联合化疗组、单纯化疗组。全身热疗采用SRI全身热疗系统,每周期1次;局部热疗采用HG-2000高频热疗机,每周期2次;化疗采用含铂两药联合方案。结果:无进展生存期(PFS):全身热疗组35例,(5.21±3.84)月;局部热疗组61例,(4.02±2.82)月;单纯化疗组129例,(3.49±2.75)月。全身热疗组PFS与单纯化疗组对比有显著统计学差异(P=0.003),也优于局部热疗联合化疗组(P=0.061)。局部热疗联合化疗组与单纯化疗组相比无显著统计学差异(P=0.255)。治疗相关毒副反应各组之间无统计学差异。结论:全身热疗联合化疗PFS优于单纯化疗及局部热疗联合化疗组,毒副反应可耐受,有望成为晚期NSCLC的辅助治疗方法。     

鼻咽癌组织中HSP70和Ki-67的表达及其对预后影响的评价

目的探讨热休克蛋白70(heat shockprotein70,HSP70)和Ki-67在鼻咽癌组织中的表达及与预后的关系。方法采用免疫组织化学二步法,观察50例鼻咽癌患者鼻咽部或颈淋巴结活体组织检查存档石蜡标本中HSP70和Ki-67蛋白的表达。结果50例鼻咽癌组织中HSP70和Ki-67阳性表达率分别为36.0%(18/50)和98.0%(49/50);HSP70在T分期中的阳性表达差异无统计学意义;HSP70(+)组生存率(10.9%)与HSP70(-)组(33.8%)相比,差异有统计学意义,χ2=4.10,P=0.043;Cox模型多因素分析显示,HSP70(+)表达是独立的预后不良因素之一,P=0.031。在临床研究指标(T分期、N分期、生存率等)中,Ki-67的高表达(++/+++)率与低表达(+/-)率比较均差异无统计学意义。结论HSP70表达可作为评价鼻咽癌患者预后的指标。     

亚高温盆腔区域热疗对原位大鼠膀胱肿瘤形态学影响的研究

 目的 研究亚高温盆腔区域热疗致大鼠膀胱肿瘤形态学改变。方法 制备原位的大鼠膀胱肿瘤动物模型，用内生场热疗系统从体外加热大鼠盆腔至41 ℃，热疗后对肿瘤进行病理研究。结果电镜观察短期组有早期凋亡细胞出现；长期组凋亡细胞和凋亡小体多见。肿瘤细胞HSP70表达率短期组中热疗组均高于对照组，差异有统计学意义（P＜0.01），而长期组中差异无统计学意义。PCNA LI热疗组均低于对照组，差异有统计学意义（P＜0.01）。AI热疗组均高于对照组，差异有统计学意义（P＜0.01）。结论亚高温盆腔区域热疗抑制肿瘤细胞增生，诱导凋亡，长期热疗未导致热耐受。     

口腔鳞癌组织中HSP70基因mRNA的表达及其意义

目的检测HSP70基因的信使RNA在不同分化口腔鳞癌中的表达,以探讨其在口腔鳞状细胞癌组织中表达的意义。方法用半定量RT-PCR方法检测30例口腔鳞状细胞癌和20例正常口腔黏膜中HSP70基因mRNA的表达,并分析HSP70基因mRNA表达量与临床病理特征的关系。结果 HSP70基因mRNA在高、中、低分化口腔鳞状细胞癌及正常组织中的灰度平均值为1.07±0.04,1.21±0.05,1.36±0.03;正常口腔黏膜上皮HSP70 mRNA的灰度平均值为0.95±0.15,表达低于口腔鳞状细胞癌组织,两者之间的差异具有显著性,高中低分化的口腔鳞状细胞癌两两比较,均有统计学差异。结论 HSP70基因表达与口腔鳞状细胞癌发生发展呈正相关关系,这对诊断口腔鳞状细胞癌有重要意义。但是HSP70的mRNA低表达则不能完全当作是口腔鳞状细胞癌中的早期事件。     

Expression of 70-kDa heat shock protein in oral lesions: marker of biological stress or pathogenicity

We showed differential expression of HSP70 during oral tumorigenesis. The precise functional role of HSP70 overexpression in the pathogenesis of betel and tobacco related oral cancer remains to be determined. To evaluate the utility of HSP70 as an indicator of the biological stress experienced by tumour cells or the malignant potential of oral epithelial lesions and predicting clinical outcome, its expression was assessed in different stages of oral carcinogenesis by immunohistochemical analysis and correlated with clinicopathological parameters. Overexpression of HSP70 protein was observed in 38 of 64 (59%) dysplastic lesions and 92 of 125 (74%) oral squamous cell carcinomas (SCCs) which included 76 of 105 cases (72%) of primary oral SCCs and 16 of 20 (80%) of recurrent oral SCCs. A significant correlation of HSP70 expression was observed with severity of dysplasia (P=0.0006767), poor histological differentiation of primary tumours (P=0.0184348), increase primary tumour size (P=0.0221103) and consumption of betel and tobacco (P<0.01). Follow-up studies showed that in patients with premalignant lesions the median transition time (premalignancy to malignancy) was significantly shorter in HSP70 overexpressing cases than those showing basal level of HSP70 (P=0.012). Oral cancer patients with elevated levels of HSP70 showed decreased median disease-free survival time (no recurrence/metastasis) than those showing basal HSP70 immunoreactivity (P=0.0246). The results suggest that HSP70 expression may not be a mere marker of biological stress but may also be implicated in the pathogenesis of oral cancer.     

Induction of apoptosis by abrogation of HSP70 expression in human oral cancer cells

We have reported differential expression of 70-kDa heat shock protein (HSP70) in human oral tumorigenesis. The functional significance of elevated levels of HSP70 protein in oral squamous cell carcinomas (SCC) remains to be elucidated. The present study was designed to investigate the role of HSP70 protein in the proliferation and survival of oral tumour cells. Abrogation of HSP70 expression by antisense HSP70 oligonucleotides treatment of human oral carcinoma cells isolated from primary tumours or HSC-2 cells triggered cell death with several characteristic features, including DNA laddering, chromatin condensation and fragmentation. Flow-cytometric analysis showed a hypodiploid DNA peak of propidium iodide-stained nuclei in the antisense oligomer-treated cells. This response was accompanied by a decrease in the percentage of cells in the S phase of the cell cycle, suggesting inhibition of cell proliferation. Treatment of oral cancer cells with HSP70 antisense oligomers resulted in decreased expression of anti-apoptotic signal protein bcl-2. Our results suggest that HSP70 antisense oligomer treatment abrogates the expression of HSP70 protein that may disrupt HSP70-bcl-2 interactions, in turn inhibiting cell proliferation and inducing apoptosis. Conversely, the data suggest that HSP70 is required for proliferation and survival of oral tumour cells.     

HSP70 as a Diagnostic and Prognostic Marker in Egyptian Women With Breast Cancer

BackgroundHeat shock protein 70 (HSP70) is a significant cellular stress response protein that has intrinsic and extrinsic pathways to protect cells against apoptosis. It is one of the most induced proteins in cancer cells. The aim of the present study is to investigate the significant role of the HSP70 expression in Egyptian patients with breast cancer (BC) and its potential to be as a diagnostic and prognostic marker.Materials and MethodsHSP70 was examined in 155 cases in this prospective study; patients were subdivided into 3 groups: 60 patients with malignant metastatic disease, 60 patients with malignant non-metastatic disease, and 35 patients with benign lesions as control. HSP70 expression was detected using enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry (IHC).ResultsMost cases of breast cancer expressed HSP70 in both serum (98.3%) and tumor tissue (90%). A strong positive correlation was found between HSP70 IHC and ELISA (r = 0.811). The mean HSP70 levels, as detected in both patients’ serum by ELISA and tumor tissue by IHC, was significantly higher in patients with BC than in benign cases (P = .001). HSP70 was significantly higher in patients with metastatic BC than in those with non-metastatic BC (P = .001). HSP70 showed positive correlation with tumor size (pT stage) and number of lymph node metastases (P ≤ .001).ConclusionHSP70 is over-expressed in patients with metastatic and non-metastatic BC than in benign cases. A high level of HSP70 either in patient’s serum or in tumor tissue correlated significantly with advanced disease in patients with BC. This present study suggests that HSP70 can serve as a BC biomarker for early screening, diagnosis, and follow-up.     

Anti-Tumor Effect of Heat Shock Protein 70-Peptide Complexes on A-549 Cells

Objective: To investigate the anti-tumor immunity in vitro of heat shock protein 70-peptide complexes (HSP70-PC) from human lung cancer tissue. Methods: HSP70-PC was purified from lung tumor tissues and corresponding non-tumor lung samples with the methods of ADP-affinity chromatography, DEAE ion-exchange chromatography and Western-blot. The activation and proliferation of PBMC induced by different HSP70-PC and tumor cytotoxic reactivity to A549 cells in vitro were measured by the MTT cell proliferation assay. Results: The purified HSP70-PC had a very high purity found by SDS-PAGE and Western-blot. Human lymphocytes were sensitized efficiently by HSP70 preparation purified from lung cancer tissues and a definite cytotoxicity to A-549 cells was observed. There was significant difference with HSP70-PC purified from lung cancer, compared with the control group (P<0.001). Conclusion: High purity of HSP70-PC could be achieved from tumor tissues in this study. HSP70-PC purified from human tumor tissues can induce anti-tumor immunity in vitro mainly implemented by eliciting CTL immunity.     

主要人热休克蛋白在癌组织及癌旁正常组织中表达水平的比较研究

目的对２５例喉癌及癌旁正常喉组织中几种主要人热休克蛋白的ｍＲＮＡ表达水平进行检测和比较。方法采用ｍＲＮＡ狭缝杂交的方法。ＨＳＰ２７、ＨＳＣ７０和ＨＳＰ９０β探针用ＰＣＲ方法进行制备。结果ＨＳＰ９０α和ＨＳＰ７０在喉癌组织中的表达量显著高于正常喉粘膜,平均表达量均为正常喉粘膜表达量的５倍。而ＨＳＰ２７、ＨＳＣ７０和ＨＳＰ９０β的表达量,在两种组织中变化不大。结论与其他热休克蛋白相比,ＨＳＰ７０和ＨＳＰ９０α可能在喉癌的发生过程中有更为重要的作用。进一步深入研究它们在肿瘤和正常组织中不同表达的机理,将为热休克蛋白用于喉癌的生物治疗提供理论依据。     

来源于人膀胱癌细胞株EJ的HSP70负载的DC所激发的特异性CTL细胞的体外应答效应

Objective: To investigate whether human dendritic cells (DC) derived from peripheral blood mononuclear cells (PBMC), which were pulsed by heat shock protein 70 (HSP70) isolated from human bladder tumor cell lines of EJ, were able to induce peptide specific cytotoxic T-lymphocytes (CTL) response in vitro and give the experimental foundation for the future clinical trials of immunotherapy in bladder tumor. Methods: The EJ-derived HSP70 co-cultured with DC from the healthy volunteers' PBMC, along with the crude lysate (the supernatant before HSP70 purification) from EJ cells were used as the experimental groups and DC not pulsed by any tumor cells antigen were the blank control. The autologous T-lymphocytes were added into the above various DC groups, and after incubation, the stimulation indexes (SI) and interferon-γ (IFN-γ) were detected to evaluate the immune activities of various DC groups. The killing effects of CTL to target cells, EJ and Hela cells, were determined with 51Cr releasing test. Results: Both DC/HSP70 and DC/the crude lysate could effectively activate CTL in vitro and kill target cells EJ. The killing effect of DC/HSP70 to EJ was much stronger than DC/the crude lysate (the supernatant before HSP70 purification) (P < 0.05). DC without any tumor cell antigens had a lower killing power to EJ. Meanwhile, DC/ HSP70 had little killing power to Hela non-relevant to bladder tumor histopathologically as compared with EJ cells (P < 0.05). Conclusion: The DC pulsed by HSP70 derived from the autologous tumor cells could induce a peptide complexes specific CTL response to tumor cells, and the CTL response induced by the DC/HSP70 was stronger, which display the basis of the possible clinical application of DC/HSP70 for bladder tumor.     

Ｓｕｒｖｉｖｉｎ、ＨＳＰ７０和Ｆａｓ在乳腺癌中的表达及其意义

目的　探讨人乳腺癌组织、癌旁正常乳腺组织中Ｓｕｒｖｉｖｉｎ、ＨＳＰ７０和Ｆａｓ的表达及其临床意义。方法　应用免疫组化法检测３６例患者的乳腺癌组织及癌旁正常乳腺组织标本中Ｓｕｒｖｉｖｉｎ、ＨＳＰ７０和Ｆａｓ蛋白的表达情况。结果　Ｓｕｒｖｉｖｉｎ、ＨＳＰ７０和Ｆａｓ在乳腺癌组织中的阳性表达率分别为８３.３％、８６.１％和５２.８％,在癌旁组织中表达率分别为４１.７％、６３.９％和９７.２％。Ｓｕｒｖｉｖｉｎ、ＨＳＰ７０在乳腺癌组织中的阳性表达率明显高于癌旁正常乳腺组织,而Ｆａｓ在乳腺癌组织中的阳性表达率明显低于癌旁正常乳腺组织。Ｓｕｒｖｉｖｉｎ、ＨＳＰ７０和Ｆａｓ的表达与年龄、病理类型、肿瘤直径、ＨＥＲ-２无显著性差异（Ｐ＞０.０５）;Ｓｕｒｖｉｖｉｎ和ＨＳＰ７０的表达率在有淋巴结转移组明显高于无淋巴结转移组（Ｐ＜０.０５）,而Ｆａｓ的表达率在有淋巴结转移组明显低于无淋巴结转移组（Ｐ＜０.０５）;且Ｓｕｒｖｉｖｉｎ在ＥＲ（－）／ＰＲ（－）组表达率高（Ｐ＜０.０５）。Ｓｕｒｖｉｖｉｎ与Ｆａｓ表达呈负相关（ｒ＝－０.４２３,Ｐ＜０.０５）;Ｓｕｒｖｉｖｉｎ与ＨＳＰ７０表达无明显相关性（ｒ＝０.２５１,Ｐ＞０.０５）。结论　Ｓｕｒｖｉｖｉｎ、ＨＳＰ７０上调及Ｆａｓ下调可能在乳腺癌发生发展中起着一定的作用。     

热休克蛋白70在乳腺癌中的表达及其与预后的关系

目的 探讨热休克蛋白70(heat shock protein70，HSP70)在乳腺癌中的表达，评估其在乳腺癌预后中的作用。方法 用免疫组化方法检测62例手术切除的乳腺癌组织中HSP70的表达，研究其与临床病理特征的关系及对预后的影响。结果 (1)HSP70在乳腺癌组织中的阳性表达率为56．5％(35／62)。(2)HSP70表达与肿瘤大小(P=0．021)及腋窝淋巴结转移(P=0．015)存在相关性，而与月经状况、组织学分级之间均无相关性。(3)HSP70阴性表达组的无病生存期(DFS)及总生存期(OS)均明显优于阳性表达组。(4)(20x回归单因素分析显示腋窝淋巴结转移个数、组织学分级、肿瘤大小、TNM分期及HSP70阳性表达与DFS及OS明显相关；Cox回归多因素分析表明仅有腋窝淋巴结转移个数、组织学分级及肿瘤大小与DFS及OS明显相关，而TNM分期及HSP70阳性表达进入Cox回归模型。结论 HSP70在乳腺癌组织中有一定程度的表达，HSP70阳性表达与乳腺癌患者生存期的缩短有关，由于多因素回归分析未能显示HSP70表达与DFS和OS相关，因此HSP70不能作为乳腺癌的独立的预后因素。     

Overexpression of the heat shock protein HSP70 family and p53 protein and prognosis for patients with gastric cancer

The cell synthesis of heat shock proteins is increased by a variety of environmental and pathophysiological stressful conditions. The 70-kD heat shock protein family (HSP70 family) which constitutively expresses hsc70 and heat-inducible hsp70 is thought to be involved in protein-protein interactions, including oncogene products. We investigated the HSP70 family expression and biological behavior of gastric cancer, and its relation to p53 overexpression. Expressions of HSP70 and p53 in 164 primary gastric tumors were determined immunohistochemically. Exploratory data were analyzed on a set of 164 primary gastric cancers, and we constructed in prognostic significance of the HSP70 expression level and the relation to p53 overexpression. Expression of HSP70 (hsc70 and hsp70) were detected in nuclei and/or cytoplasm of cancer cells. Western blotting analysis showed that hsc70 and hsp70 were both expressed in five gastric cancer cell lines. Immunohistochemically stained positive cells of HSP70 varied from 0 (very weak) to 100%, in each case. The median level of positive cell rate was 19.0%. A HSP70 expression of over 19.0% was related to the differentiated tissue type of gastric cancer, but not to other clinicopathological factors. There was no difference in survival rates in subjects with higher and lower groups of HSP70 expression. HSP70 expression was also not related to p53 overexpression in the nuclei and p53 overexpression-related poorer prognosis. Our findings show that the expression of HSP70 is not associated with tumor advance-related characteristics or with the prognosis of gastric cancer. Measurements of HSP70 expression do not appear to be a useful prognostic marker.