p53 Protein expression in laryngeal squamous cell carcinomas bearing wild-type and mutated p53 gene

We performed an immunohistochemical analysis to investigate the expression of p53 protein in a panel of 18 laryngeal squamous cell carcinomas, 15 primary tumours and three in relapse, previously analysed by us for the presence of p53 gene mutations. Dysplastic and/or normal surrounding mucosa was evaluated in 15 different tumours. The results of our study are the following: (1) expression of p53 protein was observed in one out of five tumours positive for p53 gene mutations (20%) and in 10 out of 13 (80%) negative cases; (2), p53 protein over-expression was frequently observed in normal and/or dysplastic mucosa surrounding either wild-type (7/11) or mutated p53 tumours (2/4); (3), p53 immunoreactive cells showed a pattern of distribution in normal and mildly/moderately dysplastic mucosa (basal layers), different from that in severely dysplastic mucosa (whole thickness). These data further support the hypothesis that p53 protein over-expression may be a marker of the earliest phases of multistep tumorigenesis in laryngeal squamous cell carcinoma.     

P16、Cyclin D1蛋白表达与胰腺癌生物学行为

目的 探讨P16、CyclinD1蛋白表达水平与胰腺癌发生、发展及预后的关系。方法 应用免疫组织化学SP法结合微波抗原修复法对53例原发性胰腺癌和42例胰腺良性病变及正常组织中P16蛋白及CyclinD1蛋白进行检测。结果 胰腺癌组织、胰腺良性病变和正常组织中P16蛋白的阳性率分别为66．04％和87．71％（P＜0．05）；CyclinD1表达的阳性率分别为52．83％和23．81％（P＜0．05），P16及CyclinD1蛋白表达与胰腺癌的组织分化、病理分期及术后生存期密切相关。结论 P16及CyclinD1蛋白在胰腺癌中的表达可判断胰腺癌的预后。     

毛喉萜对人肝癌细胞增殖影响的实验研究

目的　探讨毛喉萜 (Forskolin)的抗肝癌作用。方法　采用克隆形成、MTT比色法观察毛喉萜对SMMC 772 1细胞的作用 ,并利用免疫细胞化学方法检测其对rasp2 1、p5 3蛋白表达的影响。结果　毛喉萜可明显抑制SMMC 772 1细胞的增殖 ,其抑制率与剂量呈正相关 (P <0 .0 1 ) ,经毛喉萜处理后 ,rasp2 1、p5 3蛋白表达均有明显下降。结论　毛喉萜能明显抑制肝癌SMMC 772 1细胞增殖 ,其作用可能与降低rasp2 1、p5 3蛋白表达有关。     

动脉灌注p53基因治疗晚期肝癌的初步临床应用

目的 初步观察动脉灌注p53基因治疗晚期肝癌的疗效.方法 晚期肝癌30例,治疗组14例,对照组16例.根据造影表现决定灌注药物的靶动脉,治疗组在靶动脉注入p53基因,每次用1012VP,加羟基喜树碱20 mg,每周1次,连用3周为1疗程,14例患者分别接受1～4疗程的治疗.对照组给予羟基喜树碱20 mg肝动脉灌注.结果 治疗组生存期最小14 d,最长405 d,平均238.1 d.对照组生存期最小18 d,最长167 d,平均80.7 d.两者相比较,P＜0.05,有显著差异.结论 p53基因治疗晚期肝癌有效.     

电离辐射调控脂质体介导的p16基因在HeLa细胞中的表达

目的探讨不同剂量γ射线照射后诱导脂质体介导的p16基因在HeLa细胞中的表达及抗癌作用。方法用脂质体Lipofectamin介导重组质粒Egr-p16转染人HeLa细胞，采用RT-PCR的方法检测了。Coγ射线照射转染后的人HeLa细胞剂量效应和时程变化，用细胞计数检测细胞增殖的变化，用流式细胞术检测细胞周期的变化。结果研究证实0．5～8Gv照射后p16的转录水平高于对照，在2～4Gy照射后达到峰值，2Gy照射后2—24h高于对照组，在照射后4h达到峰值，然后逐渐下降。细胞生长曲线显示体外稳定转染联合。Coγ射线照射对HeLa细胞的增殖具有明显的抑制作用。细胞周期变化显示转染后的HeLa细胞经过照射后G0／G1期比例呈现剂量依赖性的下降，而S期则出现剂量依赖性的增加，出现明显的S期阻滞，G2／M期在2Gy出现明显的阻滞，在5和10Gy时逐渐下降，但是仍然高于对照组。结论^60Coγ射线可诱导转染的HeLa细胞p16转录水平的增强，同时在细胞增殖上出现明显的变化。     

p38丝裂原激活蛋白激酶对人胚胎肾293细胞一氧化氮合酶基因转录的调控

目的探讨p38 丝裂原激活蛋白激酶(MAPK)家族四种亚型对诱导型一氧化氮合酶(iNOS)基因的转录调控。方法以人胚胎肾293(HEK 293)细胞为靶细胞,采用脂质体(LPS)介导的细胞基因共转染技术、荧光素酶报告基因技术,分别将FLAG-tagged p38 MAPK 4种亚型、含有鼠iNOS基因启动子区的荧光素酶报告基因质粒(piNOS-Luc)、空载体(pcDNA3)、β-半乳糖苷酶表达质粒(pCMV-β)共转染,检测并比较荧光素酶相对活性。结果(1)未加刺激时,在HEK 293细胞中,p38 MAPK中仅有p38α能够诱导iNOS基因启动子的转录活性;(2)在LPS刺激下,p38 MAPK 4种亚型均能够诱导iNOS启动子的转录活性,其中,p38β所诱导的转录活性最高,p38α的诱导作用亦很明显。结论(1)LPS能够在HEK 293细胞中诱导iNOS基因转录活性;(2)在HEK 293细胞中,p38 MAPK参与了静息时及LPS刺激下对iN-OS基因的转录调控。     

外源性p21WAF1/CIP1基因转染对人胶质瘤细胞生长和细胞周期的影响

目的：探讨外源性p21^WAF1／CIP1基因对胶质瘤细胞生长和细胞周期的影响．方法：采用脂质体介导法将p21^WAF1／CIP1基因质粒转导人人胶质瘤细胞系SHG44细胞，然后用电镜、流式细胞仪等技术对细胞形态、生长及细胞周期进行观察。结果：酶切电泳和斑点杂交显示p21^WAF1／CIP1基因成功的转染至SHG44细胞。转染p21^WAF1／CIP1基因的细胞光镜及电镜下可见凋亡产生；生长速度明显慢于亲代细胞，其倍增时间约是对照细胞的二倍；细胞周期发生明显改变，G1期明显增多，S期明显减少．结论：外源性p21^WAF1／CIP1基因对SHG44细胞的生长和细胞周期有明显的抑制作用，并可诱导细胞凋亡产生。     

Adenocarcinoma of the Ampulla of Vater: T-Stage,Chromosome 17p Allelic Loss,and Extended Pancreaticoduodenectomy are Relevant Prognostic Factors

Objective To evaluate the prognostic significance of different clinico-pathological and molecular factors, and to compare survival after standard and extended pancreaticoduodenectomy (PD) in ampulla of Vater adenocarcinoma (AVAC). Summary Background Data There are discordant data on factors affecting prognosis, and hence therapeutic choices, in AVAC. Patients and Methods Clinical-pathological factors were evaluated in 59 patients, subjected to PD for AVAC; in 42 subjects information on chromosome 17p and 18q allelic losses (LOH) and microsatellite instability (MSI) was also available. The association between survival and type of PD was investigated in the 25 patients operated between 1990 and 2001 (16 standard and nine extended). Results The overall 5- and 10-year tumor-related survival rates were 46% and 33%, respectively. Sixteen patients had T-stages 1–2, 14 T-stage 3, and 29 T-stage 4 cancers. Chromosome 17p and 18q LOH were detected in 23 (55%) and 15 cases (36%), respectively, and in 12 cases (29%) coexisted. Five cases were MSI-positive (12%). At univariate analysis, poor survival was associated with cancer ulceration (P = 0.051), poor differentiation (P = 0.008), T-stage 4 (P < 0.001), nodal metastases (P = 0.004), chromosome 17p (P < 0.001) and 18q LOH (P = 0.002), and absence of MSI (P = 0.009). At multivariate analysis, only T-stage (P = 0.002) and 17p LOH (P = 0.001) were independent predictors of survival. All patients with MSI-positive cancers were long-survivors (>12 yrs), whereas only 30% of MSI-negative cancer patients survived at 5 years. Extended pancreaticoduodenectomy was associated with a 3-year disease-related survival higher than standard resection (83% vs 31%; P = 0.018). Conclusion MSI and chromosome 17p status allow to better define prognosis within ampullary cancers at the same stage. Surgery alone resulted curative in MSI-positive cancer patients, whereas it was inadequate in patients showing allelic losses, who might benefit from adjuvant therapy. In this observational study, extended PD was associated with increased survival compared to standard procedures. Presented at the 2006 Annual Meeting of the American Hepato-Pancreato-Biliary Association, Miami Beach, Florida, March 9–12, 2006     

Expression of p27 and p53: comparative analysis of uterine carcinosarcoma and endometrial carcinoma

The aim of the study was to assess both p27 and p53 expression in the stromal and epithelial component of carcinosarcoma and to assess if their expression in the latter is different than in endometrial carcinoma. Immunohistochemical staining for p27 and p53 was performed on paraffin-embedded tissue blocks of 18 uterine specimens with carcinosarcoma and their expression assessed. Their expression in the epithelial element was also compared to that in 35 paraffin-embedded tissue blocks of endometrial endometrioid carcinoma. Reduced p27 expression was observed in a similarly high proportion of the stromal (77.8%) as well as of the epithelial component (66.7%) of carcinosarcoma. Although statistically not significant, the proportion of reduced p27 expression in endometrial carcinoma (85.7%) was higher than in the epithelial element of carcinosarcoma. The percentage of p53 overexpression in both elements of carcinosarcomas and in endometrial carcinomas was low and also similar (27.8 and 20.0%, respectively). Our results indicate that reduced p27 expression is common and p53 overexpression is infrequent in carcinosarcoma. Their similar rates of expression in the stromal and epithelial elements of the tumor support the contention of a monoclonal origin of carcinosarcoma. Unexpectedly, reduced p27 expression is more common in endometrial carcinoma than in the epithelial element of carcinosarcoma, in spite of the less favorable prognosticators and outcome in the latter. Further studies of p27 expression in carcinosarcoma are indicated to establish its clinical value in this aggressive malignancy.     

小鼠耳蜗核因子-κB p50免疫组化定位

目的 了解核因子-κB(NF-κB)在耳蜗的表达情况.方法 小鼠皮下注射卡那霉素(KA),鼓室注射脂多糖(LPS),以注射等量生理盐水作为对照,用多克隆兔抗体NF-κB p50免疫组化染色,观察NF-κB p50在小鼠耳蜗的表达.结果 NF-κB p50主要定位在耳蜗的螺旋缘、盖膜、血管纹、螺旋韧带、螺旋神经节和神经纤维.整个耳蜗各圈均可观察到免疫反应,而螺旋韧带、盖膜、螺旋突、螺旋神经节和神经纤维均可观察到较强的免疫反应,Corti器的免疫反应比上述结构要弱一些.Corti器的内、外毛细胞,内、外柱细胞,Deiter细胞和Claudious细胞均可观察到NF-κB p50的免疫反应,这些免疫反应在内沟细胞、Hensen细胞和Claudious细胞较弱,而在螺旋神经节可观察到较强的免疫反应,内、外毛细胞核未见免疫反应.结论 注射脂多糖和卡那霉素可使NF-κB p50在小鼠耳蜗产生急性反应.