胃癌中Survivin、Cox-2与HIF-1α表达的相互关系及临床意义

目的:探讨胃癌组织中Survivin、Cox-2与HIF-1α蛋白的表达及其临床意义。方法:采用免疫组织化学染色法检测Survivin、Cox-2及HIF-1α在65例胃癌组织、18例癌旁组织及20例正常胃黏膜的表达水平,并分析其与胃癌生物学行为的关系。结果:Survivin、Cox-2与HIF-1α在正常胃黏膜、癌旁组织、癌组织中的表达呈升高趋势,其差异均有统计学意义(P0.05)。Survivin阳性表达在TNM分期、组织学分化程度、胃癌浸润深度、淋巴结转移及远隔转移方面差异显著(P0.05);Cox-2阳性表达在TNM分期、组织学分化程度和淋巴结转移方面差异显著(P0.05);HIF-1α阳性表达在TNM分期、组织学分化程度、胃癌浸润深度和淋巴结转移方面差异显著(P0.05)。相关分析显示,65例胃癌组织中,Survivin、Cox-2与HIF-1α表达均呈正相关(r=0.285,0.405,0.546,P0.05)。结论:Survivin、Cox-2与HIF-1α的过表达在胃癌的发生、发展、浸润和转移过程中发挥协同作用,可成为胃癌基因诊断和治疗的新靶点。     

TrkB/BDNF在非小细胞肺癌组织中的表达及临床意义

目的:探讨肿瘤标记物TrkB/BDNF在非小细胞肺癌癌组织中的表达及其临床意义。方法:采用免疫组化(SP法)和Realtime-PCR方法检测非小细胞肺癌组织和癌旁组织中TrkB/BDNF的mRNA表达水平,并分析其在相关临床因素间的差异。结果:非小细胞肺癌癌组织TrkB/BDNF表达阳性,且明显高于癌旁组织。TrkB/BDNFmRNA水平在有否脑转移和不同TNM分期之间存在明显差异(P<0.05);而在性别、年龄、吸烟、肿瘤大小、分化程度、病理组织学类型间无显著差异(P>0.05)。结论:TrkB/BDNF在非小细胞肺癌癌组织中高表达,并与其脑转移、TNM分期有关,两者可能在非小细胞肺癌的侵袭及转移(尤其脑转移)中发挥重要作用。     

胰岛素样生长因子-1受体和血管内皮生长因子在胃癌中的表达

目的: 探讨胰岛素样生长因子1受体(IGF-1R)和血管内皮生长因子受体(VEGF)的表达与胃癌浸润、转移的关系.方法: 选取胃癌患者56例, 以20例癌旁不典型增生组织及28例正常胃黏膜组织作为对照, 应用SABC免疫组化方法检测IGF-1R和VEGF的表达.结果: IGF-1R蛋白在胃癌组织、癌旁不典型增生组织及正常胃黏膜组织中的阳性表达率分别为87.5%、 55.00%和17.86%, 两两比较, 均有统计学差异(P<0.01);IGF-1R蛋白在胃癌转移组和无转移组中的阳性表达率分别为97.06%和72.73%(P<0.05);浸润至黏膜下层、肌层、外膜的胃癌组织中IGF-1R蛋白阳性表达率分别为66.67%、 92.59%、 100%(P<0.05).VEGF蛋白在胃癌组织、癌旁不典型增生组织及正常胃黏膜组织中的阳性表达率分别为69.64%、 45.00%和25.00%, 两两比较, 均有统计学差异(P<0.05);VEGF蛋白在胃癌转移组和无转移组中的阳性表达率分别为85.29%和45.45%(P<0.01);浸润至黏膜下层、肌层、外膜的胃癌组织中VEGF蛋白阳性表达率分别为40.00%、 74.07%、 85.71%(P<0.05).结论: IGF-1R和VEGF与胃癌的发生、浸润、转移有关, 可联合运用, 作为新的胃癌标记物, 对判断预后有一定价值.     

胃窦癌组织中MMP-26 mRNA及蛋白的表达

目的探讨胃窦癌及癌旁正常胃黏膜组织中MMP-26 mRNA及蛋白的表达意义。方法应用RT-PCR检测40对胃窦癌及癌旁正常胃黏膜组织中MMP-26基因的表达,Western blot方法检测其中10对标本的MMP-26蛋白表达,同时采用免疫组化MaxVision法检测102例胃窦癌组织和58例癌旁胃黏膜组织MMP-26蛋白的表达,分析MMP-26的表达与临床病理变量之间的关系。结果 MMP-26 mRNA在27例胃窦癌中的表达明显高于正常胃黏膜(67.5%)。Western blot结果显示7例胃癌组织中MMP-26蛋白表达明显高于对应的正常胃黏膜组织。免疫组化染色结果显示102例胃癌组织中有57例(55.9%)对MMP-26呈阳性反应,癌旁胃黏膜组织58例中有8例阳性(13.8%)。且MMP-26蛋白表达与胃癌浸润深度、淋巴结转移及TNM分期呈正相关。结论 MMP-26蛋白表达与胃癌浸润转移密切相关,可能作为胃癌早期诊断及预测预后的分子标记物。     

结直肠癌中Survivin和P-gp蛋白表达及其与血清CEA的相关性

目的探讨结直肠癌组织中Survivin和P-gp蛋白表达及其与血清CEA的相关性。方法采用免疫组化SP法检测63例结直肠癌及癌旁黏膜,10例腺瘤组织中Survivin和P-gp蛋白表达,分析两者表达与结直肠癌临床病理特征以及血清CEA值的关系。结果 Survivin和P-gp蛋白在结直肠癌组织中阳性率分别为74.6%、76.19%,均显著高于癌旁和腺瘤组织(P 0.01)。Survivin和P-gp蛋白表达与结直肠癌淋巴结转移、远处转移、TNM分期呈正相关(P 0.05),两者表达之间具有正相关性(P 0.05),Survivin蛋白高表达组血清CEA值明显高于低表达组(P 0.05)。结论 Survivin和P-gp蛋白在结直肠癌中高表达与肿瘤细胞的侵袭、转移密切相关,两者的表达具有协同作用,联合血清CEA检测有助于对肿瘤监测,为治疗结直肠癌提供分子生物学支持。     

胃癌组织中Apollon和Caspase 9的表达及相互关系

目的探讨凋亡相关基因Apollon和Caspase 9在胃癌组织中的表达、临床意义及相关性。方法应用免疫组化SP法检测105例胃癌组织、38例癌旁组织及29例正常胃组织中Apollon和Caspase 9蛋白的表达,并分析Apollon和Caspase 9的表达与胃癌临床病理特征的关系。结果 Apollon在胃癌组织、癌旁组织及正常胃组织中分别有82例(78. 10%)、8例(21. 05%)和2例(6. 90%)阳性,差异有显著性(P 0. 01); Apollon蛋白在胃癌中的表达与肿瘤分化程度、TNM分期及淋巴结转移相关(P 0. 05),与其它临床病理特征无关(P 0. 05); Caspase 9在胃癌组织、癌旁组织及正常胃组织中分别有21例(20. 00%)、23例(60. 53%)和21例(72. 41%)阳性,差异有显著性(P 0. 01); Caspase 9蛋白在胃癌中的表达与肿瘤分化程度、TNM分期及淋巴结转移相关(P 0. 05),与其它临床病理特征无关(P 0. 05); Apollon和Caspase 9在胃癌组织中的表达呈显著负相关(r=-0. 541 1,P 0. 01)。结论 Apollon和Caspase 9的相互作用可能参与胃癌的发生、发展; Apollon与胃癌的浸润、转移密切相关,可作为潜在的治疗靶点。     

胃癌组织中CKS2蛋白表达与患者预后的相关性

目的探讨胃癌组织中CKS2蛋白表达与临床病理特征及患者预后的相关性。方法应用qRT-PCR和免疫组化法检测CKS2基因及蛋白在胃癌及癌旁组织中的表达水平,并探讨其与胃癌患者预后的相关性。结果胃癌组织中CKS2基因的表达水平(0.97±0.16)高于癌旁组织(0.38±0.11,P0.05);胃癌组织中CKS2蛋白的表达水平(21/203,59.60%)与Lauren分型、肿瘤浸润深度、淋巴结转移以及TNM分期密切相关(P0.05)。胃癌患者预后及生物信息学分析结果进一步证实,CKS2蛋白表达水平与胃癌患者不良预后相关(P0.05)。结论 CKS2蛋白在胃癌组织中的高表达水平与胃癌发生、发展密切相关,可作为评估胃癌患者预后的独立因素之一。     

胃癌组织中BMAL1、GLI1的表达及意义

目的探讨BMAL1、GLI1蛋白在胃癌组织中的表达及临床意义。方法采用免疫组化EliVision两步法检测BMAL1、GLI1蛋白在胃癌及癌旁组织中的表达,分析两者表达与胃癌临床病理特征的关系及两者表达的相关性。结果与癌旁组织相比,BMAL1和GLI1蛋白在胃癌组织中的表达上调(P<0.01),阳性率分别为86.7%和83.3%。BMAL1过表达与胃癌淋巴结转移和TNM分期相关(P<0.05),GLI1过表达与胃癌浸润深度、淋巴结转移和TNM分期相关(P<0.05),两者在胃癌中的表达与其余临床病理特征均无关(P>0.05)。BMAL1和GLI1在胃癌中的表达呈正相关(r s=0.526,P<0.001)。结论BMAL1、GLI1与胃癌的发生、发展密切相关,在胃癌演进过程中两者可能起协同作用。     

胃癌中RNA甲基化酶NSUN2的表达及其临床意义

目的探讨胃癌中RNA甲基化酶NSUN2的表达及其临床意义。方法收集109例胃癌采用免疫组化SP法检测癌组织及对应癌旁组织中NSUN2的表达,并分析NSUN2表达与临床病理特征及预后生存时间的关系。利用qRT-PCR及Western blot法检测20对新鲜冷冻胃癌组织及对应癌旁组织中NSUN2的表达。结果 NSUN2在胃癌组织的高表达率为65.1%(71/109),在癌旁组织中的高表达率为15.6%(17/109),两组差异有统计学意义(P0.05)。qRT-PCR和Western blot证实NSUN2 mRNA在胃癌组织中高表达。胃癌中NUSN2高表达与淋巴结转移、浸润深度、远端转移、脉管浸润、神经侵犯、临床分级、TNM分期及患者预后生存时间明显相关(P0.05);与患者年龄、性别、肿瘤位置和直径无关(P0.05)。结论 NSUN2表达与胃癌TNM分期和侵袭性显著相关,NSUN2表达影响患者预后,NSUN2对于胃癌诊断和预后的评判具有一定的价值。     

胃癌中E-cadherin、MMP-2的表达及侵袭转移机制

目的 通过检测E-cadherin和MMP-2蛋白在胃癌中的表达,探讨它们在肿瘤转移和侵袭过程中的作用.方法 取胃癌组织50例,正常胃黏膜10例;应用SP法免疫组化检测E-cadherin、MMP-2蛋白的表达.结果 (1)胃癌组织中E-cadherin蛋白的阳性表达率为46%(23/50),明显低于正常黏膜100%(10/10,P<0.001).转移组阳性表达率为35.29%(12/34)明显低于非转移组68.75%(11/16),差异有显著性(P<0.05).与临床分期、浸润分级的关系显示,Ⅲ-Ⅳ期和T3-T4级癌组织中E-cadherin蛋白表达分别明显低于Ⅰ-Ⅱ期和T1-T2级(P<0.05).E-cadherin表达还与组织学分级相关:胃癌分化程度低则阳性表达率也低;分化程度高则阳性表达率也高,差异有显著性(P<0.05).(2)MMP-2在胃癌组织中阳性表达率为74%(37/50),而正常胃黏膜上皮细胞中几乎无棕染颗粒,其差异有显著性(P<0.05).转移组MMP-2阳性表达率(82.35%)明显高于非转移组(56.25%,P<0.05).MMP-2的表达与组织学分级明显相关:随着胃癌分化程度降低,MMP-2阳性表达率升高(P<0.05);Ⅲ-Ⅳ期胃癌表达明显高于Ⅰ-Ⅱ期(P<0.05);T3-T4级MMP-2阳性表达率高于T1-T2级(P<0.001).(3)MMP-2与E-cadherin表达之间呈高度负相关(相关系数r=-0.368,P<0.009).结论 MMP-2在胃癌中均呈高表达,而E-cadherin则反之.E-cadherin蛋白表达减弱与胃癌侵袭转移潜能相关,可作为胃癌侵袭转移的生物学标志之一.     

The significance of tyrosine kinase receptor B and brain-derived neurotrophic factor expression in salivary duct carcinoma

Salivary duct carcinoma (SDC) is a high-grade salivary gland neoplasm. It may occur de novo or secondarily from pleomorphic adenoma (ex-PA), with secondary development accounting for more than 50% of the cases. In recent years, the expression of tyrosine kinase receptor B (TrkB), which is in the same family as HER2, has been confirmed in various types of carcinomas. However, there are a few studies on SDC. In order to examine the expression and role of TrkB in SDC, we investigated it. Immunohistochemistry was used to detect the expression of TrkB and its ligands, brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4) in 20 patients with SDC. The mRNA levels of TrkB, BDNF, and NT-4 were analyzed using quantitative polymerase chain reaction. TrkB was negative in 10 cases and positive in 10 cases, BDNF was negative in 11 cases and positive in 9 cases, and NT-4 was positive in all cases. There was a high number of TrkB-positive cases in the pT4 group and The H-score of TrkB was also significantly higher in the stage III and IV groups. There was a high number of BDNF-positive cases in the ex-PA group and Histo-score of BDNF had a trend of high expression in ex-PA. There were no significant differences or correlations in mRNA expression. Our results suggest that TrkB may be involved in SDC tumor growth.     

Survivin蛋白表达与胃癌发生发展的关系及意义--组织芯片技术研究

目的探讨凋亡抑制基因Survivin编码蛋白表达与胃癌发生发展的关系及其相关分子病理学机制。方法采用组织芯片仪(M icroarrayer,Beecher Instrum ents,USA)构建含98例胃癌及其癌前病变的直径1.0 mm的225个微组织阵列芯片,免疫组织化学方法检测分析survivin蛋白表达情况。结果胃癌及其癌前病变组织芯片蜡块组织阵列排列整齐,常规切片组织形态良好。凋亡抑制基因survivin编码蛋白在胃癌组织中的表达阳性率显著高于正常胃黏膜、肠上皮化生、不典型增生组织中的表达(P<0.01),胃癌转移组survivin蛋白表达阳性率显著高于无转移组(P<0.05)。Survivin蛋白表达与胃癌病理分期、组织学类型和大体类型没有相关性(P>0.05)。结论组织芯片技术在人胃癌早期诊断中具有其他组织病理学技术不可替代的高效、省时、低消耗的优点。Survivin在胃黏膜癌变过程中被激活而参与胃癌的发生和发展,可作为一个较为理想的肿瘤标志物用于胃癌的早期诊断和转移的预警。     

Changes in survivin messenger RNA level during cisplatin treatment in gastric cancer.

Survivin is a member of the inhibitor of apoptosis protein (IAP) family. The expression of survivin has not been reported in differentiated normal tissues, but it has been observed in many cancerous tissues. Recent studies have revealed that survivin may correlate with the chemo-radio resistance of certain malignant cells. In the present study, the correlation between the occurrence of apoptosis and the level of expression of survivin messenger RNA (mRNA) was investigated in a gastric cancer cell line (MKN-45) and in patients with advanced gastric cancer during cisplatin (CDDP) treatment. In the gastric cancer cell line, the percentage of apoptotic cells (apoptotic index: AI) did not change after 48 h incubation with low-dose CDDP (1 microg/ml), whereas the AI explosively increased between 12 and 24 h treatment with high-dose CDDP (10 microg/ml). Relative levels of expression of survivin mRNA and survivin protein increased after low- and high-dose CDDP treatment. Survivin mRNA was not detected in normal gastric mucosas. Also, in 13 patients with advanced gastric cancer who underwent CDDP-based preoperative chemotherapy, survivin mRNA was detected in only 2 cases (15.4%). Survivin mRNA was observed in the resected tumor specimens of two cases. No significant correlation between survivin mRNA expression and the occurrence of apoptosis in resected tumors or between survivin mRNA expression and patient survival was observed. These findings indicate that survivin may play an important role for the chemoresistance of this cancer cell line. However, the clinical importance of survivin expression remains unclear in patients with gastric cancer.     

Expression and cell localization of brain-derived neurotrophic factor and TrkB during zebrafish retinal development

Brain‐derived neurotrophic factor (BDNF) signaling through TrkB regulates different aspects of neuronal development, including survival, axonal and dendritic growth, and synapse formation. Despite recent advances in our understanding of the functional significance of BDNF and TrkB in the retina, the cell types in the retina that express BDNF and TrkB, and the variations in their levels of expression during development, remain poorly defined. The goal of the present study is to determine the age‐dependent changes in the levels of expression and localization of BDNF and TrkB in the zebrafish retina. Zebrafish retinas from 10 days post‐fertilization (dpf) to 180 dpf were used to perform PCR, Western blot and immunohistochemistry. Both BDNF and TrkB mRNAs, and BDNF and full‐length TrkB proteins were detected at all ages sampled. The localization of these proteins in the retina was very similar at all time points studied. BDNF immunoreactivity was found in the outer nuclear layer, the outer plexiform layer and the inner plexiform layer, whereas TrkB immunoreactivity was observed in the inner plexiform layer and, to a lesser extent, in the ganglion cell layer. These results demonstrate that the pattern of expression of BDNF and TrkB in the retina of zebrafish remains unchanged during postembryonic development and adult life. Because TrkB expression in retina did not change with age, cells expressing TrkB may potentially be able to respond during the entire lifespan of zebrafish to BDNF either exogenously administered or endogenously produced, acting through paracrine mechanisms.     

Immunohistochemical expression of nuclear and cytoplasmic survivin in gastrointestinal carcinoma

Survivin is a protein that is highly expressed in many embryonic tissues, as well as most human tumors. Prior studies have reported both positive and negative correlations between survivin expression and cancer prognosis, but these associations remain controversial. In the present study, we assessed the expression of nuclear and cytoplasmic survivin in gastrointestinal carcinomas. Using these data, we determined the correlation between nuclear and cytoplasmic survivin and, further, investigated correlations between survivin expression and clinicopathological parameters. Seventy-two advanced gastric adenocarcinomas and 78 colorectal adenocarcinomas were analyzed for survivin expression by immunohistochemistry. Expression of both nuclear and cytoplasmic survivin was significantly higher in colorectal carcinomas than in gastric carcinomas (P < 0.01). There was a positive correlation between nuclear and cytoplasmic expression of survivin (r = 0.42, P < 0.001). In gastric carcinomas, the level of survivin protein expression was associated with tumor differentiation, patient age, and lymphatic invasion (P < 0.05, 0.01, and 0.01, respectively). In colorectal carcinomas, the level of nuclear survivin expression was significantly higher in females than in males (P < 0.05). There were no significant associations between survivin expression and most of the clinicopathological parameters. Nevertheless, there was a trend towards an inverse correlation between nuclear survivin expression and tumor aggressiveness in gastric carcinoma; there was a similar trend for cytoplasmic survivin expression. In summary, our results suggest that levels of nuclear and cytoplasmic survivin expression differ between gastric carcinoma and colorectal carcinoma.     

Neurotrophins and neurotrophin receptors in human lung cancer.

The expression of neurotrophins (NTs) and related high- and low-affinity receptors was studied in surgical samples of histologically diagnosed human tumors of the lower respiratory tract. The experiment was conducted with 30 non-small cell lung cancer specimens and in eight small cell lung cancer specimens by Western blot analysis and immunohistochemistry to assess expression and distribution of NT and NT receptor proteins in tissues examined. Immunoblots of homogenates from human tumors displayed binding of anti-nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and NT-3 antibodies as well as of anti-tyrosine-specific protein kinase (Trk) A, TrkB, and TrkC receptor antibodies, with similar migration characteristics than those displayed by human beta-NGF and proteins from rat brain. A specific immunoreactivity for NTs and NT receptors was demonstrated in vessel walls, stromal fibroblasts, immune cells, and sometimes within neoplastic cell bodies. Approximately 33% of bronchioloalveolar carcinomas exhibited a strong membrane NGF and TrkA immunoreactivity, whereas 46% adenocarcinomas expressed an intense TrkA immunoreactivity but a weak immunostaining for NGF within tumor cells. Moreover, squamous cell carcinomas developed an intense TrkA immunoreactivity only within stroma surrounding neoplastic cells. A faint BDNF and TrkB immunoreactivity was documented in adenocarcinomas, squamous cell carcinomas, and small cell lung cancers. NT-3 and its corresponding TrkC receptor were found in a small number of squamous cell carcinomas within large-size tumor cells. No expression of low-affinity p75 receptor protein was found in tumor cells. The detection of NTs and NT receptor proteins in tumors of the lower respiratory tract suggests that NTs may be involved in controlling growth and differentiation of human lung cancer and/or influencing tumor behavior.     

蛇菰多糖降低实验性肝损伤大鼠肝脏BDNF和TrkB的表达

目的蛇菰多糖(BPS)对D-半乳糖(D-gal)所致实验性肝损伤大鼠的肝功能及肝组织内BDNF、TrkB蛋白和凋亡相关蛋白表达的影响。方法将大鼠随机分为对照组(control),D-gal组(皮下注射D-半乳糖200 mg/kg),BPS低(D-gal+BPS-L)、中(D-gal+BPS-M)、高(D-gal+BPS-H)剂量组,分别每天灌胃50、100和200 mg/kg BPS,共6周。心脏取血检测血清谷丙转氨酶(ALT)和谷草转氨酶(AST)水平;HE染色法观察肝组织形态;免疫组织化学染色检测BDNF和TrkB的定位;Western blot检测BDNF、TrkB、Bcl-2、caspase-3和Bax蛋白表达;ELISA测定肝组织内丙醛(MDA)含量和超氧化物歧化酶(SOD)活性。结果与对照组相比,D-gal组肝细胞水肿、点状坏死;血清ALT和AST水平升高(P<0.05),SOD活性降低、MDA含量升高(P<0.05);Bax、caspase-3、BDNF、TrkB表达增加(P<0.05),Bcl-2表达降低(P<0.05);与D-gal组相比,BP...