人胰腺癌中bcl—2基因和蛋白的过度表达

目的：分析胰腺癌中bcl-2基因的mRNA转录和蛋白翻译，探讨其在胰腺癌 发生、发展中的作用。方法：以50例胰腺癌组织、15例胰腺癌转移灶组织和10例慢性胰腺炎为研究对象，采用免疫组织化学（SP法）和原位分子杂交法检测bcl-2的表达。结果：Bcl-2蛋白表达与患年龄、性别、TNM分期无关，与肿瘤分化程度、有无转移病灶有关。在胰腺癌组织中的表达低于胰腺炎组织，但明显高于转移灶。免疫组织化学与原位杂交技术检测胰腺癌组织中的bcl-2基因表达结果一致。结论：Bcl-2蛋白可能作用于胰腺细胞癌变阶段，并与胰腺导管上皮癌变有关；bcl-2基因在mRNA和蛋白水平表达是相对稳定的。     

甲状腺癌组织中BAG-1、bcl-2蛋白表达及临床意义

目的 探讨BAG-1、bcl-2在甲状腺癌发生、发展中的作用。方法采用免疫组化法检测48例甲状腺癌、11例甲状腺腺瘤组织中BAG-1、bcl-2蛋白的表达情况，并进行相关性分析。结果BAG-1、bcl-2蛋白在甲状腺癌中的表达率（66．7％、75．0％）均高于甲状腺腺瘤（18．2％、27．3％）：甲状腺癌中两基因的表达呈正相关，BAG-1表达水平在甲状腺癌组织学分型、临床分期及淋巴结转移均有相关性。结论BAG-1特异性表达于甲状腺癌组织中，其可能与bcl-2协同参与甲状腺癌的发生、发展；BAG-1表达状态可作为评估甲状腺癌生物学行为和预后的重要参考指标。     

Smac/DIABLO和bcl-2在胰腺癌中的表达及意义

目的探讨线粒体促凋亡因子Smac/DIABLO和凋亡抑制基因bcl-2的表达情况及其与胰腺癌患者临床病理特征的关系。方法应用荧光实时定量RT-PCR及免疫组织化学技术分别检测正常胰腺组织、胰腺癌组织Smac/DIABLO及bcl-2的表达。结果 Smac/DIABLO mRNA在正常胰腺组织中的表达高于胰腺癌(P<0.05)。在胰腺癌组织中,Smac/DIABLO mRNA在Ⅰ～Ⅱ期患者癌组织标本中的表达明显高于Ⅲ～Ⅳ期(P<0.05)。正常胰腺组织中导管上皮细胞bcl-2染色阳性8例(80.00%)。48例胰腺癌bcl-2蛋白阳性表达35例(72.92%),低于正常胰腺组织阳性表达率(80.00%),但二者无统计学差异(P>0.05)。结论 Smac/DIABLO表达的降低与胰腺癌的发生、发展可能密切相关。     

多原发癌组织中bcl-2表达状况的研究

国内对多原发癌的研究相对较少,尚未见有对多原发癌分子病理学研究的报道[1～3].以往的研究表明,bcl-2基因的过表达在许多单发肿瘤的发生发展中起很重要作用[4～6],本实验应用免疫组织化学方法对多原发癌组织中的bcl-2蛋白表达进行检测.     

BTG-2基因在人胰腺癌中的表达及其与胰腺癌细胞增殖和凋亡的关系

目的探讨BTG-2基因在人胰腺癌中的表达及与胰腺癌细胞增殖和凋亡的关系。方法应用免疫组化、原位杂交等技术,对32例胰腺癌及癌旁组织中BTG-2基因进行检测。结果BTG-2基因在胰腺癌旁组织中相对含量为0.76±0.17,在胰腺癌组织中相对含量为0.58±0.16(P<0.01);PCNA指数 - 的胰腺癌细胞中BTG-2相对含量为0.90±0.12,PCNA指数 以上的胰腺癌细胞中BTG-2相对含量为0.66±0.12(P<0.01);BTG-2基因高表达的标本中可见少量的凋亡细胞,而低表达和中等表达的胰腺癌组织中未见明显凋亡细胞。结论BTG-2基因的低表达与胰腺癌的发生发展相关。     

BTG-2基因在人胰腺癌中的表达及其与胰腺癌细胞增殖和凋亡的关系

目的探讨BTG-2基因在人胰腺癌中的表达及与胰腺癌细胞增殖和凋亡的关系.方法应用免疫组化、原位杂交等技术,对32例胰腺癌及癌旁组织中BTG-2基因进行检测.结果 BTG-2基因在胰腺癌旁组织中相对含量为0.76 ± 0.17,在胰腺癌组织中相对含量为0.58 ± 0.16(P < 0.01);PCNA指数 + ～ + + 的胰腺癌细胞中BTG-2相对含量为0.90 ± 0.12,PCNA指数 + + + 以上的胰腺癌细胞中BTG-2相对含量为0.66 ± 0.12(P < 0.01);BTG-2基因高表达的标本中可见少量的凋亡细胞,而低表达和中等表达的胰腺癌组织中未见明显凋亡细胞.结论 BTG-2基因的低表达与胰腺癌的发生发展相关.     

Survivin在肝细胞肝癌中的表达以及与Ki-67和bcl-2的关系

为研究Survivin在肝细胞癌(HCC)发病机制中所起的作用,用原位杂交和免疫组织化学的方法检测了HCC、肝硬化、正常肝组织Survivin mRNA表达及蛋白表达。同时,选择Ki-67和bcl-2,进行蛋白水平的检测,研究Survivin与两者的关系,从而在多基因变化与相互关系、增殖与凋亡的角度来探讨HCC的发生、发展机制。     

氟对软骨细胞凋亡因子bcl-2和Bax蛋白表达的影响

目的 观察氟对体外培养软骨细胞中凋亡相关因子bcl-2和Bax蛋白表达的影响,探讨氟在致软骨细胞凋亡中的作用机制.方法 采用软骨细胞体外培养方法,原代培养乳鼠关节软骨细胞,传第3代后按染氟剂量不同分为0(对照)、5、20、40mg/L组,培养10 d后,透射电镜下观察软骨细胞的超微结构改变,采用Western印迹法检测软骨细胞bcl-2和Bax蛋白表达.结果 透射电镜下,对照组和5 mg/L组软骨细胞呈球形,粗面内质网发达,线粒体膜性结构完整;20、40 mg/L组软骨细胞内可见脂滴增多,胞质内出现大量空泡类物质,细胞内膜结构不清,部分细胞出现核固缩.20、40 mg/L组软骨细胞bcl-2蛋白表达(0.626±0.042、0.531±0.039)较对照组(0.876±0.035)明显降低(P均＜0.01);而Bax蛋白表达(0.966±0.047、1.289±0.156)较对照组(0.642±0.050)明显增加(P均＜0.01).5 mg/L组软骨细胞bcl-2、Bax蛋白表达(0.885±0.065、0.657±0.045)与对照组比较,差异无统计学意义(P均＞0.05).此外,40 mg/L组与20 mg/L组比较,bcl-2、Bax蛋白表达差异有统计学意义(P均＜0.01).结论 染氟20、40 mg/L可对软骨细胞超微结构造成损伤,其通过减少抑凋亡因子bcl-2的表达和增加促凋亡因子Bax的表达,从而产生促进软骨细胞凋亡的作用.

Abstract：

Objective To study the effect of fluoride on the expression of bcl-2 and Bax in chondrocyte in vitro, and investigate the mechanism of action of chondrocyte apoptosis induced by fluoride. Methods Articular chondrocytes of neonate rat were cultured in vitro and treated with 0(control),5,20,40 mg/L of fluoride,respectively, for 10 days. Then observed the u]trastructure of chondrocytes under eletronicmicroscope, and tested the expression of bcl-2 and Bax in chondrocyte in different groups by Western blotting. Results Abundant rough endoplasmic reticulums (RERs) and complete structure of mitochondria membranes were presented in globular chondrocytes in the control group and 5 mg/L group; but more lipid droplets and vacuoles were seen in the cytoplasm, and the structure of intracellular membranes became incomplete, and some shrieked chromatin and pyknosis were seen in the chondrocytes of the 20,40 mg/L groups. The expression of bcl-2 markedly decreased in 20 mg/L group(0.626 ± 0.042) and 40 mg/L group(0.531± 0.039) compared to the control group(0.876 ± 0.035,all P ＜ 0.01 ). And the expression of Bax significantly increased in 20 mg/L group(0.966 ± 0.047) and 40 mg/Lgroup ( 1 .289 ± 0.156) compared to the control group(0.642 ± 0.050, all P ＜ 0.01). But there was no statistical significant difference of the expression of bcl-2 or Bax between 5 mg/L group(0.885 ± 0.065,0.657 ± 0.045) and control group (all P ＞ 0.05 ). However there were statistical differences of expressions of bcl-2 and Bax between 20 and 40 mg/L groups(all P ＜ 0.01 ). Conclusions Twenty and 40 mg/L fluoride can cause damage to the ultrastructure of chondrocyte, and fluoride possibly promotes chondrocyte apoptosis by reducing the expression of antiapoptotic factor bcl-2 and increasing the expression of Bax.     

胰腺癌相关基因研究进展

近年来，大量研究表明，肿瘤是多种遗传基因改变累积的结果，其中癌基因和抑癌基因的平衡失调是导致癌细胞无限增殖的重要原因。胰腺的恶性肿瘤分为腺癌、内分泌肿瘤及非上皮恶性肿瘤三类，其中90％以上为腺癌，预后极差。在分子水平上研究胰腺癌的发生和发展，成为胰腺实验外科研究的热点。现将胰腺癌基因研究的进展情况介绍如下。     

胰腺癌环氧合酶-2表达的意义

目的研究胰腺癌中COX-2表达的意义．方法应用免疫组化SP法检测82例胰腺癌、22例慢性胰腺炎、9例胰腺良性肿瘤、15例正常胰腺组织和2株人胰腺癌细胞中COX-2的表达,然后比较胰腺癌组织COX-2表达与胰腺癌患者临床病理特征的关系。结果2株人胰腺癌细胞COX-2表达均阳性。70．7％（58／82）胰腺癌组织可见COX-2表达,而正常胰腺组织只有6．7％（1／15）呈微弱表达。COX-2在4种组织中的表达程度有显著性差异（P〈0．001）。胰腺癌组织COX-2表达显著高于其他3种组织（P〈0．05）。胰腺癌组织COX-2表达水平的高低与胰腺癌的临床病理特征无关（P〉0．05）。结论COX-2蛋白的检测对胰腺癌的诊断及其与胰腺良性肿瘤、慢性胰腺炎的鉴别诊断有帮助。胰腺癌组织COX-2表达不能作为预测患者预后的指标。     

Variants of bcl-2 specific siRNA for silencing antiapoptotic bcl-2 in pancreatic cancer

BACKGROUND AND AIMS: Pancreatic cancer remains a devastating diagnosis with only limited therapeutic options. Specific inhibition of expression of target genes has become possible using small interfering (si) RNAs. We therefore investigated how far siRNA specific for bcl-2 may serve as a therapeutic option for pancreatic cancer in vitro and in vivo. METHODS: siRNAs targeting two different regions in the bcl-2 gene were transfected to YAP C and DAN G pancreatic carcinoma cells and human foreskin fibroblasts. Permutations were generated by changing 3' and 5' overhangs and varying the length of the paired RNA duplex. Transfection efficacy was determined using FITC labelled siRNAs and fluorescence microscopy. Cell survival and apoptosis were quantified at 24-120 hours. Pancreatic cancer xenografts in male nude mice were treated intraperitoneally with siRNAs daily for 24 days. siRNA pharmacokinetics in vivo were assessed using radioactively labelled siRNAs. Total protein and RNA were extracted for western Blot analysis and quantitative polymerase chain reaction. RESULTS AND CONCLUSIONS: Bcl-2 specific siRNAs specifically inhibited expression of the target gene in vitro and in vivo. Antiproliferative and proapoptotic effects were observed in tumour cells but not in fibroblasts or non-malignant tissues. siRNA permutations and diverse overhangs influenced gene silencing efficacy. siRNA was quickly distributed to all organs and excreted via the kidney and liver. Bcl-2 specific siRNA is a promising adjunctive treatment for pancreatic carcinoma.     

Clinicopathological significance of Bcl-2 and Bax protein expression in human pancreatic cancer

AIM: To assess the clinicopathological significance of the expression of the apoptosis-inhibitory Bcl-2 protein (pBcl-2) and the apoptosis-promoting Bax protein (pBax) in human invasive ductal carcinomas (IDCs) of the pancreas. METHODS: Fifty-nine surgical specimens of IDCs of the pancreas were stained immunohistochemically to detect pBcl-2 and pBax expressions whose correlation to tumor classification, staging, and prognosis was analyzed by univariate and multivariate analyses. RESULTS: The expression of pBcl-2 and pBax was detected in 21 of 59 (35.6%) and in 29 of 59 (49.2%) patients with IDCs of the pancreas, respectively. Neither pBcl-2 nor pBax alone was correlated to TIMM staging and differentiation degree of IDCs of the pancreas according to univariate analysis. By Mantel-Cox test, the median survival time after surgery for pBcl-2(+) and pBcl-2(-) groups were 14.3 and 7.3 mo, respectively (x~2=9.357, P=0.002) and that for pBax(+) and pBax(-) groups were 12.9 and 10.2 mo, respectively (x~2=0.285, P＞0.05). Contingency coefficient between pBcl-2 and pBax expression was 0.298, indicating that there was correlation between them (x~2=5.74, P＜0.05). The median survival time after surgery for pBcl-2(+)pBax(+) and pBcl-2(+)pBax(-) groups were 14.3 and 14.1 mo, respectively, and that for pBcl-2 (-)pBax(+) and pBcl-2(-)pBax(-) groups were 5.9 and 9.9 mo, respectively. There was a significant difference between pBd-2(+)pBax(+) and pBd-2(-)pBax(+) (x~2=5.06, P＜0.05), such was the case for pBcl-2(+)pBax(+) and pBd-2(-)pBax(-) (x~ 2=7.18, P＜0.01). Cox proportional hazards model for multivariate analysis was applied, indicating that pBcl-2, TNM staging, age and pBax were high risk factors of post-surgical survival time. CONCLUSION: Both pBcl-2 and pBax have high expression in IDCs of the pancreas, indicating that co-expression of pBcl-2 and pBax is a good indicator of favorable prognosis in IDCs of the pancreas.     

胰腺癌细胞基因组范围内的基因缺失

目的:研究胰腺癌细胞基因组范围内的纯合性缺失和杂合性缺失(loss of heterozygosity, LOH).方法:应用高密度单核苷酸多态性芯片和专用分析软件,检测17种胰腺癌细胞株基因组范围内的纯合性缺失和LOH,并筛选可能与胰腺癌发生、发展有关的基因区域,用PCR验证纯合性缺失.结果:经过PCR验证,26个区域确实为纯合性缺失,芯片的准确度为83.9%(26/31).这些缺失区域中,平均每个区域只涉及1.29个基因.每一种细胞都有不同程度的LOH;不同染色体臂出现LOH频率不同,出现频率最高的为染色体臂9p和18q,均为94.1%.结论:胰腺癌全基因组范围内出现多处LOH和纯合性缺失,这些区域可能含有新抑癌基因.     

Clinicopathological significance of p53 and mdm2 protein expression in human pancreatic cancer

AIM: To study the clinicopathological significance of p53 and mdm2 protein expression in human pancreatic cancer. METHODS: To investigate the expression of p53 and mdm2 in pancreatic cancer by immunohistochemistry, and the relationships between the p53 and mdm2 protein expression and clinicopathological parameters in pancreatic cancer. RESULTS: The positive expression of p53 protein was found in 40 of 59 patients (67.8%) and that of mdm2 protein in 17 of 59 patients (28.8%). No obvious relationships were found between p53 as well as mdm2 expression and sex, tumor site,TNM staging and histological differentiation. p53 expression was increased in patients younger than 65 years old, while mdm2 had no relationship with age. The survival time of the patients with the positive expression of p53 and mdm2 proteins was obviously shorter than the other groups. CONCLUSION: Both p53 and mdm2 presented relatively high expression in human pancreatic cancer.The overexpression of p53 and mdm2 might reflect the malignant proliferation of pancreatic cancer and their co-expression might be helpful to evaluate the prognosis of the patients with pancreatic cancer.     

B-CLL and bcl-2 gene

Most of human follicular lymphomas (approximately 90% in U.S.A. or approximately 30% in Japan) possess the t(14; 18) chromosome translocation that directly involves the IgH locus on chromosome 14 and the bcl-2 gene on chromosome 18. The t(14; 18) chromosome translocation occurs nearly exclusively at two hot spots, a major breakpoint clustering region (mbr) within the 2nd exon noncoding region and the minor breakpoint clustering region (mcr) within the 3' flanking region of the bcl-2 gene. Here we show that the rearrangement of the bcl-2 gene occurs in a significant fraction (approximately 10%) of B-CLL. All of the rearranged bcl-2 genes were juxtaposed with Ig lambda or Ig kappa genes, implying that the bcl-2 gene is preferentially linked to the IgL genes in CLL.     

胰腺癌胆囊收缩素受体蛋白表达的研究

目的探讨胆囊收缩素受体在正常胰腺组织、胰腺癌组织、胰腺癌细胞株SW1990中的表达,以明确胆囊收缩素受体与胰腺癌的关系.方法应用免疫组织化学法和免疫细胞化学法分别检测正常胰腺组织、胰腺癌组织、胰腺癌细胞株SW1990中胆囊收缩素受体A、B亚型的表达.结果在正常胰腺组织中缺乏胆囊收缩素受体A、B亚型的表达;在胰腺癌组织、人胰腺癌细胞株SW1990中也不表达胆囊收缩素受体A亚型,但可见胆囊收缩素受体B亚型的表达;胆囊收缩素受体在癌细胞株SW1990中的表达主要位于细胞膜上,细胞质内也可见.结论胆囊收缩素受体B在胰腺癌组织、腺癌细胞株SW1990中表达;胆囊收缩素受体B亚型主要位于细胞膜上;胆囊收缩素受体B亚型在胰腺癌细胞生长中可能发挥重要作用.     

ERBB2在胃癌组织中过表达的研究

目的研究癌基因ERBB2在胃癌组织中过表达情况,并与临床资料进行对比观察。方法65例不同病理类型的胃癌组织及25例正常对照组织进行ERBB2过表达的免疫组织化学染色,其表达结果与临床资料进行对比分析。结果65例胃癌中ERBB2过表达者8例,占12.3%（8/65）;而癌旁正常胃组织未见ERBB2过表达（P〈0.01）。其中肠型胃癌7例,占该型胃癌的16.3%（7/43）;弥散型胃癌1例,占该型胃癌的4.5%（1/22）;两者相差显著（16.3%vs4.5%,P〈0.01）。ERBB2过表达与胃癌TNM分期相关联,TNM分期越差者,其阳性率越高。结论ERBB2在胃癌中存在过表达,且在不同病理类型的胃癌中所起作用可能不同。