胃癌survivin基因mRNA和蛋白的表达与临床病理关系

目的　研究胃癌组织中survivin基因的mRNA和蛋白表达情况及其与临床病理参数的关系。方法　应用原位杂交方法和免疫组化SP法检测 5 4例胃癌 ,34例胃良性病变 ,2 0例胃正常组织标本中survivin基因mRNA及蛋白的表达。并对其与临床病理因素和二者的关系进行分析。结果　Survivin蛋白表达阳性率在胃癌、良性疾病组织和正常组织分别为 87 0 % (47/ 5 4 )、 2 3 5 % (8/ 34)和 15 % (3/ 2 0 ) ;SurvivinmRNA阳性率为 79 6 % (43/ 5 4 )、 2 3 5 % (8/ 34)和2 0 % (4/ 2 0 )。胃癌组远大于正常胃组织和良性疾病组 ,而正常胃组织与胃良性疾病组之间差异无显著性。SurvivinmRNA与蛋白在胃癌中的表达呈正相关 (rs=0 6 79,P <0 0 5 )。且其阳性率高低与性别、年龄、组织学类型无关 ;而与淋巴结转移、TNM分期、组织学分级有关。结论　SurvivinmRNA与蛋白在胃癌中表达较高 ,且与淋巴结转移、TNM分期、组织学分级有关 ,它可作为评估胃癌生物学行为和判断预后的生物学指标。     

青年乳腺癌患者P53的表达与临床病理特征的相关性分析

目的:探讨青年乳腺癌患者p53的表达与临床病理特征的相关性分析。方法:选取我院收治小于35岁乳腺癌患者35例为实验组,另选取35岁以上乳腺癌患者为对照组。比较两组患者病理组织分级、淋巴结转移数目以及P53的阳性表达率。结果:实验组P53阳性率为82.86%,高于对照组(65.71%),P0.05;P53阳性率随病理分级、淋巴结转移的增加而增高,经统计学分析呈正相关,P0.05。结论:青年乳腺癌患者的P53表达与病理组织学分级及淋巴结转移呈正相关,故p53的表达有助于判定青年乳腺癌患者的预后。     

COX-2、Ki-67和细菌L型感染在卵巢肿瘤中的表达及临床意义

目的探讨环氧合酶-2(COX-2)、Ki-67和细菌L型感染在卵巢肿瘤中的表达及临床相关性。方法应用免疫组化、原位杂交和革兰染色等方法检测了120例卵巢肿瘤中的COX-2、Ki-67蛋白及mRNA的表达,以及细菌L型的检出率。并对97例卵巢乳头状癌和23例卵巢乳头状瘤主要临床资料和病理分级参数进行比较,用χ2检验进行统计学处理。结果 COX-2、Ki-67蛋白及mRNA阳性表达恶性肿瘤明显高于良性肿瘤(P<0.01)。细菌L型检出阳性率与卵巢良、恶性肿瘤差异无统计学意义(P>0.5)。COX-2、Ki-67蛋白及mRNA阳性表达以及细菌L型检出阳性率与卵巢乳头状癌的临床分期、病理分级和腹腔淋巴结有转移有显著相关性(P<0.01)。细菌L型阳性患者中COX-2、Ki-67阳性明显高于L型阴性患者中阳性表达,2组差异具有统计学意义(P<0.01)。结论 COX-2、Ki-67蛋白及mRNA在卵巢肿瘤中有不同程度的异常表达,与卵巢癌的临床分期、病理分级和浸润、转移呈正相关,L型感染极有可能成为诱发肿瘤因素一,他们可能有协同致瘤作用。研究L型感染与卵巢肿瘤的关系,具有重要的临床应用价值。     

上皮性卵巢癌患者组织P53、Ki67表达及其临床意义

摘要 目的：探讨上皮性卵巢癌患者组织P53、Ki67表达及其临床意义。方法：选择卵巢肿瘤102例,其中病理诊断为良性卵巢肿瘤40例(良性组)和上皮性卵巢癌62例(恶性组),采用免疫组化法检测组织P53、Ki67表达水平,调查患者的临床病理特征并进行相关性分析。结果：恶性组的P53、Ki67表达阳性率为80.6 %和72.6 %,显著高于良性组的10.0 %和12.5 %(P<0.05)。在恶性组中,不同浸润转移、分化程度、病理分期患者的P53、Ki67表达阳性率对比差异有统计学意义(P<0.05)。直线相关分析显示上皮性卵巢癌患者P53表达阳性率与Ki67表达阳性率呈现显著正相关性(r=0.872,P=0.000)。多因素logistic回归分析显示浸润转移、分化程度、病理分期都为影响P53、Ki67表达阳性率的主要因素(P<0.05)。结论：上皮性卵巢癌患者组织P53、Ki67都呈现高表达状况,与患者的临床病理特征显著相关,两者也可互相影响,共同参与上皮性卵巢癌的发生与发展。     

LOXL4mRNA在膀胱癌中的表达及意义

膀胱癌是泌尿生殖系统最常见的恶性肿瘤,但其发生、发展的机制不清楚.通过采用逆转录聚合酶链反应(RT-PCR)方法检测58例膀胱癌组织、12例时照膀胱组织LOXL4(lysyl oxidase-like protein 4)mRNA的表达及其与临床分期、病理分级的关系.研究发现,58例膀胱癌组织中,LOXIA mRNA阳性表达率为24.1%(14/58),对照膀胱正常组织中LOXL4 mRNA阳性表达率为100%(12/12),膀胱癌组LOXL4 mRNA阳性表达率明显低于对照组(P<0.05).在膀胱癌组织不同临床分期中,Ta～1期阳性表达率为40%(10/25),T2～4期阳性表达率为12.1%(4/33),T2-4期膀胱癌组LOXL4 mRNA阳性表达率低于Ta～1期膀胱癌(P<0.05).不同病理分级膀胱癌组织中,G1阳性表达率为42.9%(9/21),G2～3阳性表达率为13.5%(5,37),G2～3膀胱癌组LOXIA mRNA阳性表达率低于G1膀胱癌(P<0.05).结果表明膀胱癌组LOXL4 mRNA表达水平明显低于正常对照组,LOXL4 mRNA的表达与膀胱癌的临床分期和病理分级呈负相关.提示LOXL4失表达、低表达可能是膀胱癌发生、发展的关键因素之一.     

Beclin1、p53与肺癌恶性度及疗效预后的相关性研究

目的通过定量检测肺组织中Beclin1和p53表达水平,分析自噬蛋白Beclin1与凋亡蛋白P53在肺癌发生发展中的作用,研究二者之间及其与肺癌临床病理分期的关系、为肺癌早期诊断提供新的思路和实验依据。方法选取外科手术或纤维支气管镜肺组织56例,依据2003年UICC公布的肺癌TNM分期标准同时结合临床表现、胸部CT、X线检查、纤维支气管镜检查、腹部CT或B超等将入选的50例病例进行分组:其中Ⅰ期5例,Ⅱ期10例,Ⅲ期(ⅢA+ⅢB)26例,Ⅳ期9例,各组年龄、性别等控制情况基本匹配。组织病理学分级按2003年UICC标准:G17例,G219例,G324例。另取6例癌旁组织或者病理确诊为正常组织作为对照组织。采用流式细胞术检测和分析不同病理分期以及不同临床分期肺癌组织中Beclin1、p53的表达水平,对该表达情况与对应的分期进行样本均数两两对比式统计学分析,并与正常肺组织作对照。结果 P53蛋白阳性表达百分率:肿瘤患者(63.96±9.43)%显著高于正常对照组(24.90±4.68)%,t=49.46,P0.05;肿瘤转移者显著高于未转移者,P0.05;并且P53蛋白表达随患者临床分期和病理分级的增加而逐渐增高(P0.05)。Beclin1蛋白检出率的变化规律与P53蛋白检出率的变化相反:肿瘤患者Beclin1蛋白(31.72±20.53)%,显著低于正常对照组(92.26±4.51)%,t=35.84,P0.05;该指标随患者临床分期和病理分级的增加而逐渐降低(P0.05)。Beclin1蛋白与P53蛋白二者相关性分析,Beclin1与p53的表达呈负相关,r=-0.848,P0.05。结论 Beclin1与p53的联合检测可以作为肺癌诊断指标,为估计肺癌的恶性度及预后提供依据。Beclin1与P53蛋白表达水平与肺癌发生发展的关系密切,肿瘤发生过程中细胞自噬作用降低和抗凋亡作用增强同时并存,提高自噬能力成为肿瘤治疗的又一途径。     

ATM mRNA在食管癌组织中的表达及其临床意义

目的探讨共济失调性毛细血管扩张症突变基因(Ataxia-telangiectasia mutated,ATM)mRNA在食管鳞状细胞癌组织中的表达及其临床意义。方法应用原位分子杂交方法检测52例食管正常黏膜、45例食管上皮内瘤变组织及63例食管癌组织ATM mRNA的表达。结果食管正常黏膜、食管上皮内瘤变及食管癌组织中ATM mRNA表达率分别为26.9%(14/52)、44.4%(20/45)及63.5%(40/63),食管癌组织中ATM mRNA表达率明显高于正常黏膜及上皮内瘤变(P≤0.05),ATM mRNA表达率与食管癌组织分级呈负相关(r=-0.312,P=0.013);食管正常黏膜和食管上皮内瘤变ATM mRNA表达率无明显差异(P=0.07),食管癌组织中ATM mRNA表达与患者年龄、性别、肿瘤浸润深度、淋巴结转移、临床分期及其它临床病理因素无关(P0.05)。结果 ATM mRNA在食管癌组织中异常表达,有望成为食管癌治疗的新靶点。     

非小细胞肺癌Survivin，Skp2和XIAP mRNA的表达及其临床意义

目的:探讨非小细胞肺癌(NSCLC)生存素(Survivin)、S期激酶相关蛋白2(Skp2)和X连锁凋亡抑制蛋白(XIAP)mRNA的表达及其临床意义。方法:收集2014年4月到2017年5月期间我院182例NSCLC患者在手术中切除的病理组织作为研究组,另收集每例患者癌变组织旁5cm以外的癌旁组织作为对照组。比较两组的Survivin、Skp2和XIAP mRNA阳性率,并分析Survivin、Skp2和XIAP mRNA表达与临床病理特征的关系。结果:研究组的Survivin、Skp2和XIAP mRNA阳性率显著高于对照组,差异有统计学意义(P0.05)。Survivin m RNA的阳性表达与年龄、性别、组织类型、分化程度、临床分期、淋巴结转移、吸烟史无关(P0.05)。Skp2 mRNA的阳性表达与年龄、性别、组织类型、分化程度、淋巴结转移、吸烟史无关(P0.05),与临床分期相关(P0.05)。XIAP mRNA的阳性表达与年龄、性别、组织类型、分化程度、吸烟史无关(P0.05),与临床分期、淋巴结转移相关(P0.05)。结论:Survivin、Skp2和XIAP m RNA在NSCLC患者的病理组织中呈高表达,Skp2 m RNA的阳性表达与临床分期有关,XIAP mRNA的阳性表达与临床分期、淋巴结转移有关。     

CAGE 基因mRNA水平表达与直肠癌关系的初步研究

目的:探讨直肠癌组织中癌相关基因(cancer-associated gene,CAGE)mRNA水平表达与直肠癌发生及临床分期、淋巴结转移的关系。方法:对我院2014年2月-2014年10月间肠镜室检查的标本,按病理检查结果分为3组:直肠癌组(80例;为直肠癌患者标本),大肠腺瘤组(38例;为大肠腺瘤患者标本),对照组(40例;为距肿瘤边缘10 cm以上标本)。采用RT-PCR法检测并比较各组CAGE在mRNA水平表达情况。甲基化特性PCR法(methylation-specific PCR,MSP)检测CAGE基因启动子的甲基化情况。将直肠癌患者分为CAGE基因甲基化患者组和CAGE基因去甲基化患者组,比较两组临床分期和淋巴结转移率。结果:与对照组相比,直肠癌组和大肠腺瘤组的CAGE mRNA水平表达明显升高(P0.01);直肠癌组高于大肠腺瘤组(P0.01)。3组间的基因启动子区的去甲基化阳性率差异有统计学意义(P0.01)。在直肠癌组中,CAGE去甲基化阳性患者的淋巴结转移率为73.8%(48/65),CAGE去甲基化阴性患者的淋巴结转移率为33.3%(5/15),两者相比差异有统计学意义(P0.01)。临床分期为I-Ⅱ期的CAGE基因启动子区的去甲基化直肠癌患者占24.6%(16/65),临床分期为Ⅲ-Ⅳ期的占75.4%(49/65);而甲基化的直肠癌患者中,临床分期为I-Ⅱ期的占66.7%(10/15),Ⅲ-Ⅳ期的占33.3%(5/15),两组间差异有统计学意义(P0.01)。结论:直肠癌组织和大肠腺瘤组织中,CAGE mRNA水平呈高表达。直肠癌的CAGE基因mRNA水平表达高于正常粘膜组织和大肠癌腺瘤组织。CAGE基因启动子区的去甲基化与淋巴结转移及临床分期密切相关。     

膀胱移行细胞癌中EphA2的表达和肿瘤MVD计数的关系

目的:探讨膀胱移行细胞癌中EphA2的生物学意义以及EphA2与微血管密度(MVD)的关系。方法:应用免疫组织化学技术检测85例膀胱移行细胞癌及10例正常膀胱粘膜中EphA2的表达,采用CD31染色标记微血管,行肿瘤微血管密度计数。结果:膀胱移行细胞癌和正常膀胱粘膜EphA2表达阳性率分别为89.4%(76/85)、40%(4/10),两组间差异有统计学意义(P<0.01);EphA2表达程度与膀胱癌的病理分级、临床分期和淋巴结转移均有相关性(P<0.05);膀胱移行细胞癌EphA2阴性组与EphA2阳性纽间肿瘤微血管密度(MVD)计数差异有统计学意义(P<0.05),EphA2不同阳性程度组之间MVD计数差异没有统计学意义(P>0.05)。结论:EphA2可作为判定膀胱癌恶性程度的参考指标,与肿瘤微血管生成相关,有望成为膀胱癌治疗的新靶向。     

HuR Is Necessary for Mammary Epithelial Cell Proliferation and Polarity at Least in Part via ΔNp63

HuR, a RNA binding protein, is known to function as a tumor maintenance gene in breast cancer and associated with tumor growth and poor prognosis. However, the cellular function of this protein remains largely unknown in normal mammary epithelial cells. Here, we showed that in immortalized MCF10A mammary epithelial cells, HuR knockdown inhibits cell proliferation and enhances premature senescence. We also showed that in three-dimensional culture, MCF10A cells with HuR knockdown form abnormal acini with filled lumen and an aberrant expression pattern of the extracellular matrix protein laminin V. In addition, we showed that HuR knockdown increases ΔNp63, but decreases wild-type p53, expression in MCF10A cells. Moreover, we showed that ΔNp63 knockdown partially rescues the proliferative defect induced by HuR knockdown in MCF10A cells. Consistent with this, we identified two U-rich elements in the 3′-untranslated region of p63 mRNA, to which HuR specifically binds. Finally, we showed that HuR knockdown enhances ΔNp63 mRNA translation but has no effect on p63 mRNA turnover. Together, our data suggest that HuR maintains cell proliferation and polarity of mammary epithelial cells at least in part via ΔNp63.     

EGFR through STAT3 modulates ΔN63α expression to sustain tumor‐initiating cell proliferation in squamous cell carcinomas

Many squamous cell carcinomas (SCCs) are characterized by high levels of EGFR and by overexpression of the ΔNp63α isoform. Here, we investigated the regulation of ΔNp63α expression upon EGFR activation and the role of the EGFR–ΔNp63α axis in proliferation of SCC tumor‐initiating cells (TICs). SCC cell lines A‐431, Cal‐27, and SCC‐25 treated with EGF showed a time‐dependent increase in ΔNp63α expression at the protein and mRNA levels, which was blocked by the tyrosine kinase inhibitor (TKI) Lapatinib. RNA interference experiments suggested the role of STAT3 in regulating ΔNp63α expression downstream of EGFR. Inactivation of EGFR by the monoclonal antibody Cetuximab and RNA interference against STAT3 or ΔNp63α impaired the TICs ability to grow under non‐differentiating conditions. Radiation treatment, which triggers EGFR activation, induced ΔNp63α accumulation without affecting TICs proliferation, whereas the combination Cetuximab plus radiation significantly reduced TICs growth under non‐differentiating conditions. Together, our findings provide evidence that ΔNp63α expression is regulated by EGFR activation through STAT3 and that the EGFR–ΔNp63α axis is crucial for proliferation of TICs present in SCCs. J. Cell. Physiol. 228: 871–878, 2013. © 2012 Wiley Periodicals, Inc.     

Delta Np63 alpha expression is regulated by the phosphoinositide 3-kinase pathway

p63 is a homologue of p53 that functions to maintain progenitor cell populations in stratified epithelia. Delta Np63 alpha is overexpressed in epithelial cancers and has been shown to have oncogenic properties. We have previously reported that inhibition of epidermal growth factor receptor signaling results in a decrease in Delta Np63 alpha expression. Here, we demonstrate Delta Np63 alpha is a target of the phosphoinositide-3-kinase (PI3K) pathway downstream of the epidermal growth factor receptor. Treatment of keratinocytes with epidermal growth factor results in an increase in Delta Np63 alpha expression at the mRNA level, which is abrogated by inhibition of PI3K but not mitogen-activated protein kinase signaling. Small interfering RNA-mediated knockdown of the p110 beta catalytic subunit of PI3K results in a decrease in Delta Np63 alpha protein levels in keratinocytes. The results presented herein suggest that regulation of Delta Np63 alpha expression by the PI3K pathway plays a critical role in the survival and proliferative capacity of squamous epithelia.     

ΔNp63α regulates keratinocyte proliferation by controlling PTEN expression and localization

ΔNp63α, implicated as an oncogene, is upregulated by activated Akt, part of a well-known cell survival pathway. Inhibition of Akt activation by phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and the presence of putative p63-binding sites in the pten promoter led us to investigate whether ΔNp63α regulates PTEN expression. Knockdown of ΔNp63α led to increases in PTEN levels and loss of activated Akt, while overexpression of ΔNp63α decreased PTEN levels and elevated active Akt. The repression of PTEN by ΔNp63α occurs independently of p53 status, as loss of ΔNp63α increases PTEN expression in cell lines with and without functional p53. In addition, decreased levels of ΔNp63α resulted in an increase in nuclear PTEN. Conversely, in vivo nuclear PTEN was absent in the proliferative basal layer of the epidermis where ΔNp63α expression is highest. Additionally, we show that in keratinocytes a balance between ΔNp63α and PTEN regulates Akt activation and maintains normal proliferation rates. This balance is disrupted in non-melanoma skin cancers through increased ΔNp63α levels, and could enhance proliferation and subsequent neoplastic development. Our studies show that ΔNp63α negatively regulates PTEN, thereby providing a feedback loop between PTEN, Akt and ΔNp63α, which has an integral role in skin cancer development.     

Evaluation of the Content of Antimony,Arsenic, Bismuth,Selenium, Tellurium and Their Inorganic Forms in Commercially Baby Foods

The purpose of the investigation is to reveal the influence of dietary calcium on fluorosis-induced brain cell apoptosis in rat offspring, as well as the underlying molecular mechanism. Sprague–Dawley (SD) female rats were randomly divided into five groups: control group, fluoride group, low calcium, low calcium fluoride group, and high calcium fluoride group. SD male rats were used for breeding only. After 3 months, male and female rats were mated in a 1:1 ratio. Subsequently, 18-day-old gestation rats and 14- and 28-day-old rats were used as experimental subjects. We determined the blood/urine fluoride, the blood/urine calcium, the apoptosis in the hippocampus, and the expression levels of apoptosis-related genes, namely Bcl-2, caspase 12, and JNK. Blood or blood/urine fluoride levels and apoptotic cells were found significantly increased in fluorosis rat offspring as compared to controls. Furthermore, the Bcl-2 messenger RNA (mRNA) expression levels significantly decreased, and caspase 12 mRNA levels significantly increased in each age group as compared to controls. Compared with the fluoride group, the blood/urine fluoride content and apoptotic cells evidently decreased in the high calcium fluoride group, Bcl-2 mRNA expression significantly increased and caspase 12 mRNA expression significantly decreased in each age group. All results showed no gender difference. Based on these results, the molecular mechanisms of fluorosis-induced brain cell apoptosis in rat offspring may include the decrease in Bcl-2 mRNA expression level and increase in caspase 12 mRNA expression signaling pathways. High calcium intake could reverse these gene expression trends. By contrast, low calcium intake intensified the toxic effects of fluoride on brain cells.