Nuclear cyclin D1 overexpression is a critical event associated with cell proliferation and invasive growth in gallbladder carcinogenesis

Cyclin D1 overexpression is remarkably frequent in several human carcinomas and is believed to be a critical event in oncogenesis. We examined cyclin D1 expression, p53 expression, and the Ki-67 labeling index by immunostaining in human gallbladder mucosa in conditions varying from normal to malignant tissue. We also examined K-ras codon 12 mutations in these tissues with a two-step polymerase chain reaction. Nuclear cyclin D1 overexpression was observed in 48% of carcinomas occurring independently of adenoma, but not in adenomas, carcinomas arising in adenomas, or nonneoplastic lesions. Cytoplasmic cyclin D1 overexpression was observed in about 15% of abnormal specimens, irrespective of the type of epithelial abnormality. Carcinomas showing nuclear cyclin D1 overexpression had significantly higher Ki-67 labeling indexes than those with no overexpression. Moderately to poorly differentiated adenocarcinomas showed a higher incidence of nuclear cyclin D1 overexpression than papillary to well differentiated carcinomas. Specimens with cyclin D1 overexpression showed a high incidence of lymph permeation, venous permeation, and lymph node metastasis. We conclude that nuclear cyclin D1 overexpression is a critical event importantly associated with cell proliferation and invasive growth in gallbladder carcinogenesis, and that cyclin D1 immunostaining may become a useful marker for evaluating gallbladder carcinomas. Received: March 9, 1999 / Accepted: July 23, 1999     

Efficacy of β-diketonato complexes of titanium,zirconium, and hafnium against chemically induced autochthonous colonic tumors in rats

Summary Bis--diketonato complexes of titanium, zirconium, and hafnium were tested against autochthonous colorectal tumors in rats. The model was found to reflect the clinical situation most closely. of the compounds tested, budotitane was the most effective in terms of decrease in tumor weight and number and in increasing the lifespan of the treated animals. The therapeutic efficiency was superior to that of 5-fluorouracil, which so far has been the drug with the best activity in patients suffering from colon cancer.Abbreviations Ti(bzac)₂Cl₂ dichlorobis(1-phenylbutane-1,3-dionato)-titanium(IV) - Ti(bzac)₂(OEt)₂, budotitane, (INN) diethoxybis(1-phenylbutane-1,3-dionato)-titanium(IV) - Zr(bzac)₂Cl₂ dichlorobis(1-phenulbutane-1,3-dionato)-zirconium(IV) - Hf(bzac)₂Cl₂ dichlorobis(1-phenylbutane-1,3-dionato)-hafnium(IV) - AMMN acetoxymethylmethylnitrosamine; 5-FU - 5-FU 5-Fluorouracil - DFUR 5-deoxy-5-fluorouridine - CPA cyclophosphamide - cis-DDP, Cisplatin cis-diamminedichloroplatinum(II) - INN international nonproprietary name Dedicated to Professor Norbert Brock on the occasion of his 75th birthday     

Saccharomyces boulardii in Maintenance Treatment of Crohn’s Disease

The possible role of Saccharomyces boulardii, a nonpathogenic yeast with beneficial effects on the human intestine, in the maintenance treatment of Crohns disease has been evaluated. Thirty-two patients with Crohns disease in clinical remission (CDAI < 150) were randomly treated for six months with either mesalamine 1 g three times a day or mesalamine 1 g two times a day plus a preparation of Saccharomyces boulardii 1 g daily. Clinical relapses as assessed by CDAI values were observed in 37.5% of patients receiving mesalamine alone and in 6.25% of patients in the group treated with mesalamine plus the probiotic agent. Our results suggest that Saccharomyces boulardii may represent a useful tool in the maintenance treatment of Crohns disease. However, in view of the products cost, further controlled studies are needed to confirm these preliminary data.     

Interphase FISH to detect PBX1/E2A fusion resulting from the der(19)t(1;19)(q23;p13.3) or t(1;19)(q23;p13.3) in paediatric patients with acute lymphoblastic leukaemia

Approximately 6% of paediatric patients with precursor B-cell acute lymphoblastic leukaemia (B-ALL) harbour a rearrangement involving the gene regions of PBX1 (1q23) and E2A (19p13.3) which is visualized cytogenetically either as a der(19)t(1;19)(q23;p13.3) or the less common balanced t(1;19)(q23;p13.3). Unfortunately, no commercial dual-colour, double fusion fluorescence in situ hybridization (D-FISH) strategies are available to detect this recurrent anomaly. Therefore, we have created a D-FISH assay to detect these translocations and monitor minimal residual disease. This probe set was created using four bacterial artificial chromosomes (BACs) corresponding to the PBX1 gene region at 1q23 and four BACs corresponding to the E2A gene region at 19p13.3. We analysed 30 negative bone marrow controls and 20 diagnostic and post-treatment specimens from 13 paediatric B-ALL patients with a cytogenetically defined 1;19 translocation. Once unblinded, the results demonstrated that our D-FISH method effectively identified all diagnostic samples as abnormal and identified disease in four post-treatment samples that were previously considered to be normal by conventional cytogenetic analysis. The development of this FISH strategy for the detection of der(19)t(1;19)(q23;p13.3) and t(1;19)(q23;p13.3) proved to be an effective technique, allowing both the detection of disease in diagnostic samples and in post-treatment samples.     

尼古丁对口腔癌前病变细胞中Prx1表达及MAPK信号通路活化的影响

目的 通过检测尼古丁对口腔癌前变细胞株DOK细胞中抗氧化蛋白Prx1表达及MAPK三种激酶JNK、ERK、p38的磷酸化水平的影响,探讨尼古丁对口腔癌前病变细胞凋亡的作用机制,以期为口腔癌前病变的防治提供科学依据及新的生物靶点.方法 1μmol/L尼古丁长期处理口腔癌前病变细胞DOK 7天,采用荧光定量PCR和Western Blot的方法,检测尼古丁组与对照组中Prx1 mRNA与蛋白的表达及磷酸化JNK、ERK、p38蛋白(p-JNK、p-ERK、p-p38)的表达情况.结果 倒置显微镜下观察,1μmol/L尼古丁长期处理口腔癌前变细胞株DOK细胞7天后,细胞形态未见明显变化.与对照组相比,尼古丁组DOK细胞中Prx1 mRNA及蛋白表达水平均显著增高(P=0.011,P=0.044).p-p38(P=0.030)和p-JNK(P =0.017)蛋白表达与对照组相比降低,p-ERK的蛋白表达增高(P=0.035),差异有统计学意义.结论 Prx1、JNK、ERK及p38可能在烟草相关口腔癌前病变细胞凋亡中发挥重要作用.     

靶向p21基因saRNA抑制肝癌细胞生长实验研究

目的 探讨RNA激活p21对肝癌HepG2、Hep3b和SMMC-7721细胞生长和侵袭力的影响.方法 化学合成靶向p21的saRNA、阴性对照dsRNA,将其转染肝癌细胞系HepG2、Hep3b和SMMC-7721.每个细胞系分为3组,分别为p21-322组、阴性对照组和空白对照组,每组复3孔;p21-322组和阴性对照组分别采用p21-322 saRNA和阴性对照dsRNA进行转染,空白对照组不干预.转染后72 h,采用RT-PCR和Western blot检测各组细胞中的p21 mRNA和P21蛋白表达水平,采用MTT法检测细胞生长情况及划痕实验观察细胞侵袭能力的变化.结果 HepG2、Hep3b和SMMC-7721细胞p21-322组p21 mRNA相对表达水平分别为23.43±2.29、16.87±1.61、31.77±5.06,P21蛋白相对表达水平分别为55.93±12.66、32.91±5.17、24.96±6.81;空白对照组分别为3.53±0.07、2.39±0.02、5.70±0.89,3.21±0.03、2.91±0.14、4.15±0.12;阴性对照组分别为3.87±0.97、2.57±0.71、5.87±1.73,3.11±0.70、3.01±0.97、5.13±2.14;p21-322组p21 mRNA和蛋白相对表达水平明显高于空白对照组和阴性对照组(P＜0.01),空白对照组与阴性对照组比较差异无统计学意义(P＞0.05);细胞转染后第6天时HepG2、Hep3b、SMMC-7721细胞p21-322转染组细胞平均生长抑制率分别为41％、48％和52％;HepG2细胞p21-322组24 h后空白区域占原划痕区域面积的百分比((76±11)％)明显高于空白对照组((13±6)％)和阴性对照组((17±8)％)(P＜0.01),阴性对照组与空白对照组比较差异无统计学意义(P＞0.05).结论 靶向p21的RNAa能抑制肝癌细胞的生长和侵袭力,p21可作为一个具有肝癌治疗应用价值的靶基因.