食管、贲门

食管癌高发区上消化道恶性肿瘤死亡趋势分析,食管癌高发区队列内镜筛查随访分析,河南林州食管癌高发区反流性食管炎调查,食管癌转移淋巴结与原发肿瘤EGFR和c-erbB-2的表达,人胎儿发育过程中食管上皮组织p21和PCNA蛋白的表达,     

食管癌中P16、PCNA基因蛋白的表达及意义

 目的：探讨P16、PCNA基因蛋白表达与食管鳞癌发生发展的关系。方法：应用S-P免疫组织化学方法对84例食管鳞癌及10例正常食管组织分别检测P16蛋白、PCNA蛋白表达。结果：84例食管鳞癌中P16蛋白表达率为54.8%(47/84),P16表达与淋巴结转移、临床分期、术后生存期显著相关(P<0.05);PCNA表达阳性率与食管癌组织分化程度显著性正相关(P<0.05),淋巴结转移组显著高于未转移组,并与术后生存期互相关(P<0.01),与临床分期无显著相关。P16与PCNA表达呈反相关系(P<0.01)。结论：表明P16在食管癌生长、转移中起重要作用,P16、PCNA可作为判断预后的重要指标。     

p53及PCNA的异常表达在食管上皮增生和癌变过程中的意义

目的:研究食管上皮增生、不典型增生及原位癌中p53蛋白及增殖细胞核抗原(PCNA)的异常表达,探讨其在食管癌发生发展中的作用.方法:用免疫组化LSAB方法检测189例食管鳞癌癌旁上皮及原位癌中p53及PCNA的表达.结果:癌旁上皮中存在p53蛋白积聚,从上皮增生→不典型增生→原位癌,其阳性率依次为55 %、79 %和98 %(P＜0.01).PCNA的阳性强度也是依次递增.结论:p53蛋白的积聚在食管鳞癌癌前病变中既已存在,说明它是一个早期事件,在食管癌的发生中起一定的作用.p53及PCNA的异常表达可能成为判断食管上皮发生癌变或癌前病变的客观指标之一.     

食管鳞状细胞癌中nm23-H1和p53蛋白表达及其相互关系

目的:探讨食管鳞癌组织中p53和nm23-H1蛋白的表达与癌组织分化浸润转移的关系,以及探讨两者之间的相关性,并进一步分析癌组织中p53和nm23-H1蛋白表达对食管癌患者的预后意义。方法:采用免疫组织化学(S-P法)方法对100例人食管鳞癌组织中的p53和nm23-H1蛋白的表达情况进行检测。结果:100例食管鳞癌组织中,nm23-H1阳性表达者70例(阳性率为70%),p53阳性表达者64例(阳性率为64%)。nm23-H1蛋白表达与食管癌淋巴结转移有关(P<0.025),与食管鳞状细胞癌的分化程度、肿瘤部位、浸润深度、病变长度以及患者性别、年龄无关(P>0.05)。p53蛋白表达与食管鳞状细胞癌的分化程度、浸润深度有关(P<0.05),与食管癌淋巴结转移、肿瘤部位、病变长度、患者性别、年龄无关(P>0.05)。高分化鳞癌组织中p53明显低表达(29.2%);低分化鳞状细胞癌组织中p53表达明显增高(71.4%)。食管外膜受累者p53表达较高(56%);仅发生食管粘膜和(或)粘膜下浸润组的癌组织中未发现有p53蛋白的表达。食管癌组织中nm23-H1蛋白低(高)表达与p53高(低)表达之间有明显相关性(P<0.01)。nm23-H1和p53蛋白表达亦与食管癌的TNM分期密切相关(P<0.05)。食管癌TNM分期越晚,其癌组织中nm23-H1蛋白表达越低,p53蛋白表达越高。结论:nm23-H1基因低表达与p53基因高表达可能在食管鳞状细胞癌浸润转移过程中发挥重要作用。nm23-H1可以作为食管鳞状细胞癌患者预后的基因标记,其蛋白表达产物的检测可以用于患者预后的判断,并为患者治疗方案的制定提供参考。     

抑癌基因p53在不同地区食管癌中的表达及临床预后分析

目的：比较中国食管癌高，低发区p53蛋白的积聚，燕对p53蛋白积聚与肿瘤细胞增殖及临床预后的关系进行探讨。方法：用免疫组化LSAB法检测高发区（河南）43例食管癌和低发区（广东）40例食管癌p53蛋白及增殖细胞核抗原的表达。结果：p53蛋白阳性率分别是60．5％（河南）和57．5％（广东）。阳性率差异无显著性（P＞0．05）。p53蛋白表达无论在高，低发区均与PCNA的表达密切相关（P＜0．001）。结论：食管癌不同的高，低发区中p53蛋白的各聚是类同的。大多数p53蛋白积聚的肿瘤细胞是处于细胞增殖周期内，可能成为食管癌恶性程度的指标之一，将为临床分析预后提供客观依据。     

食管鳞癌中p53、PCNA和EGFR的表达与化疗疗效的关系

目的:探讨p53、PCNA和EGFR在食管鳞癌中的表达情况与化疗疗效的关系。方法:对66例食管鳞癌化疗前的活检标本用免疫组化技术分别检测p53、PCNA、EGFR的表达。结果:p53蛋白积聚阳性组化疗有效率(16.7%)明显低于p53蛋白积聚阴性组(70.8%)(P<0.01),PCNA过表达组化疗有效率(79.4%)高于PCNA弱表达组(31.3%)(P<0.01),EGFR过表达组化疗有效率(30.4%)低于弱表达组(69.8%)(P<0.01)。经Logistic回归分析,3个指标中,PCNA对化疗疗效的预测价值较大(P<0.01)。结论:食管鳞癌中p53、PCNA和EGFR的表达情况对化疗疗效均有一定的预测价值。其中PCNA价值较大。     

PTEN在食管鳞状细胞癌中的表达和临床意义

目的：研究抑癌基因PTEN蛋白表达与食管鳞状细胞癌临床病理特征的关系,探讨其在食管癌变中的可能作用。方法：应用免疫组织化学SP法检测60例食管鳞状细胞癌及20例癌旁正常组织中PTEN蛋白的表达,结合临床资料进行分析。结果：PTEN蛋白阳性反应主要定位于胞浆。食管鳞状细胞癌组织中PTEN蛋白表达阳性率56．7％明显低于正常组织阳性率90．0％（P〈0．05）。PTEN蛋白在低分化、中分化、高分化鳞癌组的阳性表达率分别是21．4％、55．0％、76．9％,三组之间相互比较有极显著统计学差异（P〈0．01）,肿瘤分化程度越低PTEN蛋白表达越低。有淋巴细胞转移的食管癌组织中VrEN表达的阳性率42．9％明显低于无淋巴细胞转移的食管癌组织阳性率76．0％（P〈0．05）。有外膜浸润的食管癌组织中PTEN表达的阳性率26．1％明显低于无外膜浸润的食管癌组织阳性率75．7％（P〈0．05）。PTEN蛋白在食管癌早期表达高于晚期（P〈0．01）。结论：PTEN蛋白在食管鳞状细胞癌组织中的低表达可能与食管鳞癌的发生发展有重要关系。     

食管鳞状细胞癌p16和nm23-H1基因蛋白表达及意义

目的 :探讨多肿瘤抑制基因p16蛋白和nm2 3 H1基因蛋白在人食管癌的表达及与肿瘤生物学行为的关系。方法 :应用免疫组织化学方法观察 51例食管鳞状细胞癌及其淋巴结转移癌中p16蛋白的表达 ,结合观察转移抑制基因nm 2 3 H1的表达。结果 :p16在食管鳞状细胞癌中呈低表达 ( 4 5 1% ,2 3/51) ,阳性率显著低于正常食管粘膜及癌旁组织 (P <0 0 1) ,阳性率与分化程度相关 (P <0 0 1) ,Ⅰ级66 7% ( 14/ 2 1) ,Ⅱ级 34 8% ( 8/ 2 5) ,Ⅲ级 14 3% ( 1/ 7) ;与肿瘤浸润、转移无关。nm 2 3 H1的阳性率为60 8% ( 31/ 51) ,与浸润深度呈负相关 (P <0 0 5) ,淋巴结转移癌的阳性率 ( 18 8% ,3/ 16)显著低于食管癌原发灶 (P <0 0 1)。p16与nm 2 3 H1的表达无关。结论 :p16蛋白在食管癌的低表达提示有频发性的p16基因失活 ,且与肿瘤分化有关。nm2 3 H1与食管癌的浸润 ,转移相关。p16和nm 2 3 H1的失活在食管癌的形成及发展中可能起不同的作用     

检测食管癌标本中PCNA和p16及Ki-67的临床意义

对西京医院进行根治性放疗的食管鳞状细胞癌患者 64例 ,照射剂量DT64～ 70Gy ,进行前瞻性研究 ,对其治疗前活检标本进行SP免疫组化染色 ,计数其阳性细胞数 ,并计算阳性率 ,观察其阳性率与食管癌放疗疗效及预后的关系。结果PCNA阳性率 5 1%～ 67 1% ,中央值 3 2 0 % ,PCNA阳性率 >40 %者较低于此值者预后明显差。p16在低分化癌中较高分化癌阳性表达率低 ,且随着病期的进展阳性表达率降低 ,P <0 0 5 ;Ki 67与近期疗效及预后未见明显差异。初步研究结果提示 ,PCNA阳性率是判断放疗疗效有意义的指标 ,且与p16联合检测对食管鳞癌放疗前恶性程度评定和预后估计有重要意义     

食管鳞状细胞癌中p16、PCNA表达研究

[目的]通过对食管鳞状细胞癌组织中p16与增殖细胞核抗原(PCNA)蛋白表达的研究,探讨二者与食管鳞状细胞癌生物学行为的关系.[方法]检测49例经术后病理证实的食管鳞状细胞癌患者p16与PCNA在不同的病理分级及有无淋巴结转移组中的表达,使用SPSS10.0软件进行统计分析.[结果](1)p16蛋白的阳性细胞面积比率及阳性强度均值在无淋巴结转移组(71.92%,8.38)明显高于淋巴结转移组(60.04%,6.64),P＜0.05.(2)PCNA蛋白的阳性细胞面积比率及阳性强度均值在无淋巴结转移组(42.77%,11.63)明显低于淋巴结转移组(74.67%,15.08),P＜0.01.(3)p16蛋白阳性细胞面积比率及阳性强度均值随病理分级的升高而降低(Ⅰ级79.67%,10.94;Ⅱ级57.50%,9.13;Ⅲ级45.29%,6.889),P＜0.01.(4)PCNA蛋白阳性细胞面积比率及阳性强度均值随病理分级的升高而升高(Ⅰ级38.50%,10.41;Ⅱ级63.63%,13.38;Ⅲ级78.00%,16.13),P＜0.01.[结论]测定p16及PCNA可能对食管鳞状细胞癌的诊断,淋巴结浸润、侵袭及转移,恶性度的判定及治疗有重要作用.     

Circulating endothelial progenitor cells

Angiogenesis research investigates the formation of new blood vessels in wound healing, tumour growth and embryonic development. Circulating, bone marrow-derived endothelial progenitor cells (EPCs) were first described 8 years ago, yet the exact nature of these endothelial precursor cells remains unclear. The contributions of circulating EPCs to angiogenesis in tumours, ischaemic injury and other diseases as well as their usefulness in the repair of wounded hearts and limbs remain under intense investigation.     

慢病毒介导的siRNA沉默Foxp1对肝癌细胞增殖、凋亡和迁移的影响

目的 探讨Foxp1对肝癌细胞增殖、凋亡和迁移的影响。方法 体外合成干扰Foxp1功能的小核酸片段（siRNA Foxp1）,通过重组技术将其插入到慢病毒三质粒系统的转移质粒中,利用包装细胞（293T）将三质粒系统组装为完整的反转录慢病毒载体（lenti-pLL3.7-Foxp1-siRNA）,感染高表达Foxp1的肝癌细胞株。同时构建不含有Foxp1-siRNA序列的直接三质粒系统包装的空病毒载体对照组。荧光显微镜观察慢病毒载体介导siRNA感染细胞的效果。Western blot和Real-time QPCR（RTQPCR）分别检测肝癌细胞中Foxp1蛋白表达和mRNA水平。CCK-8实验、流式细胞仪、细胞划痕愈合实验和侵袭小室实验分别检测细胞增殖、凋亡和迁移的改变。结果 与对照组细胞比较,肝癌细胞感染lenti-pLL3.7-Foxp1-siRNA后,细胞中Foxp1蛋白和mRNA表达显著下降,增殖活性受到显著抑制,细胞凋亡明显增加,在二维空间和三维空间的迁移细胞显著减少（均P<0.01）。结论 Foxp1作为一种多功能转录因子,促进肝癌细胞的增殖和迁移,抑制其凋亡。     

Early diagnosis of oesophageal cancer

Squamous cell carcinoma and adenocarcinoma of the oesophagus are cancers that develop from distinct epithelial sub-types; however, they are both related to chronic inflammation of differing aetiologies. Inflammation leads to somatically inherited genetic mutations altering control of the cell cycle, DNA replication and apoptosis, which together result in autonomous and uncontrolled proliferation. These cancers have often metastasised to lymph nodes and distant organs before symptomatic presentation and therefore carry a poor prognosis. It is therefore vital to diagnose oesophageal cancer at an early stage, before the development of symptoms, when treatment can dramatically improve prognosis. Understanding the pathogenesis of these cancers is vital to guide early diagnostic strategies.     

Detection of Merkel cell virus and correlation with histologic presence of Merkel cell carcinoma in sentinel lymph nodes

Background:

Adjuvant treatment can dramatically improve the survival of patients with metastatic Merkel cell carcinoma (MCC), making early, accurate detection of nodal disease critical. The purpose of this study was to correlate Merkel cell virus (MCV) detection with histopathologic disease in sentinel lymph nodes (SLNs) of MCC.

Methods:

Merkel cell carcinoma cases with SLN (n=25) were compared with negative controls (n=27). Viral load was obtained by quantitative polymerase chain reaction (PCR) for regions VP1 and LT3 of MCV. Histopathologic disease and viral load were correlated.

Results:

Merkel cell virus was detected in 16 out of 17 (94%) of primary MCC (mean viral load (MVL)=1.44 copies per genome). Viral load in the negative controls was <0.01 copies per genome. Merkel cell carcinoma was present in 5 out of 25 (20%) SLN by histopathology, and MCV was detected in 11 out of 25 (44%) MCC SLN (MVL=1.68 copies per genome). In all, 15 out of 25 (60%) SLN showed correlation between histologic and MCV results. In all, 2 out of 25 (8%) samples were histopathologically positive and PCR negative. Of note, 8 out of 25 (32%) samples had detectable MCV without microscopic disease.

Conclusion:

Patients with positive SLN for MCV even if negative by histopathology were identified. The application of molecular techniques to detect subhistologic disease in SLN of MCC patients may identify a subset of patients who would benefit from adjuvant nodal treatment.     

Expression of cyclin D1, D3, E,and p27 in human renal cell carcinoma analysed by tissue microarray

Aberrations in the G1/S transition of the cell cycle have been observed in many malignancies and seem to be critical in the transformation process. Few studies have delineated the presence of G1/S regulatory defects and their clinical relevance in renal cell carcinoma (RCC). Therefore, we have examined the protein contents of cyclin D1, D3, E, and p27 in 218 RCCs, using tissue microarray and immunohistochemistry. The results from a subset of tumours were confirmed by Western blotting and immunohistochemical staining of regular tissue sections. Interestingly, low protein contents of cyclin D1 and p27 were associated with high nuclear grade, large tumour size, and poor prognosis for patients with conventional tumours. We further observed substantial differences in the pattern of G1/S regulatory defects between the different RCC subtypes. The majority of both conventional and papillary cases expressed p27; however, chromophobe tumours generally lacked p27 staining. In addition, conventional RCCs often expressed high cyclin D1 protein levels, while papillary RCCs exhibited high cyclin E. In summary, we have shown that G1/S regulatory defects are present in RCC and are associated with clinico-pathological parameters. The pattern of cell cycle regulatory defects also differed between RCC subtypes.     

间变性大细胞淋巴瘤30例患者外周血内皮祖细胞预后价值分析

目的 探讨间变性大细胞淋巴瘤（ALCL）患者外周血中内皮祖细胞（EPC）的数量变化在疾病预后中的意义.方法 采用流式细胞术检测30例ALCL患者以及对照健康体检者10名外周血中EPC的数量,进行绝对计数,并与临床预后指标国际预后指数（IPI）评分以及间变淋巴瘤细胞激酶（ALK）资料进行对比.结果 ALCL患者治疗前外周血EPC绝对计数为（15.530±28.659）个/μl,与对照组的（0.515±0.294）个/μl相比较,差异有统计学意义（P＜ 0.001）.根据IPI评分标准,把ALCL患者分为低危组（0～1分）、中危组（2～3分）、高危组（4～5分）三组,外周血EPC绝对计数分别为（6.508±7.356）个/μl、（16.830±24.273）个/μl、（21.521±36.057）个/μl,在低危组与中危组、高危组中差异有统计学意义（P＜ 0.01）;中危组与高危组中差异无统计学意义（P＞0.05）;外周血EPC绝对计数在ALK＋与ALK-ALCL患者中分别为（8.367±9.609）个/μl和（22.541±20.845）个/μl,差异有统计学意义（P＜0.01）.ALCL患者外周血EPC绝对计数分为＜20个/μl组与≥20个/μl组,两组间在60周内生存曲线差异有统计学意义.结论 外周血EPC绝对计数与ALCL临床病情进展程度可能有一定的关系,有可能成为评价患者治疗效果及预后的指标之一.     

骨髓成纤维样基质细胞系对急性髓细胞白血病细胞系增殖与分化影响的研究

目的：以急性髓细胞白血病（acute myeloid leukaemia,AML）细胞系为研究对象,观察正常人骨髓成纤维样细胞系（human bone marrow fibro—blastoid stromal cell line,HFCL）对急性单核细胞白血病细胞系U937、急性粒细胞白血病敏感细胞HL-60和多药耐药细胞HL-60／VCR增殖和分化的影响。方法：建立U937、HL-60和HL-60／VCR细胞与HFCL细胞的共培养模型,实验分对照组、直接接触组和transwell组。采用台盼蓝拒染法测定生长曲线;硝基四氮唑蓝（NBT）确定细胞分化;流式细胞仪检测细胞周期和CD11b、CD13、CD14和CD33细胞表面抗原进一步鉴定细胞分化;west-ernblot检测增殖细胞核抗原（PCNA）和P-糖蛋白（P-gP）的表达。结果：与HFCL细胞共培养96h后,U937、HL-60和HL-60／VCR细胞的生长受抑,且与HFCL细胞直接接触组的抑制作用大于用tr-answell组,P〈0．01。同时发现AML细胞系与HFCL细胞共培养后,G0／G1期细胞比例均增高,而S期细胞均减少,P〈0．01;尤其是直接接触组CD11b和CD14表达也均增高（P〈0．01）,CD13和CD33变化不大,NBT阳性细胞轻度增高,且差异有统计学意义。Western blot检测结果显示,3种AML细胞系PCNA表达下调;以直接接触组为甚。提示介导着白血病细胞和骨髓基质细胞间的相互作用的整合素p1（VLA-4）和p2（LFA-1）,在HFCL细胞对AML细胞生长作用影响中起着不可忽视的作用。结论：HFCL对3种具有代表性的AML细胞系U937、HL-60和HL-60／VCR有增殖抑制和诱导分化作用,除了能抑制AML细胞的生长,抑制PCNA的表达,出现G0／G1期阻滞外,还能诱导其部分向单核细胞分化。     

Geldanamycin-mediated inhibition of heat shock protein 90 partially activates dendritic cells,but interferes with their full maturation,accompanied by impaired upregulation of RelB

Background

The chaperon heat shock protein 90 (HSP90) constitutes an important target for anti-tumor therapy due to its essential role in the stabilization of oncogenes. However, HSP90 is ubiquitously active to orchestrate protein turnover, chemotherapeutics that target HSP90 may affect immune cells as a significant side effect. Therefore, we asked for potential effects of pharmacological HSP90 inhibition at a therapeutically relevant concentration on human dendritic cells (DCs) as main inducers of both cellular and humoral immune responses, and on human CD4⁺ T cells as directly activated by DCs and essential to confer B cell help.

Methods

Unstimulated human monocyte-derived DCs (MO-DCs) were treated with the prototypical HSP90 inhibitor geldanamycin (GA). Based on dose titration studies performed to assess cytotoxic effects, GA was applied at a rather low concentration, comparable to serum levels of clinically used HSP90 inhibitors. The immuno-phenotype (surface markers, cytokines), migratory capacity, allo T cell stimulatory and polarizing properties (proliferation, cytokine pattern) of GA-treated MO-DCs were assessed. Moreover, effects of GA on resting and differentially stimulated CD4⁺ T cells in terms of cytotoxicity and proliferation were analysed.

Results

GA induced partial activation of unstimulated MO-DCs. In contrast, when coapplied in the course of MO-DC stimulation, GA prevented the acquisition of a fully mature DC phenotype. Consequently, this MO-DC population exerted lower allo CD4⁺ T cell stimulation and cytokine production. Furthermore, GA exerted no cytotoxic effect on resting T cells, but abrogated proliferation of T cells stimulated by MO-DCs at either state of activation or by stimulatory antibodies.

Conclusion

HSP90 inhibitors at clinically relevant concentrations may modulate adaptive immune responses both on the level of DC activation and T cell proliferation. Surprisingly, unstimulated DCs may be partially activated by that agent. However, due to the potent detrimental effects of HSP90 inhibitors on stimulated CD4⁺ T cells, as an outcome a patients T cell responses might be impaired. Therefore, HSP90 inhibitors most probably are not suitable for treatment in combination with immunotherapeutic approaches aimed to induce DC/T cell activation.     

Characterization of the spectrum of trivalent VAV1‐mutation‐driven tumours using a gene‐edited mouse model

Mutations in the VAV1 guanine nucleotide exchange factor 1 have been recently found in peripheral T cell lymphoma and nonsmall‐cell lung cancer (NSCLC). To understand their pathogenic potential, we generated a gene‐edited mouse model that expresses a VAV1 mutant protein that recapitulates the signalling alterations present in the VAV1 mutant subclass most frequently found in tumours. We could not detect any overt tumourigenic process in those mice. However, the concurrent elimination of the Trp53 tumour suppressor gene in them drives T cell lymphomagenesis. This process represents an exacerbation of the normal functions that wild‐type VAV1 plays in follicular helper T cells. We also found that, in combination with the Kras oncogene, the VAV1 mutant version favours progression of NSCLC. These data indicate that VAV1 mutations play critical, although highly cell‐type‐specific, roles in tumourigenesis. They also indicate that such functions are contingent on the mutational landscape of the tumours involved.