Syk基因在食管鳞癌组织中的表达及临床意义

目的:分析食管鳞癌组织及其对应癌旁组织中Syk蛋白和mRNA的表达,探讨其与食管鳞癌临床恶性生物学行为的关系.方法:采用免疫组织化学法检测48例食管鳞癌组织及其对应癌旁组织中Syk蛋白的表达;RT-PCR法检测143例食管鳞癌组织及其对应癌旁组织中PSyk mRNA的表达,并观察其与食管鳞癌患者肿瘤大小、TNM分期、淋巴结转移的关系.结果:食管鳞癌组织Syk蛋白表达阳性率显著低于癌旁组织(16.67% vs 89.58%,P<0.05);Syk蛋白表达与肿瘤删分期相关(X<'2>=6.713,P<0.05):Syk在淋巴结转移组中的阳性表达率显著低于无淋巴结转移组(3.03% vs 29.41%,P<0.05);Syk表达与肿瘤大小无关(X<'2>=0.017,P>0.05).食管鳞癌组织中Syk mRNA的表达量明显低于其在对应癌旁组织中的表达量(t=-11.27,P<0.05).结论:Syk蛋白和mRNA在食管鳞癌中表达缺失与其发生及转移倾向相关,Syk可能是食管鳞癌的肿瘤抑制基因,可能作为分子标记而用于食管鳞癌的早期诊治.     

STAT3信号蛋白与食管鳞癌上皮间质转化的关系及其意义

目的:探讨食管鳞癌中STAT3、p-STAT3蛋白表达与食管鳞癌上皮间质转化的关系及其在食管鳞癌浸润转移中的作用.方法:采用免疫组织化学技术检测80例食管鳞癌中STAT3、p-STAT3及上皮间质转化标志物E-cadherin和Vimentin的表达.结果:食管鳞癌组织中STAT3、P-STAT3、E-cadherin和Vimentin蛋白阳性率与癌旁正常组织相比有显著性差异(STAT3: 87.5% vs 70.0%; p-STAT3: 72.5% vs 28.8%; E-cadherin:37.5% vs 78.8%; Vimentin: 48.8% vs 0%,均P<0.01).食管鳞癌组织STAT3、p-STAT3的表达均与E-cadhcrin的表达呈负相关(r=-0.41 0,-0.506;均P=0.000),与Vimentin表达均呈正相关(r=0.293,0.321;P=0.008,0.004),且与癌组织的浸润深度密切相关(均P<0.05).结论:信号蛋白STAT3可能参与了食管鳞癌的上皮间质转化过程,并与食管鳞癌的侵袭转移相关.     

lncRNA HOTAIR在食管鳞癌患者血清中的表达及其意义

背景由于缺乏食管早期诊断、转移复发检测和预后判断的高效生物标志物,导致食管鳞癌晚期确诊,生存率低.lncRNA HOTAIR在癌组织中一致高表达,并且与肿瘤的发生发展有关,但其在血清中的研究较少.目的探讨lncRNA HOTAIR在健康志愿者和食管鳞癌血清中的表达差异及其临床意义.方法采用RT-qPCR分别检测48例健康志愿者血清、48例食管鳞癌初治患者血清及其配对癌组织中lncRNA HOTAIR表达水平.秩和检验分析健康志愿者和食管鳞癌患者血清中HOTAIR表达水平及其差异性:受试者工作曲线(receiver operating characteristic,ROC)对血清lncRNA HOTAIR诊断效能进行评价:食管鳞癌患者血清与其配对癌组织中表达水平相关性采用Spearm an分析.结果 lncRNA HOTAIR在食管鳞癌患者血清中表达水平显著高于健康志愿者,P=0.0099,ROC曲线下面积为0.8618,敏感度为0.7612,特异度为0.9091,截断值为14.4670,提示血清中lncRNA HOTAIR表达水平对食管鳞癌的诊断敏感度和特异度均较满意:食管鳞癌患者血清与其配对癌组织中lncRNA HOTAIR的表达水平呈正相关(r_s=0.3920,P=0.0124):其与食管鳞癌患者临床病理因素相关性的研究发现,伴远处转移食管癌患者血清HOTAIR表达水平显著高于无转移者P=0.003:伴随TNM分期渐晚,lncRNA HOTAIR表达水平逐渐升高,具有等级相关性,P=0.011.结论血清lncRNA HOTAIR及其动态变化有望成为食管鳞癌患者早期诊断、判断病情严重程度,预测治疗效果及预后生物学指标.     

miR-143在食管鳞癌和上皮内瘤变组织中的表达及其临床意义

背景:研究发现microRNA在肿瘤的发生、发展过程中起癌基因或抑癌基因的作用。然而,microRNA-143(miR-143)在食管鳞癌中的作用需进一步研究。目的:探讨食管鳞癌组织中miR-143表达及其临床意义。方法:选取2013年1月—2013年12月南京医科大学第一附属医院63例食管鳞癌及其癌旁正常组织以及40例食管上皮内瘤变及其相应正常组织,应用实时定量PCR法检测miR-143表达,并分析其与临床病理特征的相关性。结果:与对照组相比,食管鳞癌和上皮内瘤变组织中miR-143表达均明显下调(P0.05)。miR-143表达与患者的病理类型有关(P0.001),而与性别和年龄无关(P0.05)。miR-143表达与食管鳞癌患者病理分级、淋巴结转移、TNM分期有关(P0.05),与肿瘤浸润深度无关(P0.05)。结论:miR-143在食管鳞癌、食管上皮内瘤变组织中的表达均下调,可能与食管鳞癌发生、发展密切相关,有望成为诊断食管鳞癌的新靶点。     

金属蛋白酶抑制基因RECK在食管鳞癌中的表达及生物学意义

目的: 研究RECK基因在食管鳞癌组织中的表达情况及其与食管鳞癌发生、发展的关系.方法:?2006-02-26/2006-03-16河南省安阳市肿瘤医院食管癌手术切除标本62例, 所有病例术前均无化疗、放疗及免疫治疗史. 全部病理组织学证实均为鳞状细胞癌. 全部样本分别在癌灶、癌旁3 cm以内及远端正常黏膜组织分别取材62、31和62例. 采用免疫组化SP法和原位杂交方法进行RECK蛋白及mRNA的检测.结果: 食管鳞癌组织中RECK蛋白及mRNA表达均与癌的组织学分级、浸润深度及淋巴结转移密切相关; 在食管鳞癌癌变过程中RECK蛋白及mRNA表达在癌组织、癌旁不典型增生组织及正常黏膜组织中的表达率依次增高, 组间比较有明显差异(RECK蛋白: χ2=10.331, P<0.01; RECK mRNA: χ2 = 19.186, P<0.01); RECK蛋白与mRNA的表达呈正相关关系(r = 0.416, P<0.01).结论:RECK低表达与食管鳞癌的发生、发展有关, RECK可作为食管鳞癌早期诊断的辅助指标.     

实时荧光定量PCR技术检测食管鳞癌c-myc mRNA的表达

目的 探讨c-myc mRNA表达与食管鳞癌发病的关系.方法 采用实时荧光定量PCR法检测24例食管鳞癌患者食管鳞癌组织和正常食管鳞状上皮组织、癌旁食管鳞状上皮组织中c-myc mRNA的表达.结果 食管鳞癌组织与癌旁食管鳞状上皮组织c-myc基因的表达量相近,二者均明显高于正常食管鳞状上皮组织(P<0.05).结论 c-myc基因在食管鳞状上皮癌变过程的早期阶段起重要作用.     

c-Met基因在新疆哈萨克族汉族 食管鳞癌的表达及其与临床病理的相关性研究

目的分析c-Met基因在新疆哈萨克族(哈族)、汉族食管鳞癌患者中的表达及其与临床病理的相关性。方法收集2013年5月至2014年5月新疆医科大学附属肿瘤医院收治且手术切除的100例冰冻食管鳞状细胞癌组织以及配对对癌远端正常组织的标本,其中哈族50例,汉族50例。观察两组c-Met基因转录的m RNA的表达相关情况。结果在本次研究完成后我们发现,c-Met基因转录的m RNA在所有患者的食管鳞癌组织中共50例呈现阳性表达,其中汉族40例,哈族60例,配对癌远端的正常组织中6例呈现出阳性表达的情况,其中汉族2例,哈族4例,对于食管鳞癌组织及其配对的癌远端的正常组织相比,c-Met基因的表达呈现出明显增加的情况,并与肿瘤细胞的分化程度有较大的相关性。结论对于食管鳞癌患者而言,c-Met基因在部分的食管鳞癌的组织中会呈现出一种高表达的情况,同时与患者的性别、年龄以及肿瘤的发生部位并无显著的相关性,但与患者的肿瘤分化程度有着明显的相关性,同时c-Met的表达在哈族以及汉族患者的食管鳞癌中的表达差异无统计学意义。     

TMEM16A在食管鳞癌中的表达及意义

目的: 探讨TMEM16A在食管鳞状细胞癌发生发展中的意义.方法: 2007-01/2008-04南京医科大学附属江宁医院手术切除的食管鳞癌标本60例, 均为癌组织及距癌至少5 cm远的配对癌旁组织. 应用免疫组织化学SP法及Western blot法检测食管鳞癌组织及相应癌旁组织的TMEM16A蛋白表达.结果: 免疫组织化学结果: 食管鳞癌组织、癌旁组织中TMEM16A蛋白的表达差异具有统计学意义(52.50% vs 20.00%, P<0.05),TMEM16A在淋巴结转移组中的阳性表达率显著高于无淋巴结转移组(65.38% vs 21.43%,P<0.05), 不同分化程度高分化、中分化、低分化的食管鳞状细胞癌中, TMEM16A的阳性表达率分别为27.27%、43.75%和84.62%,3者比较差异有显著性( P<0.05). TMEM16A表达与食管鳞癌的TNM分期无明显相关性;Western blot法检测发现TMEM16A在食管癌组织的表达较癌旁组织强( t = 42.16, P<0.05).结论: TMEM16A可能在食管癌的发生中有一定的作用, 有望为食管癌的分子诊断、基因治疗和预后评估提供新的分子靶点.     

PTEN在食管癌中的表达及其与微血管密度和临床病理特征的关系

目的:探讨抑癌基因PTEN在食管癌及癌旁正常组织中的表达, 及其与微血管密度(MVD)和临床病理特征的关系.方法:选用食管鳞癌手术切除病理标本48例,手术远端正常食管组织标本40例. 采用免疫组织化学SP法检测PTEN编码蛋白的表达水平,CD31抗体进行血管内皮染色、计算微血管密度, 分析PTEN在不同组织中的表达与MVD和食管癌组织分化程度、浸润深度、淋巴结转移的关系.结果:食管鳞癌组织中PTEN编码蛋白阳性表达率低于癌旁正常食管黏膜组织(52.08%vs 92.50%, P<0.01), 而其MVD值显著高于癌旁正常组织(41.72±8.67 vs 21.01±3.85, P<0.01); Ⅰ、Ⅱ、Ⅲ级鳞癌PTEN阳性表达率有显著差异(75.0% vs 55.0% vs 33.33%, 均P<0.05), MVD值差异无统计学意义; 癌组织侵及浅肌层以上与深肌层PTEN与MVD值的表达有显著差异(77.27% vs 42.31%; 35.49±5.89vs 46.01±6.27, 均P<0.01); 淋巴结转移组与非转移组PTEN阳性表达率无显著差异, MVD值差异则有统计学意义(46.71±7.89 vs 35.92±2.54, P<0.01).结论:抑癌基因PTEN、MVD在食管癌中表达的高低, 与肿瘤的生长、浸润和转移相关.PTEN基因表达的突变或缺失能促进肿瘤血管的形成, 可作为临床治疗和判断预后的依据.     

c-IAP1在食管鳞癌中的表达及其对化疗敏感性的影响

目的:探讨食管鳞癌细胞凋亡抑制蛋白1(c-IAP1)表达与化疗敏感性的相关性.方法:食管鳞癌组织芯片免疫组织化学染色,分析食管鳞癌组织及其配对癌旁食管上皮中c-IAP1的表达和定位及其与肿瘤,临床分级的关系.免疫印迹分析食管癌细胞C-IAP1和Smac表达,用RNA干扰技术敲降Smac表达,MTT法检测细胞对化疗药物敏...     

Correlation between expression of human telomerase subunits and telomerase activity in esophageal squamous cell carcinoma

AIM: To investigate telomerase activity and hTERT, TP-1 expression and their relationships in esophageal squamouscell carcinoma (ESCC).METHODS: Telomerase activity was measured in 60 ESCCtissues using telomeric repeat amplification protocol (TRAP)assay by silver staining. In situ hybridization was used for detecting hTERT and TP-lmRNA.RESULTS: The telomerase activity was detected in 83.3 % of ESCC tissues. The difference of telomerase activity was significant between well and poorly cancer differentiated lesions (P<0.05). The positive rate of telomerase activity was higher in patients with lymphatic metastasis than in patients without lymphatic metastasis. In cancer tissues hTERT mRNA expression was 75 % and TP-1 mRNA expression was 71.7 %. The expression of hTERT, TP-1 mRNA in well and poorly differentiated carcinoma was not significant. The expression of hTERT mRNA was correlated with telomerase activity, but TP-1 mRNA expression was not correlated with it.CONCLUSION: Telomerase activity and hTERT, TP-1 mRNA expression are up-regulated in ESCC. Telomerase activity in ESCC is correlated with lymphatic metastasis and cancer differentiation. Telomerase activity may be used as a prognostic marker in ESCC. hTERT mRNA expression is correlated with telomerase activity. Enhanced hTERT mRNA expression may initially comprehend the telomerase activity level, but it is less sensitive than TRAP assay.     

Expression and clinical significance of S100A2 and p63 in esophageal carcinoma

AIM： TO investigate the expression and clinical significance of S100A2 mRNA and protein, p63 protein in esophageal squamous cell carcinoma （ESCC） and their roles in carcinogenesis and progression of esophageal carcinoma （EC）. METHODS： Immunohistochemical staining （S-P method） for S100A2 and p63 protein were performed in 40 samples of ESCC and 40 samples of normal esophageal mucosa. In situ hybridization （ISH） was used to detect the expression of S100A2 mRNA. RESULTS： Expression of S100A2 mRNA in ESCC was positive in 77.5% of samples, which was lower than that in normal mucosa （100%） by ISH （P = 0.002）. The expression level of S100A2 mRNA was closely related to differentiation and and node-metastasis （P = 0.012, P = 0.008）. Expression of $100A2 protein was positive in 72.5% of ESCC samples and expression of p63 protein was positive in 37.5% of ESCC samples, and was lower than that in normal mucosa （100%） （P = 0.000）. The expression of S100A2 protein was correlated with the differentiation and node-metastasis （P = 0.007, P = 0.001）, but no relationship was observed between the expression of p63 protein and clinical pathological manifestations. S100A2 protein was positively correlated with the expression of S100A2 mRNA, and negatively associated with the expression of p63 protein （P = 0.000, P = 0.002）. CONCLUSION： S100A2 and p63 protein both play important roles in the carcinogenesis of ESCC. An investigation into the combined expression of S100A2 and p63 may be helpful in early diagnosis and in evaluating the prognosis of ESCC.     

Expression of vascular endothelial growth factor-C and angiogenesis in esophageal squamous cell carcinoma

AIM: To investigate the expression of vascular endothelial growth factor-c (VEGF-C) mRNA and microvessel density (MVD) in human esophageal squamous cell carcinoma (ESCC) and its relationship with clinical significance. METHODS: Specimens obtained from 43 patients undergoing surgical resection for ESCC were used in this study. The expression of VEGF-C mRNA was examined by in situ hybridization. Tumor MVD was determined immunohistochemically with anti-CD31 antibody and estimated by image analysis. Ten sections of adjacent normal mucosa were also examined. RESULTS: VEGF-C mRNA expression was detected in cytoplasm of carcinoma cells. Of the 43 ESCC patients studied, 18 cases (41.9%) were positive for VEGF-C mRNA. No VEGF-C mRNA expression was observed in normal esophageal mucosa. VEGF-C mRNA expression correlated significantly with lymph node metastasis, TNM stage and depth of invasion (P < 0.05). Furthermore, histological grade (differentiation) tended to correlate with VEGF-C mRNA expression, but was not statistically significant (P > 0.05). In tumor lesions, the MVD was significantly greater than that in normal esophageal mucosa. MVD correlated significantly with lymph node metastasis, TNM stage and depth of invasion (P < 0.05), but not with histological grade (differentiation) (P > 0.05). Lesions with VEGF-C mRNA expression had a significantly higher MVD than that of those without VEGF-C mRNA expression (P < 0.05). CONCLUSION: VEGF-C plays a role in lymphatic metastasis via lymphangiogenesis and angiogenesis in ESCC. VEGF-C is one of the important predictors of the biological behavior in ESCC.     

Association between Bmi1 and clinicopathological status of esophageal squamous cell carcinoma

AIM： To investigate the clinicopathological roles of Bmil in esophageal squamous cell carcinoma （ESCC）.METHODS： Quantitative real-time polymerase chain reaction and immunohistochemical staining for Broil were performed in cancerous and adjacent non-cancerous paraffin-embedded esophageal specimens.RESULTS： The Bmil expression level was unaffected by gender and age. The level of Broil mRNA in ESCC was significantly higher than that in the adjacent non-cancerous tissues （2.181 ± 2.158 vs 0.931 ± 0.894, P = 0.0152）, and its over-expression was aggressively associated with lymph node metastasis （3.580 ± 2.487 vs 1.703 ± 0.758, P = 0.0003）, poorer cell differentiation （P = 0.0000） and advanced pathological stage （3.827± 2.673 vs 1.590 ± 0.735, P = 0.0001）. The patients were divided into high-expression and low-expression groups based on the median expression level of Bmi1 mRNA, and a shorter overall survival time in the former group was observed. Immunohistochemistry for Bmi1 oncoprotein showed diffusely positive, focally positive and negative expression in 44, 16 and 10 of 70 ESCC cases, respectively, compared with three, two and five of 10 adjacent non-cancerous cases （P = 0.027）. The positive rate of the oncoprotein in samples of histological grade Ⅲ was higher than that of grade Ⅱ（P = 0.031）, but its expression had no relation to the lymph node metastasis and pathological staging. In 70 ESCC samples, Bmi1 showed high intense expression in the cytoplasm and less or even no expression in the nucleus.CONCLUSION： Bmi1 was over-expressed in ESCC. Increased Bmi1 mRNA expression was significantly associated with ESCC progression, and the oncoprotein was largely distributed in the cytoplasm of tumor cells.     

Kiss-1在食管鳞状细胞癌组织中的表达及临床病理意义

目的:观察Kiss-1蛋白及mRNA在食管鳞状细胞癌组织中的表达及其临床病理意义.方法:采用免疫组织化学SP法、原位杂交和逆转录聚合酶链反应(RT-PCR)技术,联合检测了62例食管鳞状细胞癌、31例癌旁不典型增生及62例正常食管组织中Kiss-1蛋白和mRNA的表达,并探讨其临床病理意义.结果:在食管鳞状细胞癌组织、癌旁不典型增生及正常食管组织中,Kiss-1蛋白的阳性表达率分别为56.5%、67.7%、90.3%(P<0.05);用原位杂交检测Kiss-1 mRNA阳性表达率分别为51.6%、74.2%、95.2%(P<0.05);用RT-PCR技术检测Kiss-1 mRNA阳性表达率分别为54.8%、71.0%、88.7%(P<0.05);食管鳞状细胞癌组织中Kiss-1蛋白及mRNA的低表达与淋巴结转移密切相关(P<0.05),而与患者的性别、年龄、肿瘤的组织学分级无关(P>0.05);Kiss-1蛋白的低表达与浸润深度有关(P<0.05);在食管鳞状细胞癌组织中,Kiss-1蛋白及mRNA的表达呈正相关(P<0.05),用原位杂交及RT-PCR技术对Kiss-1 mRNA表达情况的检测结果也呈正相关(P<0.05).结论:Kiss-1基因的表达降低或缺失和食管鳞状细胞癌的发生发展及转移有关,有望成为食管鳞状细胞癌早期诊断和预后判断的重要指标之一.     

Cyclic adenosine 3',5'-monophosphate negatively regulates clusterin gene expression in Leydig tumor cell lines.

The clusterin protein and its messenger RNA were identified in many tissues including testis. In this report, we demonstrate the expression of clusterin gene in four Leydig tumor cell lines, including mouse MA-10 and I-10 and rat R2C and LC-540. When the cells were incubated with 0.1 mM 8-bromo-cAMP or (Bu)2cAMP for 17 h, an unexpected, profound suppression of clusterin mRNA accumulation was observed. A 60-70% decrease in clusterin mRNA was observed in MA-10 and R2C cells, 10% in I-10 cells, and no apparent change in LC-540 cells. The inhibitory effect of cAMP was specific to the clusterin gene, since in the same cells cholesterol side-chain cleavage enzyme mRNA was drastically elevated in MA-10 and I-10 cells while alpha-tubulin mRNA levels were not changed in all four cell lines. The reduction could be detected as early as 4 h, and was evident at 17 h after cAMP administration. Removal of cAMP from culture media at 17 h prevented the decline of clusterin mRNA. The suppression of clusterin gene expression can also be demonstrated by treatment with human CG or forskolin, which were known to elevate intracellular cAMP levels. Our observations suggest: 1) cAMP negatively regulates clusterin gene expression in two Leydig tumor cell lines, MA-10 and R2C; 2) The inhibitory effect of cAMP on clusterin gene expression is probably acting through the protein kinase A pathway; and 3) The four Leydig tumor cell lines respond differently to cAMP in the expression of clusterin and side-chain cleavage genes.     

Expression level of beta-catenin is associated with prognosis of esophageal carcinoma

AIM: To examine the association of beta-catenin with the clinicopathologic features and prognosis of esophageal squamous cell carcinoma (ESCC). METHODS: Beta-catenin mRNA expression level in 40 ESCC patients (28 males and 12 females, age range 38-82 years, median 60 years) was analyzed by real- time PCR. Beta-catenin mRNA expression levels in tumor cells were categorized as weaker (level 1) or equal to or stronger (level 2) than those in endothelial cells. We examined the correlation between the beta-catenin expression and the clinicopathological factors and prognosis of ESCC patients. RESULTS: Level 2 beta-catenin expression was found in 29 patients. ESCC with level 2 expression had a higher rate of lymphnode metastasis (0.0776 ± 0.0369 vs 0.3413 ± 0.1803, P < 0.001) and deeper tumor invasion (0.0751 ± 0.0356 vs 0.3667 ± 0.1928, P < 0.001), and a poorer survival rate (P = 0.0024) than ESCC with level 1 expression. CONCLUSION: Beta-catenin expression in ESCC is of great importance.