Correlation between survivin expression and prognosis in non-small cell lung cancer

AIMS AND BACKGROUND: Survivin is a recently identified protein as an inhibitor of apoptosis, which supresses programmed cell death and regulates cell division. In this study, we investigated the prognostic significance of both nuclear and cytoplasmic survivin expression in non-small cell lung cancer (NSCLC) and examined the association with clinicopathological parameters. METHODS: The study comprised 58 male patients diagnosed NSCLC with a mean age of 57.29+/-8.82 years; range 40-76 years. Patients underwent lobectomy (64%) or pneumonectomy (36%) with hilar and mediastinal lymph node sampling. Paraffin embedded tumor sections were retrieved for evaluation of nuclear and cytoplasmic staining of survivin. Clinicopathological data, stage and survival of patients were all determined. RESULTS: Cytoplasmic staining was found significantly increased in squamous cell carcinoma (P=0.003), whereas there was no association between nuclear staining and histopathological type (P=0.837). Also, both nuclear and cytoplasmic staining did not show any association with tumor stage (P>0.05). In univariate analysis there was significant correlation between nuclear survivin and short survival (P=0.0002). In multivariate survival analysis using Cox regression, only nuclear staining of survivin was determined as an independent prognostic factor (P=0.004). CONCLUSIONS: Localization of survivin expression might have an important regulatory mechanism in carcinogenesis and tumor progression. Nuclear survivin expression in tumor tissues might predict the prognosis in NSCLC, whereas cytoplasmic survivin has no prognostic significance.     

肝癌细胞放射敏感性与survivin蛋白表达的关系

目的 探讨肝癌细胞放射敏感性与survivin表达的关系.方法 肝癌细胞HepG2和SMMC-7721在接受不同剂量γ射线照射后,分别采用克隆形成法、免疫细胞化学法、流式细胞术、比色法等检测细胞存活率、survivin蛋白表达、细胞周期变化和Caspase-3活性.结果 在2Gy照射下HepG2和SMMC-7721细胞的存活分数分别为0.43±0.01与0.70±0.02,SMMC-7721较HepG2放射抗拒.γ射线对SMMC-7721细胞的G2/M期阻滞时间较HepG2细胞长(48 h对24 h),在阻滞峰各剂量点SMMC-7721细胞的G2/M期比例也更高.γ射线可上调两株肝癌细胞survivin蛋白的表达,照射后48～72 h,SMMC-7721细胞的survivin蛋白表达水平显著高于HepG2细胞(t值为2.81～5.20,P值均＜0.05).而Caspase-3的活化水平在放射敏感的HepG2细胞中更高(t值为6.05～6.72,P值均＜0.01).结论 射线诱导的survivin表达上调及survivin对Caspase-3的负调控可能是SMMC-7721细胞较HepG2细胞放射抗拒的原因之一.     

Relationship of CD15 immunoreactivity and prognosis in sporadic medullary thyroid carcinoma

Summary Immunoreactivity with monoclonal antibody CD15 (Leu-M1) was investigated in the primary tumours, the metastases and local recurrences of 47 cases of sporadic medullary carcinoma of the thyroid (MTC). Of these tumours, 36.5% showed a varying degree of CD15 immunostaining; in 7 carcinomas the CD15 immunoreactivity was found to be significant (>15% tumour cells positively stained). Staining of the amyloid stroma was observed in 3 tumours. Significantly higher epithelial CD15 positivity was seen more frequently in the group with larger tumours (>4 cm) and was found exclusively in the presence of lymph node metastases. No substantial difference in the percentage of immunostained cells was seen between primary tumours and metastatic or recurrent lesions, except for two cases that revealed a significant increase in the number of CD15-immunostained cells in metastatic and recurrent lesions. Five of 7 patients with recurrences showing significant CD15 immunostaining died of cancer, while in the absence of significant CD15 staining all patients with recurrences were still alive at the conclusion of the study. The prognostic value of CD15 immunoreactivity, found by univariate analysis, becomes weaker after adjustment for the size and stage of tumour. Particularly in patients with tumour recurrences CD15 immunostaining may be of clinical relevance for the selection of patients in whom a more radical surgical approach would be justified.Abbreviations MTC medullary thyroid carcinoma     

Survivin与肝癌关系的研究进展

Survivin是一种凋亡抑制蛋白,具有调节细胞分化和抑制凋亡的双重功能.因其在人类大多数肿瘤中高表达,故成为近年来肿瘤基因治疗研究热点.此文就Survivin在肝癌中的研究进展作一综述.     

Relationship between expression of Smac and Survivin and apoptosis of primary hepatocellular carcinoma

Introduction Disturbance of the regulation of cell apoptosis can lead to cell over-proliferation, and decrease of apoptosis can result in tumorigenesis ortumor development. The apoptotic signaling pathway is regulated by a variety of factors and is based on the balance between cell death and survival factors.[1, 2] The central players in apoptosis are caspases, and one important route to activation of the caspases involves the translocation of cytochrome C (Cyt-C) from mitochondria to cytosol…     

生存素基因在胃癌组织中的表达及其与胃癌预后关系

目的探讨生存素(survivin)基因在胃癌组织中的表达及其与胃癌预后关系。方法病例来源于1995年7月至1996年6月中山医院住院手术患者，采用免疫组化Envision二步法检测手术切除的胃癌病灶组织中生存素基因表达。根据手术病理诊断与临床表现确定肿瘤TNM分期。所有病例随访至少5年或直到死亡。比较不同病理类型、不同TNM分期生存素表达差异。按生存素基因是否表达比较两组患者生存曲线的差异。结果共有96例患者进入研究和随访观察，其中男59例，女37例；年龄29～84岁，平均56岁，68例表达生存素基因产物，阳性率70．8％。病理类型中腺癌78例，非腺癌18例，不同病理类型生存素基因表达相似(腺癌69．7％、非腺癌60．0％，P=0．369)。对腺癌病例分析显示：低分化组生存素基因表达高于中高分化组(82．1％比62．5％，P=0．053)；肿瘤累及固有肌层组和全层组生存素基因表达高于肿瘤局限于黏膜或黏膜下层组(81．3％，75．9％比33．3％，P=0．020)。生存素表达与淋巴结转移与否无关(68．1％比70．8％，P=0．771)。生存素阳性组5年生存率(42．93％)低于生存素阴性组(52．93％)，但差异无统计学意义。结论生存素基因在胃癌中高表达。生存素基因表达与胃腺癌分化程度以及肿瘤累及深度有关，需进行更多研究评价其在预后中的作用。     

Glypican-3与原发性肝癌的关系研究进展

近年来有关Glypican-3（GPC3）与原发性肝癌（HCC）关系的研究逐年增多,发现GPC3无论在原发性肝癌病人的血清中还是肿瘤组织中均有较高的表达,对原发性肝癌的诊断具有较高的灵敏性和特异性。其表达水平与原发性肝癌的预后也有一定的关系。同时,GPC3也为原发性肝癌的治疗带来了新的思路。     

A comparative study of the expression of Wnt-1, WISP-1, survivin and cyclin-D1 in colorectal carcinoma

Background and aims It is well accepted that activation of Wnt signalling occurs in colorectal carcinoma (CRC), but the correlation amongst the various proteins involved in primary tumours are still unclear. The expression of the inducer of this pathway, Wnt-1, and the downstream effectors, WISP-1, cyclin-D1 and survivin proteins, was compared in a series of CRC tissues with the apparently normal adjacent tissues to determine the relationship of these proteins.Patients and methods Formalin-fixed, paraffin-embedded tissue samples of 47 CRCs surgically resected at the Kuala Lumpur Hospital (KLH) between 1999 and 2000 were used. Immunohistochemical staining with monoclonal antibodies against cyclin-D1 and survivin and polyclonal antibodies against Wnt-1 and WISP-1 was performed. Results of immunohistochemistry were analysed for correlation between biomolecules and histopathological data of the patients.Results Of the 47 CRCs, 26 (55.3%), 15 (31.9%), 5 (10.6%) and 28 (59.6%) of the tumours exhibited positivity for Wnt-1, WISP-1, cyclin D1 and survivin, respectively. A lower percentage of the 40 apparently normal adjacent tissues were found to be positive for Wnt-1 (7, 17.5%), WISP-1 (±5, 12.5%) and survivin (13, 32.5%), but cyclin D1 was not detected in any of them. Interestingly, the total scores of Wnt-1, WISP-1 and survivin were significantly higher in CRC tissues (p=0.001, 0.034 and 0.044, respectively). Using the Spearman rank correlation test, a positive linear relationship was found between total Wnt-1 score with total WISP-1 score (rho=0.319, p=0.003) and total survivin score (rho=0.609, p=<0.001). The expression of WISP-1 in the CRC tissues was found to be positively correlated with patients older than 60 years old (p=0.011). In addition, nuclear cyclin-D1 expression was found to be associated with poorly differentiated CRC tissues (p<0.001, Table 5) and right-sided CRC tumour (p=0.019, Table 6). Total WISP-1 score was associated with well-differentiated CRC tissues (p=0.029).Conclusions Overexpression and interplay between Wnt-1, WISP-1, survivin and cyclin-D1 may play a role in tumorigenesis, possibly by promoting cell cycle checkpoint progression, accelerating cell growth and inhibiting apoptosis. Our data may provide useful information towards the search for potent therapeutic targets towards the development of novel treatment strategies for CRC.     

人肝癌多药耐药细胞株survivin蛋白的表达及意义

目的探讨人肝癌多药耐药细胞株survivin蛋白表达与肝癌细胞多药耐药的关系.方法用阿霉素(DOX)浓度梯度递增导法,建立人肝癌多药耐药细胞株(Bel-7402/DOX).用流式细胞仪检测肝癌细胞凋亡率,同时用蛋白印迹(Western blot)检测其survivin蛋白表达.结果随着培养液中 DOX 浓度的增加Bel-7402/DOX 的凋亡率逐渐增加,survivin蛋白的表达逐渐增强.结论Survivin蛋白的表达变化可能与人肝癌细胞获得性耐药有关.     

WEE1与肝癌的关系

目的:探讨WEE1在肝细胞癌(HCC)发生、发展中的异常表达及与肿瘤病理分期的关系.方法:收集2010-03/2010-05河南省人民医院肝胆外科手术标本正常肝组织23例,肝硬化20例,HCC42例.采用RT-PCR检测mRNA水平的表达,Western blot及免疫组织化学检测蛋白的表达,并分析其与肝癌临床病理学分期的关系.结果:WEE1mRNA阳性表达率分别为21.7%、55.0%和90.5%,三组间差异有统计学意义(P<0.01).Western blot和免疫组织化学方法分别检测蛋白阳性表达率分别为13.04%/17.4%、40%/60%和78.6%/83.3%,三组间相比差异有统计学意义(P<0.01).肝癌组织中WEE1强度与肿瘤的分化程度及病理分级相关(χ2=17.454,P<0.01;χ2=14.559,P<0.01).结论:WEE1基因在肝癌中呈上调表达,其参与的DNA的修复异常与肝癌的发生密切相关; 且与肿瘤的分化程度和病理分级相关.     

Expression and significance of new inhibitor of apoptosis protein survivin in hepatocellular carcinoma

AM: To investigate expression and significance of inhibitor of apoptosis protein survivin in hepatocellular carcinoma (HCC). METHODS: The expression of survivin and vascular endothelial growth factor (VEGF) was investigated in 38 cases of HCC tissues and 38 liver cirrhosis tissues by immunohistochemistry and Western blot. The relationship between the expression of survivin and clinicopathological factors of HCC was analyzed. RESULTS: Survivin protein was detected in 23 (60.5%) of 38 HCCs and 3 (7.9%) of 38 liver cirrhosis tissues. In 23 cases of HCC which expressed survivin, the expression of VEGF was positive in 18 cases and slight positive or negative in 5 cases. While in 15 cases of HCC which did not express survivin, 12 cases did not express or slightly expressed, and 3 cases expressed VEGF. In liver cirrhosis tissues, the expression of VEGF was as follows: 24 cases were negative, 10 cases were weak positive and 4 cases were strong positive. The expression of survivin was coincident with the expression of VEGF in HCC (P<0.01). The expression of survivin in HCC had no relationship with the patients' age, gender, tumor size and differentiation level of HCC, while it was related to the metastasis of HCC. The protein quantitative analysis by Western blot also showed that overexpression of survivin in HCC was closely correlated to the expression of VEGF (P<0.01). Furthermore, stronger expression of survivin and VEGF was also found in patients with metastasis rather than in those with no metastasis (P<0.01). CONCLUSION: Survivin plays a pivotal role in the metastasis of HCC, and it has some correlation with tumorigenesis. The expression of survivin in the primary lesion is very useful as an indicator for metastasis and prognosis of HCC. It could become a new target of gene therapy of HCC.     

EZH2和VEGF在结直肠癌中的表达其与患者预后关系生物信学分析及验证

背景结直肠癌(colorectal carcinoma, CRC)是临床上常见的消化系统恶性肿瘤.但其确切发病机制和预后独立因素仍未阐明.本研究通过生物信息学方法分析了zeste基因增强子同源物2(enhancerofzestehomolog2,EZH2)和血管内皮生长因子(vascular endothelial growth factor, VEGF)基因在CRC中的表达情况及其和患者预后的感谢.并采用免疫组化检测了EZHE2和VEGF蛋白的表达情况及其与患者临床特征的关系.目的探讨EZH2和VEGF在CRC中的表达、突变情况及其与患者临床病理特征和预后的关系.方法首先在TCGA数据库中比较EZH2和VEGF基因mRNA在肠癌患者癌和癌旁组织中的表达,同时分析EZH2和VEGF基因的突变情况;采用STRING数据库建立EZH2和VEGF基因表达网络并筛选网络中的关键基因.根据EZH2和VEGF在肿瘤组织中的表达分为高低表达组, Cox回归模型Log-rank检验比较高低表达组患者总生存和无疾病进展生存是否存在差异.同时选取80例肠癌手术患者,留取患者癌组织和癌旁组织,采用免疫组织化学法检测上述组织中EZH2和VEGF蛋白表达水平.结果 TCGA数据库显示EZH2和VEGF基因在CRC组织中的表达水平显著高于对应的正常肠上皮组织(P <0.05),而与肠癌患者的临床分期并无明显相关性(P>0.05);EZH2和VEGF基因在人肠癌中的突变率分别为1.5%和1.9%,且EZH2和VEGF不同突变组织中mRNA表达水平存在明显差异.网络中共有22个蛋白,各蛋白平均相互作用指数为10.5,区域聚集指数为0.8,各蛋白富集明显(P<0.01). Cytohubb软件筛选出EZH2, DNMT1, HDAC2, YY1和SUZ12为网络中的关键基因;EZH2和VEGF高低表达与患者总生存期均无相关性(P>0.05),而VEGF高表达组患者无疾病进展生存期显著低于低表达组(HR=1.8, P<0.05).免疫组化显示, EZH2和VEGF在肠癌组织中的阳性表达率均显著高于癌旁组织(P<0.01).EZH2阳性表达与肠癌肿瘤直径、分化程度和Duke分期有关(P<0.05).而VEGF阳性表达与肠癌患者分化程度和Duke分期存在相关性(P<0.05).结论 EZH2和VEGF在肠癌组织中呈现明显上调表达和突变,其高表达与肿瘤大小、分会程度和Duke分期有关,并可作为肠癌预后的潜在分子标志物.     

Significance and relationship between Yes-associated protein and survivin expression in gastric carcinoma and precancerous lesions

AIM： To analyze the differences and relevance of Yesassociated protein （YAP） and survivin, and to explore the correlation and significance of their expression in gastric carcinoma and precancerous lesions.
METHODS： The PV9000 immunohistochemical method was used to detect the expression of YAP and survivin in 98 cases of normal gastric mucosa, 58 intestinal metaplasia （IM）, 32 dysplasia and 98 gastric carcinoma.
RESULTS： The positive rates of YAP in dysplasia （37.5%） and gastric carcinoma （48.0%） were significantly higher than that in normal gastric mucosa （13.3%）, P 〈 0.01. The positive rates of survivin in IM （53.4%）, dysplasia （59.4%） and gastric carcinoma （65.3%） were significantly higher than in normal gastric mucosa （11.2%）, P 〈 0.01. Survivin expression gradually increased from 41.7% in well differentiated adenocarcinoma through 58.3% in moderately differentiated adenocarcinoma to 75.6% in poorly differentiated adenocarcinoma, with significant Rank correlation, rk = 0.279, P 〈 0.01. The positive rate of survivin in gastric carcinoma of diffused type （74.6%） was significantly higher than that in intestinal type （51.3%）, P 〈 0.05. In gastric carcinoma with lymph node metastasis （76.9%）, the positive rate of survivin was significantly higher than that in the group without lymph node metastasis （41.2%）, P 〈 0.01. In 98 cases of gastric carcinoma, the expression of YAP and of survivin were positively correlated, rk = 0.246, P 〈 0.01.
CONCLUSION： YAP may play an important role as a carcinogenic factor and may induce survivin expression. Detecting both markers together may help in early diagnosis of gastric carcinoma.     

Knockdown of survivin gene expression by RNAi induces apoptosis in human hepatocellular carcinoma cell line SMMC-7721

AIM: To investigate the survivin gene expression in human hepatocellular carcinoma cell line SMMC-7721 and the effects of survivin gene RNA interference (RNAi) on cell apoptosis and biological behaviors of SMMC-7721 cells. METHODS: Eukaryotic expression vector of survivin gene RNAi and recombinant plasmid pSuppressorNeo-survivin (pSuNeo-SW), were constructed by ligating into the vector, pSupperssorNeo (pSuNeo) digested with restriction enzymes Xba I and Sail and the designed double-chain RNAi primers. A cell model of SMMC-7721 after treatment with RNAi was prepared by transfecting SMMC-7721 cells with the lipofectin transfection method. Strept-avidin-biotin-complex (SABC) immunohistochemical staining and RT-PCR were used to detect survivin gene expressions in SMMC-7721 cells. Flow cytometry was used for the cell cycle analysis. Transmission electron microscopy was performed to determine whether RNAi induced cell apoptosis, and the method of measuring the cell growth curve was utilized to study the growth of SMMC-7721 cells before and after treatment with RNAi. RESULTS: The eukaryotic expression vector of survivin gene RNAi and pSuNeo-SW, were constructed successfully. The expression level of survivin gene in SMMC-7721 cells was observed. After the treatment of RNAi, the expression of survivin gene in SMMC-7721 cells was almost absent, apoptosis index was increased by 15.6%, and the number of cells was decreased in G2/M phase and the cell growth was inhibited. CONCLUSION: RNAi can exert a knockdown of survivin gene expression in SMMC-7721 cells, and induce apoptosis and inhibit the growth of carcinoma cells.     

Relationship between co-stimulatory molecule B7-H3 expression and gastric carcinoma histology and prognosis

AIM:To investigate the expression of co-stimulatorymolecule B7-H3 in gastric carcinoma and adenomatissue as well as normal gastric tissue and to explore therelationship between B7-H3 expression and pathologicalfeatures and prognosis of gastric carcinoma.METHODS:B7-H3 expression was detected in 102samples of human gastric carcinoma and 10 samples ofgastric adenoma and 10 samples of normal gastric tissueby immunohistochemical assay.Correlation betweenthe expression of B7-H3 and the patients'age,sex,gastric carcinoma locus,tumor size,tissue type,tumorinfiltration depth,differentiation degree,lymph nodemetastasis,and survival time was analyzed.RESULTS:B7-H3 was expressed in all gastric adenomasamples and in 58.8% samples of gastric carcinoma.B7-H3 expression in gastric carcinoma samples wasnot related with the patients'age,sex,lymph nodemetastasis,and tumor size(P>0.05),but with thesurvival time,infiltration depth of tumor and tissue type.CONCLUSION:Detection of B7-H3 expression in gastriccarcinoma tissue is beneficial to the judgment of theprognosis of gastric carcinoma patients and the choice oftreatment.     

Decreased expression of Neurensin-2 correlates with poor prognosis in hepatocellular carcinoma

AIM： To investigate the expression of Neurensin-2 （NRSN2） in hepatocellular carcinoma （HCC） and its prognostic values in predicting survival. METHODS： The expression and prognostic significance of NRSN2 in HCC was examined by performing immunohistochemical analysis using a total of 110 HCC clinical tissue samples, and Western blotting analysis to further confirm the result. RESULTS： Decreased NRSN2 expression was shown in 70.9% cases. Loss of NRSN2 expression in HCC was significantly related to tumor size （P = 0.006）. Larger tumor size was related to negative expression of NRSN2. Patients showing negative NRSN2 expression had a significantly shorter overall survival than those with positive expression （P = 0.008）. Multivariate Cox regression analysis indicated that NRSN2 expression level was an independent factor of survival （P = 0.013）. Western blotting analysis further confirmed decreased expression of NRSN2 in tumor tissues compared with non-tumorous tissues. CONCLUSION： Our study indicated that NRSIV2 could be a tumor suppressor gene for HCC and a candidate biomarker for long-term survival in HCC.     

Ki-67、VEGF、Cyclin D1的表达与肝细胞癌生物学行为的关系

目的:探讨肝细胞癌(hepatocellular carcinoma,HCC)、胆管癌、肝硬化及正常肝组织中增殖细胞核抗原(Ki-67)、血管内皮生长因子(vascular endothelial growth factor,VEGF)及周期素D1(Cyclin D1)蛋白表达的意义及其与肝癌生物学行为关系.方法:对HCC组织52例、胆管癌组织10例、肝硬化组织10例及正常肝组织10例,采用免疫组织化学S-P法检测Ki-67、VEGF及Cyclin D1蛋白.结果:Ki-67、VEGF及Cyclin D1在HCC的强阳性表达率分别为48.1%、55.8%及55.8%,均显著高于胆管癌、肝硬化及正常肝组织(?2=15.672、15.524、14.812,P<0.001).Ki-67与肿瘤大小、血管侵犯、分化程度有关;VEGF、Cyclin D1表达与肿瘤包膜完整、血管侵犯及肿瘤分化程度明显有关;Ki-67与VEGF及Cyclin D1间无相关.VEGF与Cyclin D1间的蛋白表达呈正相关(r=0.374,P<0.01).结论:Ki-67、VEGF及Cyclin D1表达与肝细胞癌生物学行为密切相关,与肝癌的发生和发展密切相关.关键词:肝细胞癌;增殖细胞核抗原;血管内皮生长因子;周期素D1;免疫组织化学     

Increased polycomb-group oncogene Bmi-1 expression correlates with poor prognosis in hepatocellular carcinoma

Purpose Recent studies have identified polycomb-group gene Bmi-1 as oncogene in the generation of mouse pre-cell lymphomas, and overexpression of Bmi-1 has been found in several human tumor with the disease progress and poor prognosis of the cancer patients. Methods In present study, we investigated Bmi-1 expression and its prognostic significance in hepatocellular carcinoma (HCC) by performing immunohistochemical analysis, using a total of 137 HCC clinical tissue samples. Results High Bmi-1 expression (Bmi-1 2+ or 3+) was shown in 29.9% cases. The positive immuno-staining of Bmi-1 was not only in well/moderately-differentiated tumor cells, but also in surrounding noncancerous or cirrhotic liver tissue. Bmi-1 expression level did not correlate with any clinicopathological parameters. However, survival analysis showed that the high-Bmi-1 group had a significantly shorter overall survival time than the low-Bmi-1 group (P = 0.047). Multivariate analysis after 24 months revealed that Bmi-1 expression was a significant and independent prognostic parameter (P = 0.002) for HCC patients. Conclusions Our study indicated that Bmi-1 could be a candidate biomarker for long-term survival in HCC. The first two authors contributed equally to this work.