西妥昔单抗对人鼻咽癌细胞抑制作用的体外研究

目的:探讨西妥昔单抗(C225)以不同序贯方式联合顺铂(DDP)对人鼻咽癌HNE1亲代和顺铂耐药细胞株HNE1/CDDP的生长抑制作用.方法:以人鼻咽癌HNE1亲代和DDP耐药细胞株HNE1/CDDP作为研究对象.免疫细胞化学染色法检测HNE1和HNE1/CDDP细胞EGFR蛋白的表达;MTT法和克隆形成法检测C225单药或与DDP联合对HNE1和HNE1/CDDP细胞的生长抑制作用.结果:1)HNE1和HNE1/CDDP细胞EGFR的阳性率分别为(87.75±5.44)%和(89.63±4.89)%,差异无统计学意义,t=0.724,P=0.481.2)在1～200 μg /mL范围内,C225对HNE1和HNE1/CDDP细胞的最大生长抑率分别为(28.17±3.47)%和(17.65±1.73)%,其生长抑制作用均不呈剂量依赖性,P＞0.15.3)10 μg/mL C225与DDP联合作用于HNE1和HNE1/CDDP细胞,无论先C225后DDP还是先DDP后C225的序贯联合用药方式,均呈现相加作用.结论:C225单药对鼻咽癌HNE1亲代和顺铂耐药细胞株HNE1/CDDP有较温和的生长抑制作用,与DDP联合时不受序贯顺序影响,均呈现相加作用.     

人鼻咽癌耐药细胞株的建立及其与LRP的关系

建立人鼻咽癌多药耐药细胞株TW03/DDP并研究其与肺耐药相关蛋白（LRP）的关系。方法：以顺铂（DDP）为诱导药物,诱导人鼻咽癌细胞TW03,建立鼻咽癌多药耐药细胞株TW03/DDP;MTT法检测药物敏感性,流式细胞术检测细胞周期的改变,并绘制细胞生长曲线、测定倍增时间,观察细胞形态学变化;通过免疫细胞化学法、免疫印迹法、RT-PCR检测LRP的表达。结果：TW03/DDP对顺铂、5-氟尿嘧啶、阿霉素、阿糖胞苷、长春新碱等药物的耐药指数分别是40.85、5.65、26.68、1.60与28.19,其半数抑制浓度与TW03比较差异有统计学意义（P<0.05）;耐药细胞株的细胞倍增时间相对于其亲本细胞略有延长,其G0/G1期明显减少而G2/M期则相对增加;免疫细胞化学、免疫印迹法、RT-PCR检测显示LRP及其mRNA在TW03/DDP中的表达明显高于TW03,差异具有统计学意义（P<0.05）。结论：本研究建立了人鼻咽癌多药耐药细胞株TW03/DDP,其LRP在蛋白水平及mRNA水平的表达高于亲本细胞,表明LRP的表达上调与其耐药机制有密切关系。     

人鼻咽癌耐顺铂细胞系的放射增敏实验

目的:了解人鼻咽低分化鳞癌细胞株HNE1及其耐药株HNE1/DDP的放射敏感性,探讨顺铂(DDP)对耐药株及其亲本细胞株放射增敏的差别,并初步研究吉西他滨对HNE1/DDP的放射增敏作用及其机制.方法:实验分为单纯照射组和照射加药组,应用克隆形成方法,观察单纯照射和照射加药物(DDP或吉西他滨)对细胞的杀伤作用,用Radiomed软件进行曲线拟合作图.免疫组织化学、末端脱氧核苷酰转移酶法(TUNEL法)和流式细胞法观察吉西他滨对耐药细胞株的影响.结果:DDP对HNE1和HNE1/DDP的放射增敏比分别为1.201 5和0.941 8,而吉西他滨对HNE1/DDP的放射增敏比为1.379 1.吉西他滨作用后耐药细胞凋亡增加,bax表达上调,p53没有明显变化,药物作用前后Bcl-2表达均呈阴性,并且使肿瘤细胞周期阻滞在G1期,而S期细胞比例则明显减少.结论:鼻咽低分化鳞癌细胞株HNE1对DDP和放射线没有交叉抵抗.DDP对HNE1具有放射增敏作用,但是对HNE1/DDP无放射增敏作用,而吉西他滨对HNE1/DDP有放射增敏作用,其增敏机制可能与吉西他滨使细胞周期阻滞在G1/S期,以及细胞p53、bax和Bcl-2基因表达变化导致凋亡增加有关.     

鼻咽癌相关基因NAG7对鼻咽癌细胞周期及凋亡的影响

背景与目的NAG7基因是我室克隆的鼻咽癌相关的肿瘤抑制候选基因,其功能与作用机制目前尚不清楚.研究鼻咽癌相关的潜在抑瘤基因NAG7对鼻咽癌细胞系(HNE1)细胞周期和细胞凋亡的影响,并探讨其作用机制.方法采用脂质体转染技术将NAG7基因导入HNE1细胞,建立稳定表达NAG7的细胞株,Northernblot分析转染细胞NAG7基因的表达,并采用流式细胞技术检测细胞周期、细胞周期素及细胞凋亡的改变,并用westernblot验证.结果NAG7基因重表达的HNE1细胞与空载体转染的HNE1和HNE1细胞相比G0/G1期细胞数增加(P<0.05),S期细胞数减少;细胞凋亡数目增加(P<0.05);细胞周期素A、D1、E表达明显降低(P<0.05),细胞周期素B1表达降低,Westernblot检测亦证实cyclinD1和cyclinE表达明显下调.结论NAG7的重表达导致细胞周期素表达下调,从而延缓细胞经G1期进入S周期及诱导细胞凋亡增加进而抑制NPC细胞的过度增殖.     

高温合并化疗对人胃癌细胞株增殖及凋亡影响的观察

目的:观察高温合并化疗对人胃癌细胞的细胞周期和细胞凋亡的影响,了解高温逆转肿瘤多药耐药性和高温与化疗协同作用的分子生物学机制.方法:以体外培养人胃癌细胞株SGC7901/ADM为研究对象,选用37、40、43℃3种不同温度,以及ADM、DDP和TAX 3种不同药物.实验分为单纯温度组、单纯药物组、不同温度+不同化疗药物组.采用PI染色法,利用流式细胞仪测定细胞周期和细胞凋亡率.结果:高温下处于S期的细胞数量明显减少,进入G0～G1期细胞增多,而细胞凋亡率仅增加2.3％～6.3％,P=0.001 7.高温联合化疗时,细胞凋亡的数量显著增加.ADM组43和40℃时细胞凋亡率分别为(65.7±2.9)％和(50.8±1.7)％,与37℃的细胞凋亡率(36.2±1.3)％相比,差异有统计学意义,P=0.001.DDP组40和43℃下,细胞凋亡率显著提高到(42.5±0.9)％和(58.4±3.2)％,与37℃的细胞凋亡率(24.4±0.6)％相比差异有统计学意义,P=0.001.TAX组40和43℃时,细胞凋亡率升至(38.2±2.3)％和(40.5±4.2)％,与37℃的细胞凋亡率(11.2士0.7)％相比差异有统计学意义,P=0.001.结论:高温增加耐药细胞对药物的敏感性,可通过影响细胞周期,诱导细胞凋亡所产生.     

单纯加温与加温联用顺铂对人卵巢癌细胞增殖及凋亡的影响

目的探讨单纯加温与联用顺铂(DDP)对卵巢癌细胞增殖及凋亡的影响.方法采用MTT法检测加温及加温联合DDP对卵巢癌DDP敏感细胞系skov3和DDP耐药细胞系SK-OV-3细胞增殖的抑制效应,分析加温与DDP在抑制卵巢癌细胞增殖中的相互作用;采用TUNEL和流式细胞术分析加温及加温联合DDP对卵巢癌细胞周期和凋亡的影响,探讨热化疗的作用机制.结果 42℃以上加温可显著抑制卵巢癌DDP敏感和耐药细胞系细胞增殖(P<0.001),且与加热温度和时间之间存在剂量效应关系.加热与DDP在抑制卵巢癌细胞增殖上表现为相加或协同作用,在skov3细胞系以相加作用(0.85≤q≤1.15)为主,而在SK-OV-3细胞系则主要表现为协同作用(q>1.15).加温可诱导卵巢癌细胞凋亡,并降低S期细胞比率和增加G0/G1期细胞比率.加温与DDP在诱导skov3细胞凋亡上表现为相加作用(0.85≤q≤1.15).41～42℃、30～90分钟加温与2.5 μg/ml DDP可协同诱导SK-OV-3细胞凋亡(q>1.15).结论热诱导肿瘤细胞凋亡并将细胞阻滞于G0/G1期是热疗杀伤卵巢癌细胞的重要机制.加温与DDP在诱导卵巢癌细胞凋亡和抑制细胞增殖等作用上具有相加或协同效应.     

靶向EGFR的干扰RNA对卵巢癌耐药细胞凋亡的影响

目的：探讨RNA干扰表皮生长因子受体（epidermal growth factor receptor,EGFR）的表达对多药耐药卵巢癌细胞株SKOV3/DDP凋亡的影响。方法：构建携带EGFR小发夹干扰RNA(small hairpin RNA, shRNA)的重组表达载体（pEGFRshRNA）,脂质体法转染入SKOV3/DDP细胞,同时设未转染对照组和非特异性干扰质粒CtrlshRNA对照组。RTPCR和免疫细胞化学法检测转染后SKOV3/DDP细胞内EGFR mRNA和蛋白的表达;流式细胞仪分析EGFR沉默后SKOV3/DDP细胞周期和凋亡率的变化。结果：与转染对照质粒组相比,转染pEGFRshRNA组细胞EGFR mRNA和蛋白的表达明显受抑制(P<001)。流式细胞仪检测结果显示：顺铂作用24 h后,pEGFRshRNA转染组细胞周期分布发生明显改变,与对照组和CtrlsiRNA组相比,G0/G1期细胞比例和凋亡率明显增加（P<0.01）,而S期细胞比例显著降低（P<0.01);凋亡率显著升高。结论：靶向EGFR的干扰RNA可抑制SKOV3/DDP细胞中EGFR的表达,调节耐药细胞周期,促进耐药细胞凋亡。     

WP1130通过下调β-catenin增强鼻咽癌细胞化疗敏感性

摘 要：［目的］探讨WP1130与顺铂联合用药对鼻咽癌细胞增殖和转移的影响及其可能机制。［方法］ CCK-8法检测鼻咽癌顺铂敏感细胞系HNE1和顺铂耐药细胞系HNE1/DDP的顺铂药物半抑制浓度（IC50）,以及不同浓度WP1130对鼻咽癌细胞活力的影响;选取适当药物浓度处理细胞,两种鼻咽癌细胞系分别分为4组：空白对照组（DMSO）、DDP单药组（6 ?滋mol/L顺铂）、WP1130单药组（1.25 ?滋mol/L WP1130）以及DDP+WP1130联合用药组（6 ?滋mol/L顺铂+1.25 ?滋mol/L WP1130）。MTT法检测鼻咽癌细胞增殖能力;细胞划痕实验检测鼻咽癌细胞迁移能力;Transwell侵袭实验检测鼻咽癌细胞侵袭能力;RT-PCR和Western blot实验检测顺铂耐药HNE1/DDP与亲本HNE1鼻咽癌细胞中β-catenin的表达情况以及不同药物处理对β-catenin的表达的影响。［结果］ 耐药HNE1/DDP细胞比亲本HNE1细胞对顺铂的耐受性提高了近5.7倍（5.17±0.33 vs 29.44±2.62,P＜0.01）。β-catenin在HNE1/DDP细胞中的mRNA表达上调约3.4倍（1.00±0.26 vs 3.44±0.39,P＜0.01）。与空白对照组和单药组相比,顺铂+WP1130联合用药组细胞中β-catenin的表达显著下调（HNE1：1.00±0.06 vs 0.78±0.07 vs 0.32±0.05,P＜0.05;HNE1/DDP：1.01±0.05 vs 0.57±0.07 vs 0.39±0.07,P＜0.05）,顺铂+WP1130联合用药组细胞增殖、迁移和侵袭能力均显著下降（P均＜0.01）。［结论］ WP1130与顺铂联合用药可以通过下调β-catenin的表达增强顺铂对细胞增殖、迁移和侵袭的抑制作用,从而增强鼻咽癌细胞的化疗敏感性。     

人参单体Rh2诱导人肺腺癌A549/DDP细胞凋亡的体外研究

背景与目的肺癌已成为人类癌症主要死亡原因之一.从植物中开发抗肿瘤药物是国内外抗肿瘤新药开发的一个热点.本研究的目的是观察人参单体Rh2诱导人肺腺癌A549/DDP细胞系凋亡的作用,并探讨其可能的分子机制.方法体外培养的人肺腺癌A549/DDP细胞经不同浓度梯度的人参单体Rh2干预,分别应用MTT实验观察其对细胞增殖的影响,用流式细胞术检测细胞周期、凋亡指数及相关基因表达的改变,以及用双抗体夹心ELISA法测定细胞上清液sApo-1/Fas浓度变化.结果①人参单体Rh2可抑制A549/DDP细胞的生长,并呈剂量、时间依赖作用.②人参单体Rh2处理A549/DDP细胞24 h,实验组凋亡指数显著高于对照组(P＜0.001),G0/G1期细胞比例显著高于对照组(P＜0.01),S期细胞比例显著低于对照组(P＜0.01),G2/M期细胞比例与对照组比较无显著性差异(P＞0.05).③实验组p53、Fas阳性表达率显著高于对照组(P＜0.01,P＜0.001),Bcl-2阳性表达率显著低于对照组(P＜0.001).④实验组A549/DDP细胞培养上清液于不同时间sApo-1/Fas含量均显著低于对照组(P＜0.05).结论人参单体Rh2具有诱导人肺腺癌A549/DDP细胞凋亡的作用,其分子机制可能是上调p53和Fas及下调Bcl-2的表达、通过Fas/FasL系统途径诱导细胞凋亡.     

人鼻咽癌多药耐药细胞CNE2/DDP与亲本细胞CNE2的生物学特性差异研究

目的:观察人鼻咽癌多药耐药细胞CNE2/DDP与亲本细胞CNE2的生物学特性差异,分析耐药细胞的特点。方法:比较两者在光镜下的形态、生长特点及体外克隆形成率的差异;流式细胞术分析两者细胞周期分布的变化、多药耐药蛋白P-170及ABCG2表达差异;采用MTT法分析耐药谱;动物实验观察两者成瘤差异。结果:光镜下CNE2/DDP细胞呈圆形,CNE2细胞呈多边形;CNE2/DDP生长相对缓慢;CNE2/DDP、CNE2细胞群体倍增时间分别为29.46 h、21.03 h;CNE2/DDP、CNE2细胞G0/G1期分别为(65.77±0.81)%、(44.9±2.21)%,S期分别为(23.63±0.42)%、(39.67±1.27)%,G2/M分别为(10.93±0.25)%、(13.23±0.31)%,两者相比差异均有显著性统计学意义(P<0.01)。CNE2/DDP、CNE2细胞的P-170和ABCG2表达率分别为(75.93±0.86)%、(2.83±0.40)%和(43.37±0.42)%、(4.07±0.59)%,两者差异有显著性统计学意义(P<0.01)。MTT法耐药谱分析表明,CNE2/DDP细胞成瘤前、后对DDP及5氟-尿嘧啶长春新碱(VCR)的耐药指数差异均无统计学意义。动物实验表明,1×106个CNE2/DDP、CNE2细胞皮下接种BALB/c裸鼠均成瘤,CNE2组肿瘤生长迅速。结论:CNE2/DDP细胞株具有多药耐药的特点,可用于耐药鼻咽癌的研究。     

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.     

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.     

Renal irradiation and the pharmacology and toxicity of methotrexate and cisplatinum

We used a rat model to study the effects of renal irradiation on the pharmacology of methotrexate (MTX) and cisplatinum (cis-Pt). Unanesthetized rats were given bilateral kidney irradiation (20 Gy in 9 fractions). At 9 months after irradiation, 3% of the animals had died and survivors showed moderately impaired renal function. At 15 months, 30% of the animals had died and survivors showed severely impaired renal function. Some animals were given i.v. MTX 1 week to 15 months after irradiation. In irradiated rats, the area under the MTX plasma clearance curve equaled that of controls through 6 months, and was significantly above controls from 9 months on. Other animals were given i.p. cis-Pt 1 week to 9 months after irradiation. The acute toxicity of cis-Pt was the same in control and irradiated rats when cis-Pt was given immediately before or after irradiation. Beginning 3 months after irradiation there was a progressive increase in cis-Pt toxicity and a simultaneous decrease in urinary platinum excretion. Irradiated animals that survived cis-Pt treatment showed increased radiation nephritis; the greatest effect occurred when cis-Pt was given 3 months or more after irradiation. MTX and cis-Pt clearance decreased when renal dysfunction was first observed and changes in renal function preceded changes in drug clearance and toxicity.     

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。     

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.