雌激素受体与食管癌临床病理特性的关系

探讨雌激素受体表达与食管鳞状细胞癌临床病理的关系。方法 用免疫组织化学方法检测了４７例林县食管癌组织中ＥＲ的表达。结果 ４７例食管癌标本中,ＥＲ阳性者１９例,其表达率为４０．４％;生存期大于５年组与小于５年组相比,ＥＲ阳性率为２８．６％和５７．９％,具有显著性差异,与性别,癌组织分化程度及有无淋巴结转移无关。     

Implications of Sex Hormone Receptor Gene Expression in the Predominance of Hepatocellular Carcinoma in Males: Role of Natural Products

The present study was planned to investigate the role of sex hormone receptor gene expression in thepathogenesis of hepatocellular carcinoma (HCC). Adult male Wistar rats were divided into seven groups. Group(1) was negative control. Groups (2), (5), (6), and (7) were orally administered with N-nitrosodiethylamine forthe induction of HCC, then group (2) was left untreated, group (5) was orally treated with curcumin, group(6) was orally treated with carvacrol, and group (7) was intraperitoneally injected with doxorubicin, whereasgroups (3) and (4) were orally administered only curcumin and carvacrol, respectively. The HCC group showedsignificant upregulation in the androgen receptor (AR) and the estrogen receptor-alpha (ERα) gene expressionlevels in the liver tissue. On the contrary, HCC groups treated with either curcumin or carvacrol showedsignificant downregulation in AR and ERα gene expression levels in the liver tissue. In conclusion, the obtaineddata highlight that both AR and ERα but not estrogen receptor-beta (ERβ) gene expression may contribute tothe male prevalence of HCC induced in male rats. Interestingly, both curcumin and carvacrol were found tohave a promising potency in alleviating the male predominating HCC.     

真核表达survivin基因对曲古菌素A诱导卵巢癌细胞凋亡作用的影响

目的　探讨survivin 基因对曲古菌素A( TSA) 诱导卵巢癌细胞凋亡作用的影响。方法　(1) 将本实验室已构建成功的survivin 正义全长真核表达质粒(pcDNA 3. 1-urvivin) ,经脂质体包裹转染人卵巢癌细胞A2780 ,以转染pcDNA 3. 1空载体的A2780 细胞为对照。(2) RT2PCR 和Western blot 方法分别检测survivin mRNA 和蛋白质的表达。(3) MTT 比色法和流式细胞仪(FACS) 分别检测TSA 对两组细胞存活率和凋亡率的影响。(4) Western blot 检测TSA 作用下A2780 细胞中survivin 蛋白的表达变化。结果　(1) RT-PCR和Western blot 检测提示转染pcDNA 3. 1-urvivin 组中survivin mRNA 和蛋白质表达明显高于空载体组。(2) MTT 比色法和FACS 检测提示转染pcDNA 3. 1-urvivin 组细胞存活率明显高于空载体组,细胞凋亡率明显低于空载体组,差异有统计学意义( P<0. 05) 。pcDNA 3. 1-urvivin转染后的2780 细胞对TSA 的敏感性明显降低。(3) Western blot 检测提示survivin 的表达水平随着TSA 作用时间的延长而下降。结论　曲古菌素A 诱导卵巢癌细胞的凋亡作用可能与survivin 基因表达有关。     

RASSF1A基因对肝癌细胞化疗药物敏感性的影响

目的探讨RASSF1A基因的表达对肝细胞肝癌化疗药物敏感性的影响。方法利用已构建成功的稳定表达野生型和突变型RASSF1A基因的肝癌细胞株QGY-7703,化疗药物Mitomycin、Adriamycin、Etoposide、5-Fluorouracilur、Cisplatin分别作用各细胞株后,比较生长抑制率、细胞周期及P53、P21、Bax、Caspase-3蛋白的表达水平。结果野生型RASSF1A的表达可提高Mitomycin诱导的肝癌细胞生长抑制率、凋亡发生率（P<0.05）和Caspase-3的活性,对P53、P21、Bax基因的蛋白表达水平没有影响。结论野生型RASSF1A基因可提高肝癌细胞对Mitomycin的敏感性。     

High Expression of CXCR7 Combined with Alpha Fetoprotein in Hepatocellular Carcinoma Correlates with Extra-hepatic Metastasis to Lung after Hepatectomy

The lung is the most frequent metastatic site of hepatocellular carcinoma (HCC), negatively impactingon survival rates. In this study, we evaluated the prognostic role of the chemokine receptor CXCR7 in lungmetastasis of HCC after hepatectomy, using immunohistochemical detection on tissue microarrays of HCCs,with and without lung metastasis. Using three categories based on staining characteristics, patients with highCXCR7 expression demonstrated a shorter time to development of lung metastasis compared with patients withlow CXCR7 expression (log-rank test) with no effet on overall survival. Analysis of tissue adjacent to tumorshowed patients with microvascular invasion to have higher CXCR7. Stratification based on alpha fetoproteinlevel >20 ng/ml also showed high expression of CXCR7 to be a strong independent prognostic factor. Thesefindings suggest that high expression of CXCR7 in HCCs with elevated alpha fetoprotein levels correlates withmetastasis to lung and poor survival after hepatectomy, indicating potential use as a prognostic factor.     

谷胱甘肽S-转移酶A1在肝癌中的异常表达

目的探讨GSTA1基因在肝癌形成中的作用。方法应用RT-PCR方法检测35例AFB1诱发的树鼩肝癌、癌旁和癌前组织,以及35例人肝癌和癌组织中的GSTA1mRNA表达情况;应用免疫组化方法检测以上组织的GSTA1蛋白表达情况。结果树鼩肝癌组织的GSTA1mRNA和蛋白表达水平均低于癌旁及癌前组织;人肝癌组织的GSTA1mRNA表达水平、蛋白阳性表达率和表达综合得分均显著低于癌旁组织(分别为P<0.001、P<0.05和P<0.001)。结论GSTA1可能是肝癌发生发展的重要相关基因之一;在mRNA水平和蛋白质水平动态观察相关基因在肝癌形成过程中的表达变化有助于阐释肝癌发生的分子机制。     

乳腺浸润性导管癌和浸润性小叶癌细胞粘附分子与雌、孕激素受体表达及意义

目的：探讨乳腺浸润性导管癌（invasive ductal carcinoma,IDC)和浸润性小叶癌（invasive lobular carcinomaILC)组织中细胞粘附分子和雌、孕激素受体（estrogen receptor,ER:progesterone receptor,PR)表达的意义。E -α-catenin,--catenin,γ-catenin和ER、PR的表达。结果：E-cadherin在IDC和ILC中表达缺失和明显减少的分别占18.7%和30%,α-catenin,β-catenin,γ-catenin在IDC中表达缺失和明显减少的分别为75%,43.8%和明显的正相关性;γ-catenin在乳腺浸润性癌中的表达缺失和明显减少与淋巴结转移病例之间有显著的关系。ER和PR在IDC中表达有明显的正相关性。结论：除E-cadherin外,α-catenin,β-catenin,γ-catenin在乳腺浸润性导管癌和浸润性小叶癌中表达明显缺失和减少。γ-catenin表达缺失和明显减少可作为乳腺浸润性癌伴有淋巴结转移的一个预后指标。ER、PR与E-cadherin,α-catenin,β-catenin,γ-catenin可能是乳腺浸润性癌两类独立的预后指标。     

Comparison of Serum Survivin and Alpha Fetoprotein in Egyptian Patients with Hepatocellular Carcinoma Associated with Hepatitis C Viral Infection

Background: Survivin is specific antiapoptotic gene product expressed in a variety of human neoplasmswhose overexpression might assist in early diagnosis and as a prognostic marker. Objectives: The aim was to evaluate the plasma levels of survivin and alpha fetoprotein in patients with chronic hepatitis C viral infection (HCV) with and without hepatocellular carcinoma (HCC). Subjects: 70 subjects were divided into: a control group (Group I) ( 20 healthy volunteers ) and two patients groups: Group II, HCV group (20 patients); and Group III, HCC with HCV(30 patients ). Methods: Thorough physical examination, ultrasonography of the abdomen, laboratory investigations (liver profile, anti-HCV antibodies, hepatitis B surface antigen, Alpha fetoprotein (chemiluminometry) and Survivin (ELISA)) were performed. Results: There was a significant increase in survivin level in HCV patients (Group II) when compared to the control group (p=0.039), along with a significant increase in AFP in Groups II and III when compared to Group I (P<0.001 for both). AFP also distinguished between the two HCV groups. The best generated cut off value for AFP was 10.9 ng/ml and for survivin 13.7 pg/ml. Serum survivin diagnostic sensitivity was 53.3%, diagnostic specificity 62.5% and efficiency 58.6%, in contrast to 100%, 92.5% and 95.7%, respectively, for AFP. Conclusions: While survivin showed significant increase in the HCV group, its diagnostic performance was lower and it proved to be less reliable as a tumor marker for HCC than did AFP.     

Effects of miR-152 on Cell Growth Inhibition,Motility Suppression and Apoptosis Induction in Hepatocellular Carcinoma Cells

Background: miR-152 is involved in the genesis and development of several malignancies. However, its role in HCC has not been fully clarified. The aim of this study was to investigate the clinicopathological significance of miR-152 and its effect on the malignant phenotype of HCC cells. Methods: miR-152 expression was detected using real-time quantitative RT-PCR in 89 pairs of HCC formalin-fixed paraffin-embedded and their adjacent tissues. Functionally, in vitro effects and mechanisms of action of miR-152 on proliferation, viability, caspase activity,apoptosis and motility were explored in HepG2, HepB3 and SNU449 cells, as assessed by spectrophotometry, fluorimetry, fluorescence microscopy, wound-healing and Western blotting, respectively. Results: miR-152 expression in HCC was downregulated remarkably compared to that in adjacent hepatic tissues. miR-152 levels in groups of advanced clinical stage, larger tumor size and positive HBV infection, were significantly lower than in other groups. A miR-152 mimic could suppress cell growth, inhibit cell motility and increase caspase activity and apoptosis in HCC cell lines. Furthermore, Western blotting showed that the miR-152 mimic downregulated Wnt-1, DNMT1, ERK1/2, AKT and TNFRS6B signaling. Intriguingly, inverse correlation of TNFRF6B and miR-152 expression was found in HCC and bioinformatics confirmed that TNFRF6B might be a target of miR-152. Conclusions: Underexpression of miR-152 plays a vital role in hepatocarcinogenesis and lack of miR-152 is related to the progression of HCC through deregulation of cell proliferation, motility and apoptosis. miR-152may act as a tumor suppressor miRNA by also targeting TNFRSF6B and is therefore a potential candidate biomarker for HCC diagnosis, prognosis and molecular therapy.     

雌激素对ER阴性的子宫内膜腺癌细胞JEC增殖的作用

 目的 研究不同浓度的雌激素（17-βE₂）对雌激素受体（ER）阴性的子宫内膜腺癌细胞系JEC增殖和细胞周期的影响。 方法 采用细胞计数法、四甲基偶氮唑蓝比色法（MTT）和流式细胞术（FCM）的方法,观察人子宫内膜腺癌细胞系JEC在加入17-βE₂后的增殖活性和细胞周期时相变化;同时用免疫组化及图像分析,检测JEC细胞在加入17-β E₂前后细胞周期调控蛋白cyclin A表达的变化。 结果 （1）1×10^-7和1×10^-6mol/L的雌激素作用JEC细胞4天后, 细胞计数均明显高于对照组,而其余浓度E₂无明显作用。不同浓度E₂ 作用JEC 24h、48h后,MTT法测OD值和对照组相比无明显差异;1×10^-6mol/L的E₂作用JEC 72h后,其OD值较对照组显著增高;（2）17-β E_2（1×10^-6mol/L）作用JEC细胞72h后使S期及G₂/M期的细胞比例增加,G₀/G₁期细胞比例减少。但作用48h后细胞周期时相分布无明显变化;（3）17-β E₂作用JEC后可使细胞内cyclin A蛋白表达明显增加。 结论 一定浓度的雌激素能促进JEC细胞体外增殖,且具有剂量和时间效应性。研究提示,这种调控作用可能与cyclin A表达的变化有关。     

Effect of Trichostatin A on CNE2 Nasopharyngeal Carcinoma Cells - Genome-wide DNA Methylation Alteration

Trichostatin A (TSA) is a histone deacetylase (HDAC) inhibitor. We here investigated its effects on proliferationand apoptosis of the CNE2 carcinoma cell line, and attempted to establish genome-wide DNA methylationalteration due to differentially histone acetylation status. After cells were treated by TSA, the inhibitory rate ofcell proliferation was examined with a CCK8 kit, and cell apoptosis was determined by flow cytometry. Comparedto control, TSA inhibited CNE2 cell growth and induced apoptosis. Furthermore, TSA was found to inducegenome-wide methylation alteration as assessed by genome-wide methylation array. Overall DNA methylationlevel of cells treated with TSA was higher than in controls. Function and pathway analysis revealed that manygenes with methylation alteration were involved in key biological roles, such as apoptosis and cell proliferation.Three genes (DAP3, HSPB1 and CLDN) were independently confirmed by quantitative real-time PCR. Finally,we conclude that TSA inhibits CNE2 cell growth and induces apoptosis in vitro involving genome-wide DNAmethylation alteration, so that it has promising application prospects in treatment of NPC in vivo. Although manyunreported hypermethylated/hypomethylated genes should be further analyzed and validated, the pointers tonew biomarkers and therapeutic strategies in the treatment of NPC should be stressed.     

转4-1BBL 基因的小鼠肝癌细胞瘤苗刺激脾细胞产生细胞因子的研究

 目的 研究转4-1BBL基因的小鼠肝癌细胞瘤苗体外刺激同系小鼠脾细胞产生细胞因子（IL-2、TNF-α和GM-CSF）的能力。方法 以丝裂霉素C（MMC）处理高表达转m4-1 BBL基因的小鼠Hepa1-6肝癌细胞，制成肿瘤细胞瘤苗（TCV），体外与同系小鼠脾淋巴细胞共同培养后，观察其对脾细胞产生细胞因子（IL-2、TNF-α和GM—CSF）的影响。结果 TCV-4-1 BBL刺激后，脾细胞体外分泌细胞因子IL-2、TNF-α和GM-CSF的水平明显增高。结论 转4-1BBL基因的小鼠肝癌细胞瘤苗能刺激脾细胞产生细胞因子IL-2、TNF-α和GM-CSF。     

siRNA干扰MTH1基因对肝癌HepG2细胞凋亡和迁移的影响

目的 应用siRNA干扰技术抑制人肝细胞癌系HepG2细胞MTH1基因表达,并研究MTH1基因干扰后对HepG2细胞凋亡和迁移能力的影响。方法 设计并化学合成3对特异性MTH1-siRNA,采用阳离子脂质体法瞬间转染肝癌HepG2细胞,离体培养肝癌HepG2细胞,并设正常对照组、阴性对照组及MTH1-siRNA1、MTH1-siRNA2、MTH1-siRNA3转染组。采用蛋白印迹（Western blot）法检测转染MTH1-siRNA后HepG2细胞MTH1蛋白水平表达的变化,并筛选干扰效果最佳序列。运用流式细胞技术和划痕实验检测转染siRNA-MTH1后HepG2细胞凋亡、体外迁移能力的变化。结果 MTH1-siRNA3转染组干扰效果与正常组和阴性对照组相比效果具有统计学意义（P=0.002, P=0.005）。MTH1-siRNA3转染组细胞凋亡率高于正常组及阴性对照组,差异有统计学意义（P=0.012, P=0.013）;MTH1-siRNA3转染组细胞的划痕24、36 h愈合率均低于正常组和阴性对照组,差异有统计学意义（P=0.001, P=0.000）。结论 MTH1在肝癌细胞的凋亡及迁移中起重要作用,干扰MTH1的表达能促进肝癌细胞凋亡并降低肝癌细胞的迁移能力。     

ER、PR及PCNA在子宫内膜增生过长和内膜腺癌中的表达

为了探讨女性激素在子宫内膜腺癌和子宫内膜增生过长发病中的作用，为临床内分泌治疗提供理论依据，作者采用免疫组织化学ＳＰ法，对手术切除和诊刮的１７例子宫内膜腺癌和４０例子宫内膜增生过长标本进行ＥＲ、ＰＲ和ＰＣＮＡ含量检测，并结合组织学类型和分化程度进行分析。结果显示内膜腺癌组织中ＥＲ、ＰＲ的含量低于增生期和增生过长内膜组织（Ｐ＜０．０５），与肿瘤的组织学类型和分化有关。ＰＣＮＡ在子宫内膜腺癌中的含量显著高于非肿瘤性内膜组织（Ｐ＜０．０１），与分化程度呈负相关。与ＥＲ、ＰＲ的含量无明显相关性。结果表明ＥＲ在子宫内膜增生过长和内膜腺癌的发病中起重要作用，ＰＣＮＡ的过度表达可能与细胞的异常生长和分化有关。     

转化生长因子α及其受体在人脑星形细胞瘤中的表达及意义

目的　研究转化生长因子α(transforminggrowthfactoralpha ,TGF α)及其表皮生长因子受体 (epidermalgrowthfactorreceptor ,EGFR)在人脑星形细胞瘤中的表达及意义。方法　采用免疫组织化学方法检测 5 0例人脑星形细胞瘤标本和 10例正常人脑组织中TGF α和EGFR蛋白表达。结果　 5 0例人脑星形细胞瘤TGF α和EGFR蛋白表达总阳性率分别为 64 .0 % ( 3 2 /5 0 )和 68.0 % ( 3 4/5 0 ) ,正常脑组织未见阳性表达 (P <0 .0 1) ;二者密切相关 (P <0 .0 1) ,并与星形细胞瘤病理分级有显著相关性 ,其中Ⅰ～Ⅱ级者阳性率显著低于Ⅲ、Ⅳ级者 (P <0 .0 5 ) ;结论　TGF α和EGFR形成的自分泌环在人脑星形细胞瘤发生发展过程中起重要作用 ,同时检测它们可作为判断人脑星形细胞瘤临床分期、预后的重要指标     

2-甲基萘[2,3-b]呋喃-4,9-酮(FNQ3)对肺腺癌细胞株(A549)细胞Fas受体表达和细胞凋亡的影响

目的探讨2-甲基萘[2,3-b]呋喃-4,9-酮(FNQ3)对A549细胞表面Fas受体表达和细胞凋亡的调节效应。方法给A549细胞投与FNQ3,从药物的浓度依赖性及投药的时间依赖性两方面来观察药物对A549细胞表面Fas受体表达及诱导凋亡的作用。用半定量RT-PCR法检测FasmRNA,用流式细胞仪检测A549细胞表面的Fas受体表达和细胞凋亡。结果A549细胞表面Fas受体表达和A549细胞FasmRNA及细胞凋亡敏感度都随着药物浓度和投药时间的增加而增加。在研究药物浓度依赖性时,FNQ3浓度为2.5μg/ml时FasmRNA产量相对较高,为75000分子/μl;A549细胞表面Fas受体表达相对较多;细胞凋亡率达到37.26%(P<0.01)。在研究药物时间依赖性时,FNQ3(0.5μg/ml)投药时间为7天时,FasmRNA产量相对较高,为150000分子/μl;A549细胞表面Fas受体表达相对较多;细胞凋亡率30.45%(P<0.02)。结论FNQ3作为1种抗肿瘤药物能够诱导A549肿瘤细胞表面Fas受体表达,并上调由Fas受体介导的细胞凋亡。     

姜黄素诱导人结肠癌细胞株SW480凋亡及其bcl-2表达相关性研究

目的探讨姜黄素对人结肠癌细胞株SW480诱导凋亡的影响及其作用机制。方法5～50μM姜黄素分别处理SW480细胞6～48h后,MTT法检测细胞的生长活性,流式细胞仪检测细胞周期,TUNEL法检测细胞凋亡,免疫组化法检测细胞内bcl2蛋白表达水平。结果MTT显示姜黄素对SW480细胞具有细胞毒作用,呈时间浓度依赖性。FCM结果显示细胞周期中G0/G1期细胞比例增加,S期、G2/M期比例下降。TUNEL法显示凋亡细胞数增加,免疫组化结果显示bcl2基因表达明显减弱。结论姜黄素能抑制结肠癌SW480细胞的增殖并诱导细胞凋亡。其机制可能与干扰细胞周期及下调bcl2基因表达有关。     

ESM-1作为肝癌血管内皮标志物的意义

目的研究内皮细胞特异性分子1（ESM-1）在肝细胞癌（HCC）中的表达及其意义。 方法利用表达谱芯片技术比较3例HCC和癌旁肝组织基因表达的差异,筛选出与肿瘤内皮细胞功能有关的基因群。采用免疫组织化学SABC法检测100例HCC及其癌旁肝组织和16例正常肝组织中ESM-1的表达。 结果表达谱芯片技术获得112条差异表达基因,其中与内皮细胞功能有关的基因6条（包括ESM-1）。ESM-1特异性表达于HCC的血管内皮细胞,而癌旁肝组织和正常肝组织的血管内皮则几乎不表达。ESM-1的表达水平和HCC的镜下静脉浸润密切相关（P<0.05）。ESM-1高表达组的无瘤存活率低于ESM-1低表达组,差异具有统计学意义（P<0.05）。 结论ESM-1与HCC的恶性生物学行为、血管生成以及不良预后密切相关。