Prognostic Significance of p53 and Ki67 Proteins Expression in Greek Gastric Cancer Patients

Aim: The variability of prognosis of gastric cancer (GC) within a pathological stage necessitates the identification of subgroups of patients with a more aggressive disease. The role of p53 and Ki67 expression in gastric carcinoma is far from being fully established. The aim of the present study was to evaluate the expression of p53 and Ki67 in gastric cancer and correlate the findings with several clinicopathological features and prognosis. Materials and methods: Tissue samples from 93 patients treated by gastric resection for gastric carcinoma between 1996 and 2001 were used. Formalin-fixed paraffin-embedded tumors were studied by immunohistochemistry, using monoclonal antibodies to p53 and Ki67. The results were correlated with clinicopathological features and survival. Results: Stronger expression of p53 was related with tumor size greater than 5 cm and advanced stage. Stronger expression of Ki67 correlated with higher ratio of the number of metastatic lymph nodes to the total number of dissected lymph nodes (metastatic lymph node [MLN] ratio) and advanced stage. Moreover, p53 and Ki67 overexpression, tumor size greater than 5 cm, MLN ratio, depth of invasion, lymph node metastasis, stage III and IV and infiltrative macroscopic appearance were adverse prognostic factors. The levels of p53 and Ki67, the MLN ratio, the tumor size (above 5 cm) and the stage of the disease were identified as independent prognostic factors of survival.

Conclusions: In gastric cancer, the expression of p53 and Ki67 provides significant information about prognosis. The routine evaluation of p53 and Ki67 levels could be a useful tool in identification of patient with more aggressive disease and contribute to a better therapeutic approach.     

养胃抗瘤冲剂对胃癌患者外周血T淋巴细胞rDNA的转录活性和凋亡相关蛋白影响的研究

目的 :观察养胃抗瘤冲剂对胃癌患者外周血T淋巴细胞rDNA转录活性和凋亡相关蛋白变化的影响及在肿瘤转移中的意义。 方法 :采用KL型肿瘤免疫图像分析系统和流式细胞仪测定正常人和胃癌患者养胃抗瘤冲剂治疗前后外周血T淋巴细胞rDNA的转录活性和调亡相关蛋白 (Fas/FasL)的表达变化。 结果 :胃癌患者外周血T淋巴细胞rDNA转录活性明显降低 ,凋亡相关蛋白Fas表达升高 ,与健康人相比 ,差异显著 (P <0 0 1) ,随着病情的进展 ,这种趋势更为明显。养胃抗瘤冲剂治疗后外周血T淋巴细胞的Ag -NORs升高、Fas表达降低 ,与治疗前比较差异显著 (P <0 0 1)。 结论 :养胃抗瘤冲剂可以降低外周血T淋巴细胞的凋亡能力 ,对增殖能力则有增强作用 ,从而提高机体的细胞免疫功能。外周血T淋巴细胞rDNA转录活性和凋亡相关蛋白变化可以从淋巴细胞增殖和凋亡活性两方面评价肿瘤患者的免疫状态 ,在胃癌转移的临床监测中具有重要意义。     

体外模拟气腹环境对胃癌细胞增殖凋亡的影响

目的通过体外模拟气腹环境,观察二氧化碳(CO2)和氦气(He)在不同压力下对胃癌细胞MKN-45增殖、凋亡的影响。方法实验分为对照组、CO2组和He组,其中CO2组和He组又按照压力不同分为0、5、10、15mmHg组(作用时间均为4h)。细胞在密闭培养箱内经不同压力CO2或He作用4h后用血气分析仪检测培养液pH值;MTT法检测细胞增殖情况;流式细胞仪观察细胞凋亡指数变化;AnnexinV-FITC/PI双标染色、流式细胞仪测定凋亡细胞比例。结果处理结束后即刻检测,CO2组细胞培养液呈弱酸性,He组细胞培养液呈弱碱性。MTT法检测细胞增殖变化,CO2组0、5、10mmHg组OD值与对照组比较无明显差异(P>0.05),15mmHg组OD值与对照组比较在前3天无明显差异(P>0.05),在4~7天OD值明显低于对照组(P<0.01)。而在He组,不同压力组细胞增殖活性与对照组比较无明显差异(P>0.05)。在CO2环境下,0、5mmHg组细胞凋亡指数与对照组比较无明显差异(P>0.05),而10、15mmHg组明显大于正常对照组(P<0.01)。0、5mmHg组细胞凋亡比例与对照组比较无显著差异(P>0.05...     

Examination of soluble Fas (sFas) and soluble Fas ligand (sFasL) in patients with burns

The FasL–Fas system is one of the recognized apoptosis-inducing systems, and has been determined to have important functions in relation to homeostasis and biological defense mechanisms. In this study, we investigated the serum levels of soluble Fas (sFas), soluble FasL (sFasL) and tumor necrosis factor (TNF-) in patients with burns. The sFas levels were found to be significantly higher in the patients who eventually died as compared to those in the patients who survived (3.9±1.8 ng/ml versus 2.6±1.0 ng/ml). On the other hand, the sFasL levels were significantly higher in the patients who survived (61.5±29.9 ng/ml versus 37.2±14.4 ng/ml) than in those who eventually died. A positive correlation was noted between the TNF- level and the sFas level, and a negative correlation was observed between the TNF- level and the sFasL level. These findings suggest that worsening of the condition of a burns patient may be related to changes in the Fas–FasL system.     

Prognostic Significance of Level and Number of Lymph Node Metastases in Patients with Gastric Cancer

BACKGROUND: To present data that provide some insight into the appropriateness of a nodal grouping category and its relation to survival in patients with gastric cancer. METHODS: We reviewed data of 777 patients with advanced gastric cancer who had undergone curative gastrectomy to investigate the prognostic significance of level and number of lymph node metastases. RESULTS: The prognosis of patients with gastric cancer was well correlated with the level and number of lymph node metastases. Multivariate analysis indicated that the level and number of lymph node metastases were independent prognostic indicators. Moreover, the number of lymph node metastases was an independent prognostic factor in N1, N2, and N3 patients. The most statistically significant difference in disease-specific survival was observed at a threshold of 11 lymph node metastases, yielding a chi2 value of 42.88, a hazard ratio of 2.523, at a 95% confidence interval of 1.913, 3.329 (P < .0001) by Cox proportional hazard model. On the basis of this result, patients were divided into two groups as follows: marked lymph node metastasis group (number of positive nodes > or =11) and slight lymph node metastasis group (number of positive nodes < or =10). The prognosis of patients with marked lymph node metastasis was statistically significantly worse than that with slight lymph node metastasis in N1, N2, and N3 patients. CONCLUSIONS: Both level and number were indispensable for evaluating lymph node metastasis. Therefore, addition of the number of positive nodes to the N category defined by the Japanese Classification of Gastric Carcinoma may be a useful strategy in the N staging classification in gastric cancer.     

TRAIL信号通路在胃癌的研究现状

目的介绍肿瘤坏死因子相关凋亡诱导配体（TRAIL）及其受体在胃癌的研究现状。方法检索PUB-MEDLINE和中国期刊全文数据库（CJFD），综述国内、外近年关于TRAII．及其受体在胃癌的相关研究文献。结果TRAIL在胃癌组织表达水平报道情况差异较大，但其与胃癌的分化程度和浸润、转移情况密切相关。其受体DR4及DR5在胃癌组织均表达阳性，而DcR1及DcR2在胃癌组织亦有表达阳性的报道。caspase-3、caspase-8和survivin对胃癌TRAIL信号通路有重要调节作用。5-氮2，-杂脱氧胞苷、阿霉素、5-氟尿嘧啶、α-生育酚及X射线照射可协同增强TRAIL对胃癌细胞的凋亡诱导作用。结论胃癌可能适合TRAIL靶向治疗，但其作用机理较为复杂并受到多因素影响。尚有如何有效增强和调控TRAIL凋亡诱导作用及TRAIL有何潜在毒性等诸多问题亟待研究。     

Prognostic Significance of the Cytologic Features of Free Cancer Cells in the Peritoneal Cavity of Patients with Gastric Cancer

The aim of this study was to determine whether the cytologic features of cancer cells in the peritoneal lavage fluid from patients undergoing surgery for gastric cancer affect prognosis. A total of 141 patients with stage III–IV gastric cancer were enrolled in this study. Among 111 patients without peritoneal metastasis (P0), 43 were noted to have positive peritoneal cytology (P0/CY1). These patients were subclassified into two groups according to the cluster formation of cancer cells in the cytology specimen, namely, 17 with cluster formation and 26 without cluster formation. The clinicopathologic parameters and survival rates were compared between these two groups. Among the P0/CY1 patients, no significant difference in clinicopathologic features existed between patients with or without the cluster formation of cancer cells. However, the survival rate of patients without cluster formation was significantly higher than that of those with cluster formation. Furthermore, the survival curves of patients without cluster formation was similar to that of the P0/CY0 patients with stage IV cancer. Subclassification according to the presence or absence of the cluster formation of cancer cells may be useful for predicting the outcome of patients with positive cytology after surgery for advanced gastric cancer. Received: March 9, 2001 / Accepted: July 17, 2001     

Expression and Prognostic Significance of Pepsinogen C in Gastric Carcinoma

Background: In this study we evaluated the expression and clinical significance of pepsinogen C, an aspartic proteinase involved in the digestion of proteins in the stomach, in patients with gastric cancer.Methods: Pepsinogen C expression was examined by immunohistochemical methods in a series of 95 gastric carcinomas. The prognostic value of pepsinogen C was retrospectively evaluated by multivariate analysis taking into account conventional prognostic parameters. Follow-up period of patients was 21.4 months.Results: A total of 25 (26.3%) gastric carcinomas stained positively for pepsinogen C. The percentage of pepsinogen C-positive tumors was higher in well-differentiated (50%) than in moderately differentiated (19.5%) and poorly differentiated (21.9%) tumors (P < .05). Similarly, significant differences in pepsinogen C immunostaining were found between node-negative and node-positive tumors (47.1% vs. 14.7%; P < .001). In addition, statistical analysis revealed that pepsinogen C expression was associated with clinical outcome in gastric cancer patients. Low pepsinogen C levels predicted short overall survival periods in the overall group of patients with gastric cancer (P < .001), and in 71 patients with resectable carcinomas (P < .005). Multivariate analysis according to Coxs model indicated that pepsinogen C immunostaining was an independent predictor of outcome for both overall and resectable gastric cancer patients (P < .05, for both).Conclusions: The expression of pepsinogen C in gastric cancer may represent a useful biological marker able to identify subgroups of patients with different clinical outcomes.     

抑癌基因Lumican在胃癌中的表达及其临床意义

目的探讨抑癌基因Lumican mRNA在胃癌组织中的表达及其在胃癌发生、发展中的意义。方法采用逆转录聚合酶链反应(RT-PCR)方法检测66例胃癌组织及其相应的近癌旁组织及正常组织中LumicanmRNA的表达,并分析胃癌组织中Lumican mRNA表达与其临床病理特征的关系。结果Lumican mRNA在胃癌组织、近癌旁组织及正常组织中的表达缺失率分别为42.4%(28/66)、15.2%(10/66)及0(0/66)。有淋巴结转移者胃癌组织中Lumican mRNA表达缺失率(61.1%)明显高于无淋巴结转移者(20.0%),差异有统计学意义(χ2=11.323,P=0.001);晚期(Ⅲ、Ⅳ期)胃癌组织中Lumican mRNA表达缺失率(61.5%)明显高于早期(Ⅰ、Ⅱ期,30.0%),差异有统计学意义(χ2=6.417,P=0.011;胃癌组织中Lumican mRNA表达缺失与肿瘤分化程度无关(χ2=1.576,P=0.455)。结论Lumican mRNA表达缺失可能在胃癌的发生和发展过程中起重要作用,并影响其预后。     

Fas/FasL表达和细胞凋亡在人类同种异体移植肾急性排斥中的作用

目的探讨移植肾急性排斥（ＡＲ）时细胞凋亡与Ｆａｓ和Ｆａｓ配体（ＦａｓＬ）表达的作用及其临床意义。方法分别用原位末端标记技术（ＴＵＮＥＬ法）和免疫组织化学方法检测２６例移植肾ＡＲ标本中细胞凋亡和Ｆａｓ／ＦａｓＬ表达情况。结果细胞凋亡和Ｆａｓ／ＦａｓＬ表达主要在ＡＲ移植肾小管上皮发生,且凋亡指数和Ｆａｓ／ＦａｓＬ表达与肾组织病理损伤程度平行,与正常肾对照组和移植肾功能稳定组比较差异显著（Ｐ＜０．０１）。结论肾小管上皮细胞凋亡在ＡＲ所致的移植肾损伤中起重要作用,Ｆａｓ／ＦａｓＬ系统可能参与移植肾ＡＲ,是造成肾小管上皮细胞凋亡的重要因素。ＴＵＮＥＬ法检测细胞凋亡可作为判断移植肾病理变化和预后的重要指标     

狼疮肾炎患者外周血单核细胞表达有功能的Fas

目的 探讨狼疮肾炎（ＬＮ）患者单核细胞凋亡加速的机制。方法 对灯例ＬＮ患者进行观察及以 １５例正常人为对照。采用流式细胞仪检测单核细胞 ＦａＳ和 Ｆａｓ Ｌｉｇａｎｄ（ＦａｓＬ）的表达；单核细胞与重组人ＦａＳＬ共孵育，碘化丙啶染色、流式细胞仪检测单核细胞凋亡，四甲基偶氮唑蓝（ＭＴＴ）法观察单核细胞体外存活率。结果ＬＮ患者单核细胞Ｆａｓ的表达较正常人明显上调（Ｐ＜０．０５）；处于狼疮活动期的ＬＮ患者与狼疮静止期的ＬＮ患者之间Ｆａｓ的表达差异无显著性意义（Ｐ＞０．０５）。正常人和ＬＮ患者的单核细胞表面均未检测出ＦａｓＬ的表达。单核细胞与重组人ＦａｓＬ在体外共同培养２ｈ后，ＬＮ者单核细胞的凋亡较正常人明显增加（Ｐ＜０．０５），单核细胞与重组人 ＦａｓＬ在体外共同培养６ｈ后，ＬＮ患者单核细胞的存活率较正常人明显低下（Ｐ＜０．０５）。结论 ＬＮ患者单核细胞表面功能性Ｆａｓ表达上调是ＬＮ患者单核细胞凋亡加速的可能机制。     

冠状动脉搭桥术后血浆可溶性Fas及其配体的变化

目的比较在心肺转流（CPB）或非CPB下冠状动脉搭桥术（cABG）后可溶性Fas（sFas）和可溶性Fas配体（sFasL）的血浆浓度变化。方法19例病人分别在CPB（CPB组，n=9）或非CPB（非CPB组，n=10）下行择期cABG。术前、术毕和术后取血测定白细胞介素-6（IL-6）、中性粒细胞弹性蛋白酶、sFas、sFasL血浆浓度。结果术毕、术后4h的IL-6和术毕、术后4、12h的中性粒细胞弹性蛋白酶非CPB组明显低于CPB组（P〈0．05或P〈0．01）。CPB组的sFas在术后4、12h明显升高（P〈0．05或P〈0．01），非CPB组在术后12h明显升高（P〈0．01），术后24h恢复至术前水平。两组的sFasL在术毕和术后4、12h均明显升高（P〈0．05或P〈0．01），术后24h恢复至术前水平；其中术后12h非CPB组明显低于CPB组（P〈0．05）。结论CPB或非CPB下行cABG均导致sFas和sFasL血浆浓度升高，但CPB的应用使sFasL血浆浓度升高得更多。sFasL血浆浓度可反映机体炎性反应的程度。     

胃癌术中盆腹膜取材检测微转移及其临床意义评估

目的采用胃癌术中直肠子宫（膀胱）陷窝腹膜活检并应用CEA、CK-20免疫组织化学染色的方法确定胃癌腹膜远处的微转移。方法2004年6月至2006年3月期间，选择南京市鼓楼医院40例术前物理学检查和影像学检查未能发现，术中无肉眼及用手可触及的腹膜远处转移的胃癌病例，行术中直肠子宫（膀胱）陷窝腹膜活检并行HE常规染色和CEA、CK-20免疫组织化学染色检查有无腹膜远处微转移。结果本组40例胃癌患者中有10例发生直肠子宫（膀胱）陷窝腹膜的微转移，均发生在肿瘤侵及胃壁全层或浆膜外者[27．8％（10／36）]，其微转移发生率明显高于肿瘤未侵及胃壁全层或浆膜外者[0（0／4）]，P〈0．05。10例中8例淋巴结转移数目均超过6枚，另外2例淋巴结转移数目均为1枚；转移淋巴结数≥7枚时的腹膜微转移发生率[44．4％（8／18）]明显高于转移淋巴结数〈7枚者[16．7％（2／12）]，P〈0．05。10例中，有8例为低分化腺癌，2例为中分化腺癌。结论侵及浆膜或浆膜外的胃癌病例，即使在术前检查或术中探查时未发现远处腹膜转移，术中仍可行直肠子宫（膀胱）陷窝腹膜活检，以发现有无远处腹膜的微转移，为胃癌分期、腹腔内辅助化疗及预测预后提供证据。     

The Immunohistochemical Localization of Fas and Fas Ligand in Jaw Bone and Tooth Germ of Human Fetuses

The cellular localization and roles of bone morphogenetic protein (BMP)-2 and apoptosis-associating factors in human orofacial development remain unclear. In this study, BMP-2, osteocalcin, and TGF-β, which are bone-differentiating markers, apoptosis-associating factors (i.e., Bcl-2, Bax, Fas, and Fas ligand), apoptotic cells detected by the in situ 3′-end labeling method (TUNEL), and proliferating cell nuclear antigen (PCNA) were immunohistochemically examined in the heads (in particular, the jaw bone and tooth germs) of human fetuses of 11-week pregnancy. BMP-2 was positive in osteoblasts and newly formed osteoid of the incisive and palatal bone of the maxilla and the mandible, which indicated that BMP-2 was exclusively involved in intramembranous ossification in the human fetal head. Fas was positive in the cytoplasm of osteocytes and a few osteoblasts. In contrast, Fas ligand was positive in the cytoplasm of osteoblasts and abundant in the stroma of the osteoblastic layer, periosteum, and perichondrium. The Fas ligand in the stroma was recognized as the soluble form, which was possibly produced by osteoblasts. TUNEL-positive apoptotic cells were found in a few osteocytes and a few osteoblastic cells in new bone, and in monocytes of degenerate Meckel's cartilage. The induction of apoptosis observed in monocytes seems to be caused via a Fas-Fas ligand cell death system, because some of these monocytes were Fas-positive, and most of them were Fas ligand-positive. Interestingly, the abundant soluble Fas ligand observed in the periosteum probably protects the bone-formative zone from the invasion of the activated lymphocytes by binding to Fas expressing in these lymphocytes and killing these cells. Fas and Fas ligand were focally positive in the dental lamina and inner enamel epithelium and cusps of the enamel organ, nevertheless, the presence of TUNEL-positive cells was very rare. Bcl-2 was clearly and Bax was weakly positive in the cells throughout the dental lamina and enamel organ. These findings indicated that Fas-mediated apoptosis was inhibited by the Bcl-2 family in the development of teeth. Received: 22 January 1998 / Accepted: 2 December 1999     

术前胃动脉化疗栓塞对胃癌淋巴结转移灶癌细胞凋亡的影响

目的探讨术前胃动脉化疗栓塞对胃癌淋巴结转移灶癌细胞凋亡的影响。方法对40例胃癌伴淋巴结转移患者行胃癌根治性切除术，根据术前是否行胃动脉化疗栓塞分为治疗组和对照组，各20例，采用免疫组化ABC法检测淋巴转移灶p53、bcl-2及CD95基因的表达，原位末端脱氧核苷转移酶标记法（TUNEL）检测细胞凋亡情况。结果治疗组淋巴转移灶p53和CD95表达较对照组明显增强（P〈0．05），并与细胞凋亡呈正相关，bcl-2表达则减低与细胞凋亡呈负相关。结论胃癌术前行胃动脉化疗栓塞，可能通过p53、bcl-2及CD95介导使转移淋巴结癌细胞凋亡增加，有助于提高根治性手术的治疗效果。     

胃动素在胃癌组织中的表达及其意义

目的检测胃动素在胃癌组织中的表达并探讨其与胃癌临床病理特征的关系。方法采用免疫组化SP法检测胃癌、癌旁及正常胃黏膜组织中胃动素蛋白的表达,对胃癌组织中胃动素蛋白的表达及其与胃癌临床病理特征之间的关系进行分析。结果胃癌组织中胃动素蛋白表达水平(1 206.43±631.67)明显高于正常胃黏膜组织(890.55±98.67)和癌旁组织(922.75±265.99),P<0.01,而癌旁组织和正常胃黏膜组织中胃动素蛋白表达水平的差异无统计学意义(P>0.05)。胃动素蛋白表达与肿瘤部位、肿瘤分化程度及淋巴结转移有关(P<0.05)。结论胃动素可能参与胃癌的发生,并与胃癌的侵袭和转移相关。     

A biochemical mechanism for resistance of intervertebral discs to metastatic cancer: Fas ligand produced by disc cells induces apoptotic cell death of cancer cells

Metastatic spinal cancer is characterized by the maintenance of normal disc structure until the vertebral body is severely destroyed by cancer cells. Anatomic features of the discs have been thought to be the main factor which confer the discs their resistance to metastatic cancer. However, little is known about the biochemical mechanism to prevent or attenuate the local infiltration of cancer cells into the discs. The purpose of this study was to investigate whether Fas ligand (FasL) produced by disc cells can kill Fas-bearing breast cancer cells by Fas and FasL interaction. Two human breast cancer cells (MCF-7 and MDA-MB-231) were obtained and cultured (1 × 10⁶ cells/well), and the expression of Fas was investigated by western blot analysis. Annulus fibrosus cells were isolated and cultured, and the presence of FasL was quantified in the supernatants of three different numbers of annulus fibrosus cells (1×, 2×, and 4 × 10⁶ cells/well) by ELISA assay. The MCF-7 and MDA-MB-231 cancer cells were cultured with supernatants of annulus fibrosus cells for 48 h. As controls, MCF-7 and MDA-MB-231 cancer cells were also cultured by themselves for 48 h. Finally, we determined and quantified the apoptosis rates of MCF-7 and MDA-MB-231 cancer cells by Annexin V–FITC and PI and TUNEL at 48 h, respectively. The expression of Fas was identified in MCF-7 and MDA-MB-231 cancer cells. The mean concentrations of FasL in supernatants of annulus fibrosus cells (1×, 2×, and 4 × 10⁶ cells/well) were 10.8, 29.6, and 56.4 pg/mL, respectively. After treatment with the supernatant of three different numbers of annulus fibrosus cells, the mean apoptosis rate of MCF-7 cancer cells was increased (2.8%, P < 0.01; 6.7%, P < 0.001; 31.0%, P < 0.001) in a dose-dependent manner of FasL compared to that of control (1.1%). The mean apoptosis rate of MDA-MB-231 cancer cells was also increased (5.7%, P < 0.01; 11.1%, P < 0.001; 25.3%, P < 0.001) in a dose-dependent manner of FasL compared to that of control (2.1%). TUNEL also demonstrated direct evidence of apoptoses of MCF-7 and MDA-MB-231 cancer cells. Our results demonstrate that Fas-bearing cancer cells undergo apoptosis by FasL produced by disc cells, which may be considered as a potential biochemical explanation for the disc’s resistance to metastatic cancer. This is the EuroSpine Winning Paper 2007 on Basic Science.     

Bcl基因与Ki—67抗原在胃癌组织中表达的研究

目的:探讨bcl-2基因和Ki-67抗原在胃癌中的表达及其可能的作用和意义.方法:用免疫组织化学LSAB法检测bCI-2和Ki-67在胃癌中的表达.结果:正常胃粘膜中bcl-2基本无表达.慢性萎缩性胃粘膜中可见基底部有bcl-2表达.7例胃癌bcl-2表达阳性,其中肠型胃癌阳性高于弥漫型.bcl-2的表达与胃癌的浸润深度和淋巴结转移无关.弥漫型胃癌的Ki-67表达显著高于肠型胃癌.Ki-67指数随浸润深度的增加而增高.有淋巴结转移的Ki-67指数显著高于无转移者.结论:bcl-2的表达可能对胃癌特别是肠型胃癌的发生有重要作用.Ki-67的表达与胃癌的发展及其预后的判断可能有意义.