鼻咽癌中IGF-1R、p-ERK、c-fos和Ki-67蛋白的表达及意义

目的研究胰岛素样生长因子1受体（IGF-1R）蛋白与细胞增殖相关信号分子p-ERK、c-fos、Ki-67蛋白在鼻咽癌及鼻咽黏膜上皮细胞中的表达及意义。方法 SP法检测30例鼻咽癌、25例鼻咽黏膜上皮细胞中IGF-1R、p-ERK、c-fos、Ki-67蛋白的表达。结果 IGF-1R、p-ERK、c-fos和Ki-67蛋白在鼻咽癌细胞中阳性表达率分别为93.33%（28/30）、93.33%（28/30）、83.33%（25/30）、86.67%（26/30）,在鼻咽黏膜上皮细胞中阳性表达率分别为36.00%（9/25）、28.00%（7/25）、20.00%（5/25）、4.00%（1/25）,2组间差异有显著统计学意义（P〈0.01）;鼻咽癌中IGF-1R与p-ERK、c-fos、Ki-67蛋白阳性表达呈正相关（P〈0.05）;p-ERK与c-fos、Ki-67蛋白阳性表达呈正相关（P〈0.01）;c-fos和Ki-67蛋白阳性表达呈正相关（P〈0.01）。结论鼻咽癌中IGF-1R蛋白过表达参与调控细胞增殖。     

下调VEGF表达影响鼻咽癌细胞放射敏感度的机制

目的　应用RNA干扰技术抑制人鼻咽低分化鳞状上皮细胞癌细胞株CNE-2中血管内皮生长因 子（VEGF）表达,研究阻断VEGF基因表达对鼻咽癌细胞放射敏感度的影响及机制。方法 构建针对 VEGF的siRNA真核表达载体pU-VEGF-siRNA（CNE-2组）、1个阴性对照质粒（CNE-2/Neg-siRNA 组）、经脂质体转染至CNE-2细胞（CNE-2/VEGF-siRNA组）,用平板克隆形成实验检测在6MV-X线 0、2、4、6、8、10 Gy照射后克隆形成能力及通过单击多靶模型、线性二次模型拟合放射生物学参 数,流式细胞检测分析细胞周期和细胞凋亡的变化,用RT-PCR定量分析三组细胞中Cyclin D1、Cyclin E、P16和P53的mRNA表达。结果 经6MV-X线0、2、4、6、8、10 Gy照射后细胞存活率显著下降, CNE-2/VEGF-siRNA组细胞经D0和2 Gy照射后的存活分数明显低于CNE-2组、CNE-2/Neg-siRNA组; CNE-2/VEGF-siRNA组中G1/S期细胞周期阻滞更为明显。在Cyclin D1、Cyclin E、p16和p53基因中, Cyclin D1mRNA表达在放疗后6、12和24 h进行性升高,差异有统计学意义,而其他基因变化差异无统 计学意义,Western blot检测显示Cyclin D1蛋白表达在放疗后24 h明显升高。结论 下调VEGF表达可 增加鼻咽癌细胞的放射敏感度,其机制可能是通过Cyclin D1信号通路途径使细胞周期发生G1/S期阻滞。     

lncRNA LAMA5-AS1通过上调TFPI2表达抑制鼻咽癌细胞的增殖和迁移

目的:检测鼻咽癌组织和细胞株中长链非编码RNA（long non-coding RNA，lncRNA）LAMA5-AS1的表达，观察lncRNA LAMA5-AS1对鼻咽癌细胞增殖和迁移的影响并探讨其作用机制。方法:荧光实时定量聚合酶链式反应（qPCR）分别检测lncRNA LAMA5-AS1在68例鼻咽癌组织及62例慢性鼻咽炎组织、鼻咽癌细胞株及永生化鼻咽上皮细胞中的相对表达。选择lncRNA LAMA5-AS1表达最少的细胞株，分别转染阴性对照质粒（对照组）或载有lncRNA LAMA5-AS1序列质粒（实验组）。MTS法和细胞划痕实验分别检测高表达lncRNA LAMA5-AS1对细胞增殖和迁移能力的影响。qPCR检测高表达lncRNA LAMA5-AS1对组织因子途径抑制物2（tissue factor pathway inhibitor 2，TFPI2）基因mRNA表达的影响，Western blotting检测TFPI2蛋白和ERK信号通路蛋白表达。结果:与慢性鼻咽炎组织比较，lncRNA LAMA5-AS1在鼻咽癌组织中表达明显降低（P＜0.01）。与人永生化鼻咽上皮细胞比较，lncRNA LAMA5-AS1在鼻咽癌细胞株中表达明显降低（P＜0.01），在CNE-2Z细胞中的表达最少（P＜0.01）。与对照组比较，实验组CNE-2Z细胞的增殖能力从第3天开始明显降低（P＜0.05），实验组CNE-2Z细胞迁移能力明显下降（P＜0.01）。与对照组比较，实验组CNE-2Z细胞中TFPI2在mRNA和蛋白水平的表达明显增加（P＜0.01），ERK信号通路蛋白p-ERK、MSK1、MNK和STAT3表达明显降低。结论:lncRNA LAMA5-AS1在鼻咽癌组织和细胞株中表达明显降低，高表达lncRNA LAMA5-AS1可通过上调TFPI2基因表达、下调ERK信号通路活性，抑制鼻咽癌CNE-2Z细胞的增殖和迁移能力。     

SHP-1/p21/CDK6/Cyclin D1在不同放射敏感度鼻咽癌细胞中的表达

目的 建立鼻咽癌放射抗拒亚系CNE-2S,研究SHP-1/p21/CDK6/Cyclin D1通路在鼻咽癌细胞系CNE-2和CNE-2S中表达的差异,探讨鼻咽癌放射敏感度变化的分子机制.方法 通过X线大剂量低分割照射技术建立鼻咽癌放射抗拒亚系(子代,CNE-2S1),观察亲代(CNE-2)和子代细胞的形态学和生长动力学差异,检测亲代及子代细胞系放射敏感度相关参数,分析亲代及子代细胞系各自细胞周期分布,同时检测亲代与子代细胞系SHP-1/p21/CDK6/Cyclin D1通路中各因子的蛋白质表达水平.结果 通过X线大剂量低分割照射技术成功建立鼻咽癌放射抗拒亚系CNE-2S1,且CNE 2及CNE-2S1放射敏感度相关参数具有延续性.同时CNE-2S1细胞S期比例较CNE-2细胞明显增高,G1期细胞明显减少,G2-M期细胞比例变化不明显.此外SHP-1、CDK6以及CylinD1在CNE-2S1细胞中的蛋白表达水平明显上调,p21表达水平则明显下调,其与CNE-2细胞之间的差异具有统计学意义.结论 SHP-1/p21/CDK6/Cyclin D1通路可能在调控鼻咽癌放射敏感度和细胞周期分布方面发挥一定作用.     

ERK途径在胰岛素促进子宫内膜癌细胞增殖中的作用

背景与目的 观察胰岛素对子宫内膜癌细胞ERK信号通路的活化,并探讨ERK信号通路对Cyclin D1mRNA、蛋白水平的影响及该通路在细胞增殖调节中的作用.方法 将无血清饥饿的子宫内膜癌Ishikawa3-H-12细胞分为空白对照组、10-6mol/L胰岛素单独刺激组以及不同剂量MEK抑制剂PD98059预处理后再用胰岛素刺激组.Western blot检测各组ERK磷酸化(P-ERX)及Cyclin D1蛋白水平.RT-PCR检测Cyclin D1 mRNA水平,MTT试验观察细胞增殖情况.结果 胰岛素可引起子宫内膜癌细胞ERK活化,刺激30 min后P-ERK/ERK比值显著高于空白对照组(65.419% vs22.45%,P<0.001),胰岛素可促进子宫内膜癌细胞Cyclin D1 mRNA转录和蛋白表达,分别刺激12h、24h后Cyclin D1mRNA转录和蛋白表达水平显著高于空白对照组(分别为84.62% vs 18.55%;74.04% vs 32.91%,均P<0.001),PD98059以浓度依赖方式抑制胰岛素引起的ERK磷酸化、mRNA转录和蛋白表达水平.MTT试验显示,在药物处理24 h,48 h和72 h 3个时间点,不同组570 nm吸光度值(OD570)均有显著差异(F=204.160;33.850;90.764,均P<0.001).胰岛素组OD570值均高于同时间点的空白对照组(均P<0.001),PD98059可抑制胰岛素的增殖促进作用,且具有浓度依赖性.结论 胰岛素可以经由ERK(途径促进Cydin D1 nRNA车专录和蛋白表达,并进一步促进细胞增殖.     

PD98059抑制MAPK/ERK信号通路对胃癌细胞生物学功能的影响

目的：研究MAPK/ERK信号通路中关键信号分子MEK和ERK在胃癌SGC-7901细胞中的表达及PD98059抑制MAPK/ERK通路对胃癌细胞生物学功能的影响。方法：体外培养胃癌细胞株SGC-7901,不同浓度（0、25、50、100、200、300和400 mmol/L）PD98059处理24 h后CCK-8法检测细胞增殖率变化;再用0、25、50和100 μmol/L PD98059处理24 h后采用实时荧光定量PCR（qPCR）检测MEK和ERK mRNA的表达量;Western blot检测MEK和ERK蛋白的表达;流式细胞术检测细胞周期和凋亡变化。同时设正常胃黏膜上皮GES-1细胞为对照。结果：与正常胃黏膜上皮GES-1细胞相比,胃癌SGC-7901细胞中MEK和ERK mRNA的表达升高,差异具有统计学意义（P < 0.05）;p-MEK、p-ERK蛋白的表达亦显著升高,差异具有统计学意义（P < 0.05）。0~200 μmol/L PD98059处理SGC-7901细胞后,细胞增殖率随着抑制剂浓度的升高而降低（P < 0.05）。当PD98059浓度处于200~400 μmol/L时抑制作用逐渐趋于平稳。0~100 μmol/L PD98059作用后MEK、ERK mRNA的表达量低于对照组（P < 0.05）,随着PD98059浓度升高,ERK mRNA表达量逐渐降低（P < 0.05）。Western blot检测结果显示50和100 μmol/L PD98059作用后p-MEK1/2、p-ERK1/2蛋白表达降低（P < 0.05）。且抑制剂PD98059使胃癌SGC-7901细胞发生G0/G1期阻滞,可诱导细胞凋亡。结论：MAPK/ERK信号通路在胃癌细胞中激活,PD98059通过抑制MAPK/ERK信号通路的活性可影响胃癌细胞的生物学功能。     

姜黄素对人鼻咽癌CNE-2Z细胞增殖及 凋亡的影响

 目的探讨姜黄素对人鼻咽癌CNE-2Z细胞增殖及凋亡的影响及其相关的分子机制。方法体外培养的 CNE-2Z细胞经不同剂量姜黄素处理后,用MTT法观察对细胞增殖的影响,流式细胞术检测细胞周期和凋亡 率的改变,Western blot检测姜黄素处理后相关蛋白Bcl-2、Bax表达水平的变化。结果姜黄素对CNE-2Z细 胞的生长和增殖都具有一定程度的抑制作用,阻断细胞于S期和G2/M期。姜黄素处理后CNE-2Z细胞凋亡抑 制基因Bcl-2蛋白表达减少,同时促凋亡基因Bax蛋白表达增加。 结论姜黄素可抑制CNE-2Z细胞增殖并促进 其凋亡。     

Cyclin D1、Ki-67在宫颈鳞状细胞癌中的表达及临床意义

目的 探索细胞周期蛋白Cyclin D1和细胞增殖活性标记物Ki-67在宫颈鳞状细胞癌中的表达及其与临床病理特征的相关性.方法 采用免疫组织化学方法检测宫颈鳞状细胞癌中Cyclin D1和Ki-67的表达，并进行相关分析。结果 Cyclin D1在宫颈鳞状细胞癌组织中阳性率为46．7％（14／30）；Ki-67在宫颈鳞状细胞癌组织中阳性率为73．3％（22／30）。Cyclin D1表达与有无转移呈正相关（P〈0．05），与病理分级及临床分期无明显相关性（P〉0．05）。Ki-67表达与病理分级及临床分期呈正相关（P〈0．05），与有无转移无明显相关性（P〉0．05）。结论 Cyclin D1和Ki-67的过表达可能与宫颈鳞状细胞癌的高侵袭性和不良预后密切相关。     

PNCK对鼻咽癌细胞增殖和凋亡作用的研究

目的 研究妊娠上调的非泛素性钙调蛋白激酶(Pregnancy-upregulated nonubiquitous calmodulin kinase,PNCK)在鼻咽癌组织中的表达以及对鼻咽癌细胞增殖和凋亡的作用。方法 采用免疫组织化学方法检测鼻咽癌癌组织和正常组织中PNCK表达水平。构建慢病毒载体干扰PNCK基因表达,并采用Real time PCR和免疫印迹结果证实,采用MTT方法、流式细胞术、Caspase3和Caspase7活性检测鼻咽癌肿瘤细胞增殖和凋亡的变化。结果 免疫组织化学结果显示PNCK蛋白在鼻咽癌癌组织中呈特异性高表达。Real time PCR和免疫印迹结果证实慢病毒成功干扰鼻咽癌CNE-2Z细胞中PNCK表达(PNCK基因在mRNA和蛋白水平表达受到显著抑制)。MTT检测结果显示抑制PNCK表达抑制CNE-2Z细胞增殖。此外,干扰PNCK表达引起CNE-2Z细胞Caspase3和Caspase7活性上升,流式细胞术表明干扰PNCK基因表达可以促进CNE-2Z细胞凋亡。结论 干扰PNCK基因表达抑制鼻咽癌细胞增殖并诱导肿瘤细胞凋亡,可能是治疗鼻咽癌的潜在靶点。     

整合素α5β1和细胞外信号调节激酶信号传导通路在非小细胞肺癌中的作用及其相关性研究

研究整合素α5β1 及磷酸化ERK1/2 （p-ERK1/2） 在非小细胞肺癌（NSCLC）组织中的表达及整合素α5β1 和ERK1/2 信号通路在人肺癌A549 细胞生长和迁移中的作用。方法:采用免疫组织化学法和免疫印迹（Western blot）法检测41 例NSCLC 组织和15例正常肺组织中整合素α5β1 及p-ERK1/2 的表达并分析两者的相关性,体外培养肺癌A549 细胞,以Anti-α5β1 或ERK 激酶抑制剂PD98059 作为工具药物,采用四甲基偶氮唑盐比色 （MTT） 法和 Annexin-V FITC PI 双染色法检测A549 增殖和凋亡,采用Western blot 检测A549 中MMP-9 蛋白水平。采用逆转录-聚合酶链反应（RT-PCR）和Western blot 检测Anti-α5β1 处理后A549 中ERK1/ 2 蛋白和mRNA 表达水平的变化。结果:人NSCLC 组织中整合素α5β1 及p-ERK1/2 阳性表达率（分别为58.53%,65.52%）明显高于正常肺组织（分别为26.67%, 20.00%）（P<0.01）。整合素α5β1与p-ERK1/2 表达均与病理分级（分别为P<0.001,P=0.030）、淋巴结转移（分别为P=0.014,P<0.001）、临床分期（分别为P=0.027,P=0.038）相关。整合素α5β1 与p-ERK1/2表达具有相关性（r=0.375, P<0.05）。经Anti-α5β1 或PD98059 处理后,A549 细胞增殖效应和早期凋亡细胞百分率增加,MMP-9 的表达均明显减弱。经Anti-α5β1 处理后A549 细胞中ERK 的mRNA 和蛋白表达受到抑制,ERK1/2 的磷酸化水平显著减少。结论: 整合素α5β1 及p-ERK1/2 在人NSCLC 组织中高表达,Anti-α5β1 在下调了细胞内ERK 的mRNA 和蛋白磷酸化的同时,可以明显抑制A549 细胞的增殖和MMP-9 蛋白表达,表明整合素α5β1 可能介导ERK1/2 信号转导通路参与了非小细胞肺癌中的异常增殖和迁移的调控。      

Bacteria and cancer--antagonisms and benefits

There is considerable historical and recent evidence concerning the antagonisms between acute bacterial infections or their toxins and cancer and allied diseases. These data provide renewed incentives to undertake clinical programmes with mixed bacterial vaccines in many countries at the present time.     

VEGF及KDR在大肠腺瘤和腺癌中的表达及意义

目的：探讨VEGF和KDR在大肠腺瘤和大肠腺癌中的表达及临床病理特征的关系。方法：大肠腺瘤和大肠腺癌组织标本各100例,采用免疫组织化学染色法检测VEGF和KDR在标本中的表达情况。结果：VEGF和KDR在大肠腺癌组中的阳性表达明显高于大肠腺瘤组（P〈0.05）;在正常大肠黏膜均未见VEGF和KDR表达的阳性染色;VEGF阳性表达组中KDR的阳性表达率为70%,显著高于VEGF阴性表达组中KDR的阳性表达率16%,两组比较有统计学意义（P〈0.01）。结论：大肠腺癌组织中KDR的表达与肿瘤大小、转移情况、浸润深度密切相关;VEGF和KDR在大肠腺瘤中的表达与患者的年龄、性别及分型均无相关性,而与增生程度相关（P〈0.05）。在大肠腺癌患者中VEGF及KDR表达更高,二者具有协同效应。     

Religiosity and physical and emotional functioning among African American and White colorectal and lung cancer patients

The literature suggests that religiosity helps cope with illness. The present study examined the role of religiosity in functioning among African Americans and Whites with a cancer diagnosis. Patients were recruited from an existing study and mailed a religiosity survey. Participants (N = 269; 36% African American, 56% women) completed the mail survey, and interview data from the larger cohort was utilized in the analysis. Multivariate analyses indicated that in the overall sample religious behaviors were marginally and positively associated with mental health and negatively with depressive symptoms. Among women, religious behaviors were positively associated with mental health and negatively with depressive symptoms. Religiosity was not a predictor of study outcomes for men. Among African Americans, religious behaviors were positively associated with mental health and vitality. Among Whites, religious behaviors were negatively associated with depressive symptoms. These findings suggest a mixed role of religious involvement in cancer outcomes. The current findings may have applied potential in the areas of emotional functioning and depression.     

Genetic polymorphisms of alcohol and aldehyde dehydrogenases and risk for esophageal and head and neck cancers

Alcoholic beverages are causally related to cancer of the oral cavity, pharynx, larynx and esophagus. Ethanol is oxidized to acetaldehyde and then to acetate by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), both of which have genetic polymorphisms. A review of case-control studies of the effects of ALDH2, ADH2 and ADH3 genotypes shows consistently positive associations between inactive heterozygous ALDH2 and the less-active ADH2 genotypes and the risk for esophageal cancer in East Asian heavy drinkers and this enzyme-related vulnerability may extend to light-to-moderate drinkers. Some studies suggest similar associations with the risk for head and neck cancer in moderate-to-heavy-drinking Japanese. An established carcinogen in experimental animals, acetaldehyde can interact with human DNA. ALDH2-associated cancer susceptibility fits into a scenario in which acetaldehyde plays a critical role in the development of human cancer. Alcohol flushing and drinking behavior may partly explain this carcinogenic effect in carriers of less-active ADH2 genotypes. Whether the ADH3 genotype influences head and neck cancer risk in Western nations is controversial. Professional and public education about risky conditions connected to the ALDH2 and ADH2 genotypes and environmental factors is important in a new strategic approach to the prevention of alcohol-related cancers in East Asians. The use of simple tests to identify inactive ALDH2 on the basis of alcohol flushing responses could benefit many people, by helping them to identify their own cancer risks. Such testing could also help clinicians diagnose esophageal cancer earlier, through the use of endoscopic screening in the high-risk population.     

Religiosity and Physical and Emotional Functioning Among African American and White Colorectal and Lung Cancer Patients

The literature suggests that religiosity helps cope with illness. The present study examined the role of religiosity in functioning among African Americans and Whites with a cancer diagnosis. Patients were recruited from an existing study and mailed a religiosity survey. Participants (N = 269; 36% African American, 56% women) completed the mail survey, and interview data from the larger cohort was utilized in the analysis. Multivariate analyses indicated that in the overall sample religious behaviors were marginally and positively associated with mental health and negatively with depressive symptoms. Among women, religious behaviors were positively associated with mental health and negatively with depressive symptoms. Religiosity was not a predictor of study outcomes for men. Among African Americans, religious behaviors were positively associated with mental health and vitality. Among Whites, religious behaviors were negatively associated with depressive symptoms. These findings suggest a mixed role of religious involvement in cancer outcomes. The current findings may have applied potential in the areas of emotional functioning and depression.     

New and emerging radiosensitizers and radioprotectors

The combination of chemotherapy and radiation has led to clinical breakthroughs in several disease sites, and current work continues to define optimum combinations of proven chemotherapy as well as more recently available, noncytotoxic agents. Administration of systemic therapies allows modulation of radiation response to improve tumor control (radiosensitization) or to prevent normal tissue toxicity (radioprotection). Substantial progress has been made in identifying the targets of standard chemotherapeutic radiation sensitizers and protectors as well as in the introduction of a new generation of molecularly targeted therapies in combination with radiation. We have reviewed the most recent, predominantly early phase clinical trials combining systemic agents with radiation. Although the proof of an improved schedule ultimately needs to come from well-run Phase III trials, the search among schedules could be shortened by the use of surrogate endpoints such as presence of active drug metabolites in the tumor. This has been accomplished only in a few cases and needs to become a more standard part of radiation sensitizer and protector trials.