HER-2、PCNA、Bcl-2、Bax在乳腺癌组织中的表达与预后的关系

背景与目的:临床上在早期将具有高危因素及潜在转移危险的乳腺癌患者筛选出来并进行针对性的治疗是提高乳腺癌生存率、减少复发和转移的关键。本研究探讨人表皮生长因子受体2(human epidermal receptor 2,HER-2)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、Bcl-2、Bax的表达与乳腺癌患者预后的关系。方法:应用免疫组织化学EnVision二步法检测50例乳腺癌根治术后标本中HER-2、PCNA、Bcl-2、Bax蛋白的表达,结合5年随访资料分析上述指标与患者预后的关系。结果:乳腺癌组织中HER-2、PCNA、Bcl-2、Bax的阳性率分别为44.0%(22/50)、82.0%(41/50)、48.0%(24/50)、54.0%(27/50)。患者的中位生存期为62个月,5年总生存率为82.0%(41/50),无瘤生存率为38.0%(19/50)。单因素生存分析显示HER-2和Bax是乳腺癌复发的影响因素(P<0.001),多因素Cox模型分析显示HER-2和Bax是影响乳腺癌复发的独立因素(P<0.05和P<0.01),Bcl-2和Bax是影响生存期的独立因素(P<0.05)。结论:影响乳腺癌患者预后的独立因素有HER-2、Bcl-2、Bax,其中HER-2、Bax表达是预后不良的指标,Bcl-2表达是预后良好的指标。

浸润性膀胱癌Bcl-2、Her-2表达与保存膀胱术后放化疗疗效的相关性研究

根治性膀胱全切术是浸润性膀胱癌的标准治疗方式，而近年研究认为，一部分浸润性膀胱癌患者采用保存膀胱手术的综合治疗，在保留膀胱功能的同时可达到与膀胱全切术相似的生存率，长期生存患者的膀胱功能良好，为了筛选出可从保存膀胱术后放化疗中获益的患者，实现浸润性膀胱癌的个体化治疗，

Bcl-2与HER-2反义寡核苷酸联合转染对人舌鳞癌Tca8113细胞的干预作用

目的探讨Bcl-2与HER-2基因反义寡核苷酸(ASODN)联合转染对人舌鳞癌Tca8113细胞增殖的干预作用及其分子机制。方法实验分成正常对照组、Bcl-2正义干预组、HER-2正义干预组、Bcl-2反义干预组、HER-2 反义干预组、Bcl-2与HER-2基因反义寡核苷酸联合干预组,共6组。脂质体转染后不同时间分别进行电镜观察、 RT-PCR检测,以观察不同条件处理后各组细胞的凋亡、Bcl-2与HER-2基因表达有无差别。结果 Bcl-2与HER-2 基因反义寡核苷酸联合干预组,转染后出现凋亡小体和各期凋亡细胞;Bcl-2基因表达下降36％,HER-2基因表达下降51％。联合转染作用优于单反义基因转染作用。结论 Bcl-2与HER-2基因反义寡核苷酸联合转染能够促进肿瘤细胞凋亡并下调Bcl-2与HER-2基因的表达。

Correlation between expression of gastrin, somatostatin and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma

AIM: To explore the correlation between expression of somatostatin (SS), gastrin (GAS) and cell apoptosis regulation gene bcl-2/bax in large intestine carcinoma.METHODS: Sixty-two large intestine cancer tissue samples were randomly and retrospectively selected from patients with large intestine carcinoma. Immunohistochemical staining for bcl-2, bax, GAS, SS was performed according to the standard streptavidin-biotin-peroxidase (S-P) method.According to the semi-quantitative integral evaluation, SS and GAS were divided into three groups as follows. Scores1-3 were defined as the low expression group, 4-8 as the intermediate expression group, 9-16 as the high expression group. Bax and bcl-2 protein expressions in different GAS and SS expression groups of large intestine carcinoma were assessed.RESULTS: The positive expression rate of bax had a prominent difference between SS and GAS high, intermediate and low expression groups (P＜0.05, x2ss = 9.246; P＜0.05,x2GAS = 6.981). The positive expression rate of bax in SS high (80.0%, 8/10) and intermediate (76.5%, 13/17)expression groups was higher than that in low expression group (40.0%, 14/35) (P＜0.05, x2high vs low = 5.242; P＜0.05,x2middle vs low = 6.097). The positive expression rate of bax in GAS high expression group (27.3%, 3/8) was lower than that in low expression group (69.4%, 25/36) (P＜0.05,x2 = 4.594). However, bax expression in GAS intermediate expression group (46.7%, 7/15) was lower than that in low expression group, but not statistically significant. The positive expression rate of bcl-2 had a prominent difference between SS and GAS high, intermediate and low expression groups (P＜0.05, x2ss = 7.178; P＜0.05, x2GAS = 13.831). The positive expression rate of bcl-2 in GAS high (90.9%, 10/11)and intermediate (86.7%, 13/15) expression groups was higher than that in low expression group (44.4%, 16/36)(P＜0.05,x2high vs low = 5.600; P＜0.05, x2 middle vs low = 7.695).However, the positive expression rate of bcl-2 in SS high (40.0%, 4/10) and intermediate (47.1%, 8/9) expression groups was lower than that in low expression group (77.1%, 27/35)(P＜0.05, x2 high vs low = 4.710; P＜0.05, x2 middle vs low = 4.706).There was a significant positive correlation between the integral ratio of GAS to SS and the integral of bcl-2 (P＜0.01,r=0.340). However, there was a negative correlation between the integral ratio of GAS to the SS and bax the integral of (P＜0.05, r = -0.299).CONCLUSION: The regulation and control of gastrin,somatostatin in cell apoptosis of large intestine carcinoma may be directly related to the abnormal expression of bcl-2, bax.

Altered Oncogene Activity Contributes to Compensation for Antisense Suppression of Bcl-2 and Tumor Resistance

Antisense oligonucleotides (oligos) have targeted growth regulatory proteins in prostate cancer models. To identify compensatory alterations in the expression of non-targeted genes we evaluate mono- and bispecific oligos targeting and equally suppressing the expression of the apoptosis inhibitory protein bcl-2. Bcl-2 is chosen because oligos directed towards it have entered clinical trials to restore apoptosis in cancer patients. Treated LNCaP cells compensate for the diminished bcl-2 by suppressing caspase-3 (an apoptosis promoter) while enhancing expression of AKT-1 (another apoptosis inhibitor), androgen receptor (AR) and its (p300 and IL-6) coactivators. Additional proteins are enhanced including PD-1, its ligand PD-L1 (immune checkpoint blockade markers) and fas-ligand, which activate apoptosis through the signal transduction, along with suppressor protein p53, polymerase transcription mediator MED-12 and signal transducer STAT-3. These alterations in expression may contribute to a greatly enhanced expression of the proliferation marker KI-67. This suggests that therapeutic approaches to restore apoptosis through suppression of bcl-2 lead to an altered expression in non-targeted genes involving apoptosis, androgen sensitivity, transcriptional activity and immune responsiveness, leads to an increase in proliferation (and a more androgen driven aggressive phenotype). In this study we evaluate the expression of two oncogenes (v-myc and K-ras) and find a large and significant enhancement of v-myc activity, which is produced by oligos targeting bcl-2 at the 5’ position. For K-ras, although significant suppression is produced by the bispecific targeting bcl-2 at the 3’ position, the percent change is relatively small compared with other compensatory alterations we have measured, and much less than in v-myc. Therefore, for the two oncogenes being evaluated, only increased v-myc activity is probably large enough to contribute to increased tumor aggressiveness in compensation for bcl-2 suppression.

The expression of HER-2/neu gene in colon cancer tissues and its clinical significance

Objective：The article aims to detect the expression of HER-2/neu gene in colon cancer tissues and adjacent tissues, to analyze the relationship between dif erent pathologic types and clinical features, also to invest the distribution of patients with positive expression of HER-2 gene. Methods：The expression of HER-2 gene in the 223 samples with colon can-cer was detected by immunochemical approach. The expression of HER-2 gene in colon cancer tissues and adjacent tissues and dif erent pathologic types was analyzed byχ2 test. The correlation between the expression of HER-2 gene and clinical features was analyzed by Spearman. Results：The number of positive expression of HER-2 gene in colon cancer tissues and adjacent tissues were 74 and 0 respectively, the dif erence has statistical significance. The number of papil ary or tubular adenocarcinoma was 182, among them, 60 cases were positive expression. The number of mucinous adenocarcinoma was 41, among them, 14 cases were positive expression. The expression of HER-2/neu gene has no correlation with sex, age, the maximum diameter, general classification, degree of dif erentiation and depth of invasion, which has no statistical significance. However, the expression of HER-2/neu gene has correlation with metastasis of lymph node and Dukes stage, which has statistical significance. The expression of HER-2/neu gene was positive correlation with metastasis of lymph node and Dukes stage. The correlated coef icient index was 0.320 and 0.320 respectively. In the 74 patients with positive expression of HER-2 gene, 59.4%of them were 60-74 years old. And there was 97.3%of the patients without family history of adenocarcinoma. Conclusion：The expression of HER-2/neu gene in colon cancer tissues was higher than in adjacent tissues. The expression of HER-2/neu gene has no correlation with sex, age, the maximum diameter, general classification, degree of dif erentiation and depth of invasion, but has correlation with metastasis of lymph node and Dukes stage. The expression of HER-2/neu gene was positive correlation with metastasis of lymph node and Dukes stage. The expression of HER-2/neu gene with age of 60-74 years old and without family history of adenocarcinoma was higher than other groups.

Effects of Bcl-2 Gene Interference on the Apoptosis,Proliferation and Progesterone Secretion of Goose Follicular Granulosa Cells

Based on published sequences for chicken Bcl-2,three siRNAs（small interfering RNA）were designed,and expression vectors were constructed and transfected into goose granulosa cells cultured in vitro.Bcl-2 protein,apoptosis and proliferation of granulosa cells,48 h after the transf ection,were analyzed by flow cytometry,and progesterone（P）secreted into the culture medium was measured by radioimmunoassay.In addition,apoptosis and Bcl-2 protein level were assessed in untreated granulosa cells from the four largest preovulatory follicles（F1F4）,the smallest preovulatory follicles（SPF）,small yellow follicles（SYF）and atretic follicles.The highest level of Bcl-2 protein was observed in granulosa cells from SPF,and levels in cells from healthy follicles were significantly higher than those of atretic follicles（P【0.05）.Bcl-2 protein levels in cells subjected to RNAi were significantly lower than those of controls（P【0.05）,while apoptosis indices（AI）,proliferation indices（PI）and P secretion in the RNAi treatments were higher than those of controls（P【0.05）.

Bcl-2 GENE REARRANGEMENT DETERMINED BY PCR AS AMEAN TO DETECT MINIMAL RESIDUAL DISEASE INMALIGNANT LYMPHOMAS

Objective: To develop a sensitive method to detect minimal residual disease and to elucidate the significance of bcl-2 gene rearrangement in diagnosis and treatment of malignant lymphoma. Methods: Using polymerase chain reaction (PCR) to detect bcl-2 gene rearrangement and using serial dilution method to define the sensitivity of PCR. Results: In 9 different malignant lymphoma cell lines, Su-DHL-4 and Su-DHL-6 were shown bcl-2(MBR)/JH rearrangement, the sensitivity of PCR was 1:105. In 16 patients with follicular lymphoma, the peripheral blood and bone marrow were PCR positive in 4 cases both at initial diagnosis and after complete remission. Conclusion: Detection of bcl-2 gene rearrangement by PCR provides a sensitive and specific assay of minimal residual disease. It is helpful to improve staging of disease, prognosis and evaluation of the treatment results.

Detection of apoptotic cells and immunohistochemical study of bcl-2 and p53 gene protein in primary gastric mucosa-associated lymphoid tissue (MALT) lymphoma

To identify the apoptotic cells in gastric MALT lymphoma and its relationship between bcl-2 and p53 gene expression. Methods: TdT-mediated dUTP biotin Nick End labeling (TUNEL) and immuno-histochemistry ABC method were used to display apoptotic cells and the gene protein expression of bcl-2 and p53 independently. Results: Apoptotic indices (AI) in high-grade MALT lymphomas were significantly higher than in mixed-grade group and low-grade group (P<0.05). Bcl-2 was expressed in 83% of low-grade tumors, 61.6% of the median-grade tumors and 43.7% of high-grade tumors. An inverse correlation was observed between the expression of bcl-2 and apoptotic indices. Only 27 cases were p53 positive. The frequency of p53 positivity was significantly increased as the histologic grade advanced (P<0.05). There was also an inverse correlation between the expression of bcl-2 and p53. Conclusion: Apoptosis may be important in tumors development and transmission. p53 and bcl-2 were important regulatory genes of apoptosis and may be associated with transformation from low- grade to high-grade lymphomas.

Study on the Regulation of Bcl-2 Gene on Rat Spermatogenic Cells Apoptosis in Transcription Level

Objective To detect the change of Bcl 2 gene expression in the apopototic process of spermatogenic cells in rat with vasoligation and vasostomy, and to find out the relationship between the transcription of Bcl 2 and the apoptosis of spermatognic cells Materials & Methods Sixty adult male Sprague Dawley rats in 3 groups were operated with vasoligation and vasostomy. Then hybridization in situ with hypersensitive Bcl 2 RNA probe was used to detect the change of Bcl 2 mRNA. Results The transcription of Bcl 2 gene in spermatogenic cells was obviously inhibited in the vasoligation group compared with that in the control group (P<0.05), and the transcription in the vasostomy group showed no difference from that of the control group. Conclusion Bcl 2 gene has an anti apoptotic effect in rats with vasostomy, and there was a transcriptional regulation of Bcl 2 gene in rat spermatogenic cell during the period of pre vasoligation to post vasoligation and to post vasosotomy.

Silencing of Bcl-2 gene expression by siRNA transfection in- hibits the protective effect of fluvastatin against cell apoptosis in human aortic endothelial cells

Objective To study the protective effect of fluvastatin,one of the HMG-CoA reductase inhibitors (statins),against oxygen radical-induced oxidative damages in human aortic endothelial cell,and the role of Bcl-2 in this protection.Methods Human aortic endothelial cells with or without Bcl-2 siRNA transfection were subjected to 1-100 nM of fluvastatin and 100 la hydrogen peroxide for 24 hours.Bcl-2 mRNA and protein expression were measured by Taqman quantitative PCR and Western blotting.Cell apoptosis was measured by normal and fluorescent microscopy and Cell Death Detection ELISA.Results In the Bcl-2-expressed cells,fluvastatin significantly reversed hydrogen peroxide-induced microscopic apoptosis and apoptotic DNA fragmentation,which were accompanied by a markedly upregulation of Bcl-2 expression by fluvastatin.However,the endothelial protection by fluvastatin was completely lost in Bcl-2 siRNA transfected cells.Conclusion Fluvastatin protects human endothelial cells against oxygen radical-induced cell apoptosis in vitro,and this protection seemed to be mediated in a Bcl-2 dependent pathway.(J Geriatr Cardil 12008;5:33-38)

DETECTION OF BCL-2 GENE MAJOR BREAKPOINT REGION REARRANGEMENT IN HUMAN B-CELL LYMPHOMAS

Objective To investigate the frequency of t(14; 18) in different subtypes of B-cell lymphomas and the ability or the polymerase chain reaction(PCR) to detect this rearrangement in frozen samples. Methods 1o7 cases of B-cell lymphomas were studied uslng DNA extracted from rresh-frozen tissues. The DNA samples were amplified by PCR for bcl-2 MBR/JH. The products of bcl-2/JH rearrangement were hybridized with an internal olignucleotide probe or bcl-2 MBR. Results The rearranged bcl-2MBR/JH gene was detected in 13 of the 25(52. o% ) follicular center lymphomas, according to REAL classification: 8 of 11 (72. 7%) grade 1, 2 of 5(40. 0%) grade I, and 3 of 90 (33. 3%) grade, 17 of 82(2o. 8%) cases or difruse large B-cell lymphomas were found to have detectable bel-2 MBR/J. rearrangement- Conclusion The rrequency or bcl-2 MBR/JH rearrangement in diffuse large B-cell lymphomas is significantly lower than those in follicular center lympkomas(X2= 9. 28, P <o. oo5), suggesting that bcl2/JH rearrangements occur mainly in follicular center lymphomas. in addition, the result of reconstruction experiments suggest that amplification or bcl-2 MBR/JH rearrangements by PCR is both sensitive and specific for detection of t (14; 18 ) translocation.

抗HER-2抗体chA21对人乳腺癌SKBR3细胞凋亡的诱导作用及其机制

目的探讨抗HER-2工程抗体chA21在体外对高表达HER-2的人乳腺癌SKBR3细胞凋亡的诱导作用及其分子机制。方法采用透射电镜和原位末端标记技术(TUNEL)观察和检测chA21对SKBR3细胞凋亡的诱导,采用免疫细胞化学技术检测凋亡相关基因bc l-2、bax、Fas及caspase-3表达的改变。结果chA21作用72 h,可见SKBR3细胞凋亡,chA21高浓度组(5.4mg/L)凋亡指数显著高于低浓度组(0.2 mg/L)(P<0.01);chA21处理组SKBR3细胞的bax、Fas及caspase-3表达增加,而bc l-2表达及bc l-2/bax比值降低,上述改变在chA21高、低两个浓度组间有显著差异(P<0.01)。结论chA21在体外可诱导SK-BR3细胞凋亡,其分子机制与调节凋亡相关基因bax、bc l-2、Fas及caspase-3的表达有关。

THE EXPRESSION OF BCL-2 GENE AFTER TRANSIENT FOCAL ISCHAEMIA AND THE EFFECT OF MK-801

in order to clarify the pesible role of the bcl-2 gene imolved in the cell death Program,and the relatiouship of glutamate receptors with bcl-2 gene expressin, this study examied the expression of bcl-2 gene protein, the neuronal status of apoptosis and the effects of MK-801 using immunohistochemistry and in situ terminal.labelling methods after 30 min of.middle cerebral artery(MCA) occlusion and followed by 24 h of reperfusion. The presence of bcl-2 gene protein increased in the ipeilateral hemisphere of ischaemis espeially in the MCA territory MK-801 enhanced the expresion of the bcl-2 gene protein. No DNA fragmentation was detected in this experiment. In conclusion. bcl-2 gene activity increased during transient focal ischaemia, and was potentiated by MK MK801, which may be an endogenous protective mechanism .against ischaemic apoptosis. Apoptosis wasnot detected after tranient focal ischoemia. for 30 min rollowed by 24 h of reperfusiou.

bcl-2小分子干扰RNA对人子宫内膜癌RL-952细胞阿霉素敏感性的影响

目的研究靶向bcl-2小分子干扰RNA(siRNA)干扰序列对人子宫内膜癌RL-952细胞阿霉素(DOX)敏感性的影响及相关作用机制。方法 2014年5—8月,将siRNA干扰序列转染人子宫内膜癌RL-952细胞,设3个实验组:空白对照组、bcl-2 siRNA干扰组(转染bcl-2 siRNA干扰序列)及阴性对照组(转染siRNA空载序列),检测bcl-2 siRNA干扰组的转染效率和3组bcl-2表达水平。将细胞随机分成5组,分别为空白对照组、阴性对照组(转染siRNA空载序列)、干扰组(转染bcl-2 siRNA干扰序列)、DOX组(加入5μg/ml DOX)、联合组(转染siRNA干扰序列并加入5μg/ml DOX),检测细胞活力、细胞凋亡率、半胱氨酸蛋白酶(Caspase)-3、Caspase-9活性及细胞色素C水平。结果 siRNA干扰序列转染人子宫内膜癌RL-952细胞的转染效率为(94.6±12.5)%。3组bcl-2表达水平比较,差异有统计学意义(F=8.75,P<0.05)。bcl-2 siRNA干扰组bcl-2表达水平低于空白对照组和阴性对照组(q=3.99、2.87,P<0.01)。5组细胞活力比较,差异有统计学意义(F=42.52,P<0.05)。干扰组、DOX组、联合组细胞活力低于空白对照组(q=7.66、8.64、6.89,P<0.05);联合组细胞活力低于DOX组(q=6.76,P<0.01)。5组细胞凋亡率比较,差异有统计学意义(F=112.34,P<0.05)。干扰组、DOX组、联合组细胞凋亡率高于空白对照组(q=5.78、4.99、6.21,P<0.05);联合组细胞凋亡率高于DOX组(q=4.89,P<0.01)。5组Caspase-3、Caspase-9活性及细胞色素C水平比较,差异有统计学意义(P<0.05)。干扰组、DOX组、联合组Caspase-3、Caspase-9活性及细胞色素C水平高于空白对照组(P<0.05);DOX组、联合组Caspase-3活性高于阴性对照组,干扰组、DOX组、联合组Caspase-9活性高于阴性对照组(P<0.05);联合组Caspase-3活性高于干扰组,DOX组、联合组Caspase-9活性高于干扰组(P<0.05);联合组Caspase-3、Caspase-9活性及细胞色素C水平高于DOX组(P<0.05)。结论靶向bcl-2 siRNA干扰序列可以增强人子宫内膜癌RL-952细胞的DOX敏感性,bcl-2可作为人子宫内膜癌基因治疗的候选靶点。

Selection of Effective Bcl-2 Antisense Oligodeoxynucleotides with Computer Software and Experimental Assay

Objective： To explore and investigate the selection of effective antisense oligodeoxynuleotides with the help of computer and RNAstructure folding software. Methods： Bcl-2 gene was used as the target gene and five antisense oligodeoxynuleotides were designed to be bound to Bcl-2 mRNA optimal secondary structure regions that were predicted free from intramolecular fold or instability of free energy. The five antisense oligodeoxynucleotides were studied with experimental assay of leukemia cells, including cell grow assay with tropan blue exclusion, expression of Bcl-2 protein detected with immunochemistry and flowcytometry, Bcl-2 mRNA content detected with RT-PCR technique, as well as apoptosis observed and determined with morphonological method, electrophoresis and flowcytometry. Results： The results showed that two of the five antisense oligodeoxynucleotides were effective antisense oligodeoxynucleotides, which were able to inhibit cell growth in leukemia, to decrease the level of Bcl-2 mRNA and protein, to induce apoptosis of leukemia cells significantly. Conclusion： The computational prediction of antisense efficacy is faster than other methods and more efficient, which can potentially speed the development of sequences for both research and clinical applications.

P53和Bcl-2蛋白在基底细胞癌中的表达及临床意义

目的探讨P53、B细胞淋巴瘤/白血病-2基因(bcl-2)蛋白在基底细胞癌(BCC)组织中的表达及临床意义。方法用免疫组化链霉素抗生素蛋白-过氧化物酶连结(SP)法检测P53、Bcl-2蛋白在40例BCC(BCC组)组织中的表达,以20例正常皮肤组织作为对照组。结果 P53、Bcl-2蛋白在BCC组表达阳性率分别为55%(22/40)、85%(34/40),在对照组表达阳性率分别为15%(3/20)、10%(2/20)。P53、Bcl-2蛋白在BCC组中的表达明显强于对照组,差异有统计学意义(χ2=6.35、7.45,P<0.01)。结论 P53、Bcl-2蛋白的异常表达与BCC相关,说明其可能在BCC的发生及发展过程中起重要作用。

凋亡蛋白对Bcl-2家族几个基因表达影响

凋亡蛋白(Apoptin)只诱导肿瘤细胞或转化的细胞凋亡,但其作用机制还不十分清楚.从鸡贫血病毒总DNA中克隆出凋亡蛋白基因,构建pcDNA3.1/CT-Apoptin-GFP-TOPO融合表达的载体.用该载体转染HHCC单层传代细胞24 h后,观察到绿色荧光蛋白表达,同时观察到细胞形态呈凋亡特征变化.提取细胞总RNA,按SuperArray芯片操作方法检测到实验组Bcl-2家族中的BimL,Bcl-2,Bax和Bak基因上调表达.Bcl-2家族在细胞凋亡的过程中作用重要.凋亡蛋白诱导HHCC细胞凋亡过程中,促凋亡的BimL,Bax 和Bak基因上调,抗凋亡的Bcl-2基因也上调.可能BimL与Bcl-2竞争性结合导致Bax或Bak释放,最终诱导细胞凋亡.揭示Bcl-2家族的基因参与凋亡蛋白诱导的HHCC细胞凋亡过程.

Relationship between expression and distribution of cyclooxygenase-2 and bcl-2 in human gastric adenocarcinoma

AIM: To explore expression and distribution features of COX-2 and bcl-2 in human gastric adenocarcinoma tissues and to study its biological significance.METHODS: Totally 36 human gastric carcinoma samples were enrolled in this study (cardiac adenocarcinoma 16 cases, distal gastric adenocarcinoma 20 cases). The expressions of COX-2 and bcl-2 in cancerous tissues and corresponding para-cancerous tissues were investigated by immunohistochemistry using COX-2 polyclonal antibody and bcl-2 monoclonal antibody. The normal gastric mucosa tissues were used as control.RESULTS: The expressions of COX-2 and bcl-2 in gastric carcinoma were significantly higher than that in the paracancerous tissues (77.8% vs 47.2%, P＜0.01, 80.56% vs 58.33%, P＜0.05). The expression of COX-2 in cardiac adenocarcinoma was remarkably higher than that in the distal gastric carcinoma (93.8% vs 65.0%, P＜0.01). The expression of COX-2 was mainly localized in the cytoplasm of tumor cells and partly in the nucleus. There is a transition of the COX-2 cytoplasmic positivity to nucleic in tumor cells with the increase of gastric carcinoma pathological grade. Interstitial macrophages, fibroblasts and vascular endothelial cells also expressed COX-2. The tissues with higher expression of COX-2 also expressed high level of bcl-2 protein.CONCLUSION: Abnormal expression pattern of COX-2within the tissues of human gastric cancer is correlated with tumor location and lymph node metastasis. COX-2may regulate expression of apoptosis suppressor gene (bcl-2) through interaction of tumor cells and stromal cells and play an important role in the generation and development of tumors, which will be of great help in developing new methods for antitumor therapy.

Intravenous acid fibroblast growth factor protects intestinal mucosal cells against ischemia-reperfusion injury via regulating Bcl-2/Bax expression

AIM: To detect the effect of acid fibroblast growth factor (aFGF) on apoptosis and gene expression of bax and bcl-2 gene in rat intestine after ischemia/reperfusion (I/R) injury, and to explore the protective mechanisms of aFGF.METHODS: One hundred and eight Wistar rats were randomly divided into sham-operated control group (C)(n = 6), intestinal ischemia group (I) (n = 6), aFGF treatment group (A) (n = 48) and intestinal ischemia reperfusion group (R) (n = 48). In group I, the animals were killed after 45 min of superior mesenteric artery (SMA) occlusion, while in groups R and A, the rats sustained 45 min of SMA occlusion and were then treated with normal saline and aFGF, respectively, sustained 15 min, 30 min, 1, 2, 6, 12, 24, or 48 h of reperfusion, respectively. In group C, SMA was separated, but without occlusion. Apoptosis in intestinal villus was determined with terminal deoxynucleotidyl transferase mediated dUTP-biotin nickend labeling technique (TUNEL). Intestinal tissue samples were taken not only for detection of bax and bcl-2 gene expression by RT-PCR, but also for detection of bax and bcl 2 protein expression and distribution by immunohistochemical analysis.RESULTS: The rat survival rates in aFGF treated group were higher than group R (P＜0.05) and the improvement of intestinal histological structures was observed at 2, 6, and 12 h after the reperfusion in group A compared with group R. The apoptotic rates were (41.17±3.49)%, (42.83±5.23)% and (53.33±6.92)% at 2, 6 and 12 h after reperfusion, respectively in group A, apparently less than those of group R at matched time points (50.67±6.95, 54.17±7.86, 64.33±6.47, respectively) (P＜0.05). The bax gene transcription and translation were significantly decreased in group A vs group R, while mRNA and protein contents of Bcl-2 in group A were obviously higher than those in groupR during 2-12 h period after reperfusion.CONCLUSION: The changes in histological structure and the increment of apoptotic rate indicated that the intestinal barrier was damaged after intestinal I/R injury, whilst intravenous aFGF could alleviate apoptosis induced by ischemia and reperfusion in rat intestinal tissues, in which genes of bax and bcl-2 might play important roles.