Screening and identification differentially displayed genes in fore stomach carcinoma of mice

Objective: To screen and identify key genes differentially displayed in mouse fore stomach carcinoma, in order to elucidate the molecular mechanism underlying carcinogenesis.Methods: The animal models complied with each period of NIH mouse fore stomach carcinoma induced by N-Nitrososarcosineethylester (NSEE) were used in this study. The mice were euthanized on days 14, 28, 56, 77 and 84, respectively, after NSEE-piped treatment, and classified according to their pathologies. The differentially expressed genes were isolated from both normal and morbid tissues by mRNA differential display technique and screened by using Reverse Northern blot. Bioinformatics were employed to analyze the results observed. After identification, ten fragments were cloned and matched with GENEBANK database through homologous analysis.Results: One gene was found identical to splicing factor 3b subunit 1 (Sf3b1), while seven fragments hold the homology of known cDNA clones. In contrast, other two fragments had extremely low identity to any genes registered in GENBANK databases.Conclusion: It is the first time to demonstrate in this study that splicing factor3b, subunit1 (Sf3b1) is related to mouse fore stomach carcinoma. Furthermore, ESC-3 and ESC-4 are suggested to contribute to the development of mouse fore stomach carcinoma, thus may be candidates of new targets of oncogenes.     

左金丸对小鼠S180肿瘤细胞获得性多药耐药P—gp的表达及血清肿瘤标志物的影响

目的：研究左金丸、黄连和吴茱萸对小鼠获得性多药耐药S180腹水肿瘤细胞生长的抑制作用,以及对肿瘤细胞P—gP的表达和血清4种肿瘤标志物：（TM）酸性磷酸酶（ACP）、碱性磷酸酶（AKP）、醛缩酶（ALD）和乳酸脱氢酶（LDH）活力的影响。方法：PFC方案建立S180多药耐药细胞模型,将耐药后S180细胞悬液以每只小鼠0．2ml接种于腹腔,黄连、吴茱萸及左金丸灌胃给药7d,观察小鼠体重、脾指数、肝指数、生命延长率（ILS）,流式细胞仪测定P-糖蛋白（P—gp）的表达率,试剂盒检测血清中肿瘤标志物的活力。结果：左金丸给药组对小鼠S180多药耐药细胞增长有明显的抑制作用,能有效降低肝指数,提高脾指数,提高生命延长率（ILS）（60．26％）的作用明高于黄连（21．85％）和吴茱萸单独给药（20．53％）（P〈0．01）。同时,左金丸给药组能有效逆转$180肿瘤细胞中P—gp的表达（2．69％）至接近S180细胞对照组水平（1．65％）,显著低于模型组（16．07％）,也强于黄连（5．69％）及吴茱萸（9．15％）单独给药时的逆转效果。并且左金丸可有效降低血清中ALD（75．944-12．47U·L^-1）和LDH（1632．924-494．48u·L^-1）的活力,提高血清中ACP（103．284-10．36U·100ml^-1）和AKP（34．564-4．91U·100ml^-1）的活力,与黄连、吴茱萸给药组比较有显著性差异（P〈0．01）。结论：黄连和吴茱萸合用能产生明显的配伍协同抗肿瘤作用,对P—gp的表达和血清中肿瘤标志物的影响可能是其抗肿瘤和逆转多药耐药的潜在机制。     

微囊藻毒素LR对人FL和HL7702细胞PP2A各亚基转录水平的影响

背景与目的： 研究微囊藻毒素LR(microcystin LR,MCLR)对正常人羊膜细胞FL和肝实质细胞HL7702蛋白磷酸酶2A(PP2A)各亚基mRNA表达的影响。材料与方法： FL和HL7702经100 nmol/L MCLR作用24 h后,采用实时定量PCR方法检测PP2A各亚基mRNA表达的变化。实验并设溶剂对照组。 结果： FL细胞PP2A各亚基mRNA表达部分上升,部分下降;而HL7702细胞PP2A各亚基mRNA表达均上升。 结论： MCLR对肝来源细胞的特异性作用可能与其肝脏毒性有关。     

复合核黄素抗亚硝胺致癌作用的动物实验研究

本文应用ＮＳＥＥ诱发ＮＩＨ小鼠前胃鳞癌的动物模型，观察了复合核黄素和纯核黄素对小鼠前胃鳞状上皮癌变的影响。结果表明，复合核黄素对ＮＳＥＥ诱发小鼠前胃鳞癌有明显抑制作用，其抑制率为５４．８４％（Ｐ＜０．０５）。而纯核黄素未发现明显的阻断癌变的作用。提示，其有效成分可能不是核黄素，而是核黄素以外的其它成分或是核黄素与其它成分的协同作用。本研究为食管癌的预防研究提供了实验依据。     

大肠腺瘤及其癌变的临床病理分析

目的探讨大肠腺瘤及腺瘤癌变与临床病理特征的关系。方法回顾性分析大肠腺瘤及癌变病例与腺瘤大小、解剖部位、外形、病理类型的关系。结果776例大肠腺瘤患者发生癌变者为42例,癌变率为5.4%,绒毛状腺瘤癌变率最高,管状腺瘤癌变率较低;直肠腺瘤癌变率均高于右半结肠(P<0.05);大肠腺瘤癌变率随着腺瘤的增大而升高,并且广基形腺瘤比长蒂形腺瘤更易癌变。大肠腺瘤的癌变率随异型增生程度的增高而增高(P<0.05)。结论大肠腺瘤癌变与腺瘤大小、解剖部位、外形、病理类型及腺瘤异型增生程度等因素有关。     

膀胱癌变过程中PCNA表达的研究

为了解膀胱癌变过程中细胞增殖状态及其与肿瘤生物学行为、复发的关系，作者采用免疫组化方法检测了正常上皮、增生上皮、乳头状瘤和癌组织中的增殖细胞核抗原表达。增殖细胞核抗原指数在四种组织中呈增高趋势，并且与肿瘤病理分级、分期密切相关，但与肿瘤大小、数目无相关。复发者的增殖细胞核抗原指数显著高于无复发者。结果提示，膀胱癌变过程中细胞增殖能力不断增强，增殖细胞核抗原指数可以作为肿瘤生物学行为、判断肿瘤复发的一项有价值的标记物。     

Recovery of hyperplastic responsiveness in rat liver after dosing with the peroxisome proliferator methylclofenapate

Methylclofenapate (MCP) was administered daily by gavage (25 mg/kg) for 7 days to groups of adult male rats. Dosing was interrupted for 28, 35, 56, 70 or 84 days and then resumed (25 mg/kg by gavage at 0 and 24 h). During the second period of dosing animals were killed in groups of three at 6, 12, 18, 24, 30, 36, 42 and 48 h after the resumption of dosing. Hepatocytes in S-phase, labelled with bromodeoxy-uridine, were analysed by flow cytometry, cell sorting and microscopy. It was observed that total S-phase activity was just significantly elevated (approximately 20% of maximum) over corn oil controls after an interval of 28 days between initial and subsequent dosing periods. After an interval of 35 days total S-phase activity was approximately 65% of maximum, and full hyperplastic responsiveness, equal to that observed in naive animals given MCP, was detected after interruptions in dosing of 56, 70 and 84 days. The recovery of S-phase responsiveness during the interruptions in dosing was accompanied by an increase in the proportion of 2 X 2N hepatocytes from approximately 10% in animals dosed continuously with MCP, to approximately 11.4% after 28 days interruption, 17% after 35 days and control levels (approximately 20%) after 56, 70 and 84 days. Irrespective of the magnitude of the hyperplasia elicited by the second period of dosing with MCP, the proportion of 2 X 2N cells was reduced to the same levels as those observed in animals dosed continuously with MCP (approximately 10%). Very low S-phase activity (0.05%) was observed in animals dosed continuously with MCP, this level of activity being similar to that in animals given corn oil continuously.     

The stimulating effect of acetic acid, alcohol and thermal burn injury on esophagus and forestomach carcinogenesis induced by N-nitrososarcosin ethyl ester in rats

Five groups of outbred white male rats were given N-nitrososarcosin ethyl ester (NSEE) i.g. for 4 or 6 months at a daily dose of 50 mg/kg of body wt. 5 days/week. Some groups of animals were given a 3% water solution of acetic acid or a 40% solution of ethanol i.g. for 8 months from the beginning of the experiment. The remaining groups of these rats received controlled local thermal burn injury of the esophageal mucosa 15 days before the beginning of the experiment. Acetic acid solution increased the multiplicity of benign and malignant tumors as well as carcinoma incidence in the esophagus. Ethanol in combination with NSEE did not influence carcinogenesis in the esophagus but increased the incidence of leukokeratosis and the multiplicity of forestomach papillomas. In rats treated with NSEE after thermal burn injury, a significant increase in the frequency and multiplicity of papillomas was found in the burn zone.     

Treatment of recurrent gliomas with eflornithine.

BACKGROUND: Oral eflornithine in combination with intravenous mitoguazone (methylbisguanylhydrazone) has shown activity against recurrent anaplastic gliomas. Eflornithine alone, however, has not been evaluated against recurrent gliomas. PURPOSE: This study compared the antitumor activity of oral eflornithine with that of oral eflornithine combined with intravenous mitoguazone in the treatment of patients with recurrent or progressive glioblastoma multiforme as well as nonglioblastoma anaplastic gliomas. METHODS: During the 1st year of therapy with eflornithine alone, the drug was given at a dose of 3.6 g/m2 on days 1-14, 22-35, and 43-56 every 8 hours; cycles were repeated every 63 days until progression. For the 2nd year, the drug was given on days 1-14, 29-42, and 57-70, with 84 days between cycles. For the 1st and 2nd years of eflornithine-mitoguazone therapy, eflornithine was given at 1.8 g/m2 on the same schedule. Mitoguazone was given intravenously at 200 mg/m2 on the final day of each 2-week sequence of eflornithine therapy. Response was determined by evaluating changes in the size of contrast-enhanced neuroimages. RESULTS: Because of two cases of lethal hepatic necrosis, the initial random allocation of patients to the eflornithine-mitoguazone arm was stopped after 23 patients had been accrued. Ninety-eight patients were entered in the eflornithine arm; 80 patients (36 glioblastoma multiforme patients and 44 anaplastic glioma patients) were assessable for response. Antitumor activity (partial response, minor response, and stable disease) was seen in 45% of the patients with anaplastic gliomas, for a median of 49 weeks, but in only 17% of patients with glioblastoma multiforme (median not attained). Twenty-one (20%) of the patients with anaplastic glioma and 33% of the patients with glioblastoma multiforme were removed from the study before completing the first 8-week course of therapy because of neurological deterioration and tumor progression by the 5th week of treatment. CONCLUSION: This study suggests that eflornithine alone is an effective palliative therapy for recurrent anaplastic gliomas. Additional studies are needed to confirm our finding.     

HSP70、c-fos、PCNA 在乳腺癌中的表达研究

 目的　研究 HSP70、c- fos、PCNA在乳腺癌组织中表达的相互关系。方法　应用免疫组化 S- P法检测 6 6例乳腺癌标本中 HSP70、c- fos、PCNA的表达情况。结果　在乳腺癌组织中 ,HSP70表达阳性率为 71 % (4 7/ 6 6 ) ;c- fos阳性率为 5 6 % (37/ 6 6 ) ;PCNA阳性率为 6 5 % (4 3/ 6 6 )。经统计学分析 ,HSP70阳性等级与 c- fos阳性等级具有显著相关性 (P<0 .0 1 ) ;HSP70阳性等级与PCNA阳性等级也具有显著相关性 (P<0 .0 1 )。结论　在乳腺癌组织中 ,HSP70的表达与 c- fos的表达存在着一定的相关性 ,HSP70与乳腺癌细胞增殖可能相关。     

大肠腺癌组织中VASP和PCNA的表达

目的研究大肠腺癌组织中VASP和PCNA的表达及临床意义。方法采用SP免疫组化染色法检测25例大肠腺癌组织中VASP及PCNA的表达。结果VASP阳性21例(84%),PCNA阳性18例(72%),两者均为阳性16例(64%),均为阴性2例(8%);VASP表达率和PCNA表达率之间有明显正相关性(P<0.05);VASP表达与PCNA表达均与大肠腺癌分化程度显著相关(P<0.05),在合并后的Duke's分期A、B和C、D期间有显著性差异(P<0.05)。结论VASP随着大肠腺癌分期、分级增加而表达增强的特性提示,VASP变化可能成为辅助判断大肠腺癌的转移和增殖特性的一个新指标。     

生后小鼠肾脏皮质发育过程中的细胞增殖

目的 :观察小鼠出生后肾脏皮质发育过程中增殖细胞核抗原 (proliferatingcellnuclearantigen ,PCNA)的表达特征 ,探讨出生后小鼠肾脏皮质发育过程中细胞增殖的规律。 方法 :应用免疫组织化学技术检测小鼠出生后 1、 3、 7、 14、 2 1、 2 8和 70天肾脏皮质PCNA的表达。结果 :小鼠出生后 1～ 70天 ,皮质中的生肾区、肾小体、肾小管及髓放线中PCNA阳性细胞的表达具有一定的规律 ,早期PCNA阳性表达丰富 ,随着肾脏发育逐渐成熟而表达减弱直至消失 ,在成年鼠肾脏皮质中 ,没有检测到PCNA阳性细胞。结论 :出生后小鼠肾脏皮质中的生肾区、肾小体、肾小管及髓放线的细胞增殖规律是由高逐渐降低的 ,直至成年完全停止。     

非小细胞肺癌后程加速超分割放疗疗效观察

目的：通过前瞻性的研究非小细胞肺癌（ＮＳＣＬＣ）后程加速超分割放疗的疗效，寻找ＮＳＣＬＣ的最佳治疗手段。方法：对１９９３年１月～１９９６年７月我院收治的７０例ＮＳＣＬＣ患随机分组研究。常规组３５例：用＾６０Ｃｏ或８Ｍｖ－ｘ，５次／Ｗ，２Ｇｙ／次，总剂量７０Ｇｙ／３５次／７～８Ｗ。后程加速超分割组３５例：放疗前程为常规分割，２Ｇｙ／次，４０Ｇｙ／２０次／４Ｗ后改野，５天／Ｗ，２次／天，１．８Ｇｙ／     

细胞增殖与凋亡在小鼠生后肾脏髓质发育过程中的意义

观察小鼠出生后肾脏髓质发育过程中增殖细胞核抗原(proliferatingcellnuclearantigen,PCNA)的表达及细胞凋亡的特征,探讨出生后小鼠肾脏髓质发育过程中细胞增殖与凋亡的规律及其关系。应用免疫组织化学技术和原位末端标记法(TUNEL法)分别检测小鼠出生后1、3、7、14、21、28和70天肾脏髓质中的增殖细胞及凋亡细胞。结果显示,(1)小鼠出生后肾脏髓质发育中PCNA阳性细胞的表达具有一定的规律,早期PCNA阳性表达丰富,随着肾脏发育逐渐成熟而表达减弱;(2)在细胞增殖的同时也存在着细胞凋亡现象,细胞凋亡指数于生后第7天达到高峰,以后表达逐渐减弱。细胞增殖与凋亡在小鼠生后肾脏髓质发育的整个过程中普遍存在,早期细胞增殖旺盛,增殖高峰之后出现凋亡高峰,晚期两者活动均减弱。     

Carcinogenic and promoting effects of Roussin red methyl ester (RRME) on the forestomach epithelium of mice and esophageal epithelium of rats, and its inhibition by retinamide and vitamin C

Carcinogenic and promoting effects of RRME as isolated from the pickled vegetables in Linxian County, a high incidence area of esophageal cancer, were studied in mice and rats. RRME alone did not cause tumor in the forestomach of mice and esophagus of rats. When the mice were intubated with a single dose of nitroso-sarcosine-ethylester (NSEE), the incidence of the forestomach carcinoma was only 9.5%. However, when the mice were given gastric doses of RRME after one single dose of NSEE, the incidence was increased to 41.0%. In rats, the tumor incidence was 5.3% in nitroso-methylbenzylamine (NMBzA) group, while in NMBzA kRME group, it was 20.7%. In rats intubated with NSEE for 7 times, no carcinoma appeared in esophageal epithelium; while followed by gastric doses of RRME, the incidence of esophagus carcinoma increased up to 63.2%. The experimental results show that RRME has distinct promoting effect on the process of cocarcinogenesis initiated by NSEE and NMBzA in the forestomach of mice and esophagus of rats, but without carcinogenic effect itself. Retinamide (RI) and massive dose of vitamin C showed an obviously inhibitory effect on promoting action of RRME in rats.(ABSTRACT TRUNCATED AT 250 WORDS)     

Carcinogenic and promoting effects of fish juice, preserved rice and salted dry fish on the forestomach epithelium of mice and esophageal epithelium of rats

The carcinogenic and promoting effects of fish juice, preserved rice and salted dry fish from Nanau county, Guangdong province, a high incidence area of esophageal cancer, were studied in mice and rats. The homemade fish juice as well as fish juice in market, whether or not added with NaNO2, did not cause tumor in the forestomach of mice and the esophagus of rats. When the mice were intubated with an initiator, nitrososarcosinethylester (NSEE) twice, no carcinoma was found at the end of the experiment (D 120). Only papilloma appeared in the forestomach epithelium. The incidence was only 37.5%. However, when the mice were intubated with NSEE for 2 times followed by gastric doses of homemade fish juice, the tumor incidence in the forestomach was increased to 89.7%, in which 20.5% was carcinoma. The tumor and carcinoma incidences of initiator (NSEE and NMBzA) group and initiator + market fish juice group in mice and rats were without significant difference. The experimental results show that the homemade fish juice proved distinct promoting effect on the process of cocarcinogenesis initiated by NSEE in the forestomach of mice, while the market fish juice has no significant promoting effect on the forestomach epithelium of mice and the esophageal epithelium of rats. NSEE induced 31.6% carcinoma in the forestomach epithelium of mice on standard diet. While in mice fed with preserved rice and salted dry fish, the carcinoma incidence was increased to 63.6%. It appears that preserved rice and salted dry fish have promoting effect on the process of cocarcinogenesis initiated by NSEE in the forestomach of mice.(ABSTRACT TRUNCATED AT 250 WORDS)     

热休克诱导人肝癌HepG2细胞HSP70和P-gp的表达及槲皮素对二者的抑制作用

背景与目的:研究提示热休克蛋白70(heatshockprotein70,HSP70)可抑制细胞凋亡,降低肿瘤热疗、化疗的疗效,但其与细胞化疗耐受性的内在关系尚不甚明确;P-糖蛋白(P-glycoprotein,P-gp)是一种耐药蛋白,在肿瘤耐药中起重要作用。本研究拟通过热休克诱导人肝癌HepG2细胞表达HSP70和P-gp,并观察槲皮素(quercetin,Qu)对二者表达的抑制效应。方法:恒温水浴法,42℃热休克HepG2细胞90min,分别采用免疫组化、流式细胞术检测热休克前及热休克后2h、4h、8h、12h、24h时HSP70和P-gp的表达情况;并于热休克前1h应用不同浓度槲皮素作用于HepG2细胞,观察其热休克后4h二者的表达。结果:(1)热休克诱导后,免疫组化显示细胞内HSP70表达增多,“核内迁移”;流式细胞术示HSP70阳性细胞数升高,4h达到最高(11.47%),较对照组(6.64%)升高近一倍(P<0.01),24h后回落到低值;P-gp阳性细胞数随HSP70升高后升高,12h达到最高值(96.31%),较对照组(33.95%)升高近两倍(P<0.01),24h后回落到低值。(2)热休克前应用槲皮素能有效抑制热休克诱导HSP70和P-gp的过量表达,以100～200μmol/L槲皮素作用明显(P<0.01),呈浓度依赖性。结论:热休克可诱导HepG2细胞HSP70和P-gp的过量表达,P-gp的表达可能与HSP70的表达有关;槲皮素可阻断热休克诱导HepG2细胞HSP7     

增殖细胞核抗原在卵巢上皮性癌的表达及其临床意义

探讨增殖细胞核抗原表达与卵巢上皮性癌生物学行为及预后的关系。材料与方法：采用ＬＳＡＢ免疫组化方法检测８４例石蜡包埋的卵巢上皮性癌组织中增殖细胞核抗原的表达,分析其与患者的年龄、临床分期、组织分化程度、组织学类型及预后之间的关系。     

Secretory and mitotic response of the male rat pituitary gland to repeated doses of oestrogen

Pituitary weight, pituitary mitotic index and pituitary and serum concentrations of growth hormone and prolactin were measured at 14-day intervals in male rats given 10 mg diethylstilboestrol dipropionate SC every 14 days for a total period of 70 days. Mean pituitary weight reached a maximum of 21 mg on day 42 and no pituitary gland became a tumour according to a criterion of weight greater than 30 mg. Mitotic index was greatly increased until day 70 when it diminished significantly. Pituitary concentration of growth hormone decreased to maintain a low constant level from day 28 onwards while pituitary prolactin concentration rose to a maximum on day 42. Serum growth hormone was elevated on day 14 and then gradually diminished, while serum prolactin continued to increase until day 42. Pituitary weight and serum prolactin were significantly correlated and pituitary mitotic index was negatively correlated with pituitary growth hormone concentration. The state of apparent equilibrium in mass reached by the pituitary glands from day 42 until day 70, despite greatly increased secretory and mitotic activity, suggests that further disturbance of pituitary function is necessary to bring about net gain in cell numbers and lead to tumour formation.     

人脑胶质瘤组织中nm23与PCNA的表达及其意义

背景与目的:脑胶质瘤极少发生颅外转移,死亡的主要原因是肿瘤的原位复发,因此,通过检测基因表达进一步了解其生物学特性很重要。本研究旨在探讨胶质瘤组织中肿瘤转移抑制基因(non-metastasis,nm23)、增殖细胞核抗原(proliferatingcellnuclearantigen,PCNA)的表达及其对肿瘤恶性程度的判断、患者预后评估和复发预测的意义。方法:用免疫组化SP法测定50例不同恶性程度的胶质瘤组织中nm23、PCNA的表达情况。结果:(1)低度恶性胶质瘤标记指数(labelindex,LI)nm23为3.40±0.27,PCNA为3.60±0.05;高度恶性胶质瘤nm23LI为1.72±0.18,PCNALI为6.20±0.23,有显著性差异(P<0.05);(2)25例低度恶性胶质瘤中nm23阳性14例(56%),PCNA阳性16例(64%);而25例高度恶性胶质瘤中nm23阳性3例(12%),PCNA阳性22例(88%)。低度和高度恶性胶质瘤两组nm23、PCNA表达阳性率有显著性差异(P<0.05);(3)复发组9例,无nm23阳性者(0%),PCNA全部阳性(100%),未复发组8例,4例nm23阳性(50%),4例PCNA阳性(50%),两组nm23、PCNA的表达阳性率均有显著性差异(P<0.05);(4)胶质瘤nm23与PCNA的标记指数呈负相关(r=-0.5335,P<0.001)。结论:(1)nm23的表达随胶质瘤恶性程度增加而下降;(2)PCNA的表达随胶质瘤恶性程度的增加而升高;(3)nm23、PCNA可作为胶质瘤恶