存活素与血管内皮生长因子在甲状腺癌中的表达及其意义

目的探讨细胞凋亡抑制因子存活素在甲状腺癌中的表达效应及其与血管内皮生长因子(VEGF)表达的关系。方法采用免疫组织化学方法(SABC法)检测68例甲状腺癌、12例甲状腺腺瘤和10例正常甲状腺组织中存活素、caspase-3、VEGF蛋白表达,并结合甲状腺癌主要临床病理参数进行分析。结果存活素在正常甲状腺组织中未见表达,在甲状腺腺瘤中仅现微弱表达(16.7%),而在甲状腺癌中呈显著表达(57.4%);caspase-3在3种组织均有较高表达(70.0%、75.0%、69.1%),而VEGF在甲状腺癌和腺瘤中有不同程度表达(75.0%和41.7%);甲状腺癌中存活素与caspase-3表达无关,与VEGF表达显著正相关(r=0.302,P<0.05),且二者的表达在甲状腺癌组织学分型、临床分期及淋巴转移组间差异均有统计学意义。结论存活素特异性地表达于甲状腺癌组织中,可能通过阻抗细胞凋亡,并与VEGF协同促进血管生成,参与甲状腺癌的发生发展。     

Decreased immunoexpression of survivin could be a potential marker in human non‐alcoholic fatty liver disease progression?

Background/aim: Regulation of apoptosis in non‐alcoholic fatty liver disease (NAFLD) has been a theme of growing debate. Although no other study assessed the role of survivin in NAFLD, its expression has been reported in hepatic carcinogenesis because of other aetiological factors with relevant discrepancies. The aim of this study was to assess the pattern of survivin immunoexpression by tissue microarray along the whole spectrum of NAFLD, including non‐alcoholic steatohepatitis (NASH)‐related hepatocelular carcinoma (HCC). Methods: Liver biopsies from 56 patients with NAFLD were evaluated: 18 with steatosis, 21 non‐cirrhotic NASH, 10 NASH‐related cirrhosis, seven NASH‐related HCC, as compared with 71 HCC related to other causes and with 12 normal livers. Results: Survivin immunoexpression in NAFLD was restricted to cytoplasm and was found to be progressively lower in advanced stages, including cirrhosis and HCC: steatosis vs NASH‐related cirrhosis (P=0.0243); steatosis vs NASH‐related HCC (P=0.0010); NASH vs NASH‐related cirrhosis (P=0.0318); and NASH vs NASH‐related HCC (P=0.0007), thus suggesting a deregulation of apoptosis from NAFLD towards HCC. Interestingly, survivin immunoreactivity in NASH‐related HCC was also found to be significantly lower than in HCC related to other causes (P<0.05). Remarkably, nuclear staining for survivin was not detected in any case of NAFLD, contrasting to its presence in all other cases of HCC. Conclusions: Survivin immunoexpression in NASH‐related HCC is herein originally found substantially different than in HCC related to other causes, thus requiring further studies to elucidate the role of survivin in human NAFLD progression.     

靶向survivin的小干扰RNA对HCCLM6细胞恶性表型的影响

目的 研究survivin基因在肝癌细胞中的表达水平及survivin小干扰RNA对高侵袭潜能人肝癌细胞HCCLM6恶性表型的影响.方法 收集4种人肝癌细胞株,以RT-PCR和Western blot检测survivin蛋白质表达,进而筛选出高表达survivin基因的肝癌细胞.构建针对survivin mRNA的干扰质粒pshRNA-survivin及阴性对照质粒pGPU6/GFP/Neo-NC,并将其转染HCCLM6细胞.实时荧光定量PCR检测survivin mRNA表达量的变化情况;肿瘤细胞黏附实验检测细胞黏附能力的改变及Matrigel侵袭实验检测细胞侵袭力的改变. 结果 4种肝癌细胞随侵袭能力升高survivin mRNA表达水平依次升高(P＜0.05),HCCLM6细胞survivin表达水平最高;Western blot结果与RT-PCR结果一致.HCCLM6细胞转染pshRNA-survivin后,survivin mRNA表达量明显下降,抑制率达93.500%±3.117%,与转染阴性对照质粒组(8.215%±0.797%)相比,差异有统计学意义(t=70.852,P＜0.01).细胞黏附实验结果显示,survivin重组质粒转染组黏附率为11.403%±1.256%,阴性对照组为32.545%±1.367%(t=20.732,P＜0.01).Matrigel侵袭实验结果显示,与阴性对照组[(32.6±1.4)]个相比,转染pshRNA-survivin后,细胞侵袭力明显下降[(13.5±0.9)个,t=14.5,P＜0.01].结论 Survivin与肝癌侵袭转移等恶性生物学行为有关,并随侵袭转移潜能升高表达水平升高.pshRNA-survivin可以特异性抑制高侵袭肝癌细胞HCCLM6中survivin表达,并显著降低其侵袭、黏附能力,小干扰RNA技术有望成为抑制肝癌侵袭转移的新途径.     

肝细胞癌组织中骨桥蛋白的表达及其与血管生成的关系

目的探讨骨桥蛋白（OPN）在肝细胞癌（肝癌）组织中的表达与血管生成的关系。方法采用免疫组化法检测64例肝癌组织、10例肝硬化组织及10例正常组织中的OPN，分析其与肝癌临床病理特征和微血管密度（MVD）的关系。结果OPN在肝癌组织的表达明显高于肝硬化组织及正常组织（P〈0．05），OPN和肝癌的早期复发转移有关（P〈0．01），OPN的表达和MVD呈正相关（r=0．321，P〈0．01）。结论OPN在肝癌组织中里高表达与早期复发转移相关；OPN可能参与了肿瘤的血管生成。     

Immunohistochemical assessment of angiogenesis in hepatocellular carcinoma and surrounding cirrhotic liver tissues

AIM: To investigate whether vascular endothelial growth factor (VEGF) was over-expressed in hepatocellular carcinoma (HCC) or in surrounding cirrhotic liver tissues. METHODS: Immunohistochemistry was performed to investigate the expression of VEGF proteins in HCC tissues from 105 consecutive patients undergoing curative resection for HCC. The immunostaining results and related clinicopathologic materials were analyzed with statistical methods. Kaplan-Meier method was used to calculate survival curves, and Log-rank test was performed to compare differences in survival rates of the patients with positive HCC staining and negative VEGF. RESULTS: VEGF-positive expression was found in 72 of 105 HCC patients (68.6%). Capsular infiltration (P= 0.005), vascular invasion (P= 0.035) and intrahepatic metastasis (P=0.008) were observed more frequently in patients with VEGF-positive expression than in those with VEGF-negative.expression. Kaplan-Meier curves showed that VEGF-positive expression was associated with a shorter overall survival (P= 0.014). VEGF-positive expression was found in 47 of tissues 68 HCC (69.1%), and VEGF-positive expression was found in 54 of 68 surrounding cirrhotic liver tissues (79.4%). VEGF-positive expression was significantly higher in surrounding cirrhotic liver tissues than in HCC (P=0.017). CONCLUSION: VEGF may play an important role in the angiogenesis and prognosis of HCC, as well as in the angiogenesis of liver cirrhosis.     

生存素、p27和PTEN在肝细胞癌中的表达及临床病理意义

目的 探讨生存素蛋白、生存素mRNA、p27蛋白、p27 mRNA和第10号染色体同源丢失性磷酸酶张力蛋白基因(PTEN)蛋白在肝细胞癌(HCC)中的表达及其临床病理意义.方法 自制组织芯片,采用免疫组织化学和原位杂交法检测生存素蛋白、p27蛋白、PTEN蛋白和生存素mRNA、p27 mRNA在141份肝细胞癌、128份癌旁肝组织、97份远癌肝组织及17份正常肝组织中的表达,探讨各指标的关系并建立预测肝癌发生的模型.结果 肝癌组中生存素蛋白(Ridit值的95%CI为0.689±0.048,P《0.01)、生存素mRNA(Ridit值的95% CI为0.690±0.049,P《0.01)和p27蛋白(Ridit值的95% CI为0.556±0.053,P《0.05)表达明显增高,PTEN(Ridit值的95% CI为0.282±0.048)表达明显下降(P《0.01);肝癌中生存素的表达与p27、PTEN表达均显著相关;生存素mRNA、p27蛋白和PTEN蛋白对判断肝癌发生与否有重要意义.结论 生存素mRNA、p27蛋白的表达上调和PTEN蛋白的表达下调可作为判断肝癌发生的有价值指标.     

细胞间粘附分子-1在肝癌组织的表达及意义

目的：研究细胞间粘附分子－１（ＩＣＡＭ－１）在原发性肝癌的临床意义。方法：以免疫线化方法结合全自动图像分析检测ＩＣＡＭ－１在３０例原发性肝癌组织及６例正常肝组织的表达,分析其与肿瘤分化程度、大小及转移之间的关系。结果：３０例原发性肝癌组织２５例ＩＣＡＭ－１表达阳性（阳性率８３．３％）,肝癌组织中ＩＣＡＭ－１含量明显高于正常肝组织（Ｐ〈０．０１）,转移组肝癌中ＩＣＡＭ－１含量明显地无转移组（Ｐ〈０．     

Clinicopathological significance of expression of paxillin, syndecan-1 and EMMPRIN in hepatocellular carcinoma

AIM: To evaluate the relationship of expression of paxillin, syndecan-1 and EMMPRIN proteins with clinicopathological features in hepatocellular carcinoma (HCC). METHODS: Fifty-one patients who underwent HCC resection were recruited in the study. Paxillin, syndecan-1 and EMMPRIN proteins in HCC tissues were detected with immunohistochemical staining. RESULTS: Of 51 cases of HCC, 23 (45%) exhibited paxillin protein positive expression. Of 42 cases of adjacent non-tumor liver tissues, 24 (57%) exhibited positive expression. Positive paxillin protein expression was associated with low differentiation (r= 0.406, P= 0.004), with the presence of portal vein thrombosis (r = 0.325, P = 0.021), with extra-hepatic metastasis (r=0.346, P=0.014). Of 51 cases of HCC, 28 (55%) exhibited syndecan-1 protein positive expression. Of 42 cases of adjacent non-tumor liver tissues, 23 (55%) exhibited positive expression. Positive snydecan-1 protein expression was associated with well differentiation (r=0.491, P=0.001), with no extra-hepatic metastasis (r=0.346, P=0.014). Of 51 cases of HCC, 28 (55%) exhibited EMMPRIN protein positive expression. Of 42 cases of adjacent non-tumor liver tissues, 21 (50%) exhibited positive expression. Expression of EMMPRIN protein was not associated with serum AFP level, HBsAg status, presence of microsatellite nodule, tumor size, presence of cirrhosis and necrosis, differentiation, presence of portal vein thrombosis, extra-hepatic metastasis, disease-free survival and overall survival (P>0.05). Expression of paxillin protein was correlated conversely with the expression of syndecan-1 protein in HCC (r = -0.366, P = 0.010). CONCLUSION: Expression of paxillin and syndecan-1 proteins in HCC may affect its invasive and metastatic ability of the tumor. There may be a converse correlation between the expression of paxillin and syndecan-1 protein in HCC. Expression of EMMPRIN protein may be detected in HCC, but it may play little role in the invasion and metastasis of HCC.     

Plk1与Survivin蛋白在肝细胞癌中的表达及意义

目的探讨Plk1和凋亡抑制蛋白Survivin在HCC中表达情况,为临床诊断和治疗提供理论依据。方法采用免疫组化法检测100例肝细胞癌标本中Plk1、Survivin的表达。结果⑴肝细胞癌中,Survivin蛋白的表达明显高于癌旁正常组织（P〈0.05）,Survivin在肝细胞癌组织中的表达与患者性别、年龄、肿瘤分化程度、AFP表达及是否侵犯肝被膜均无关联（P〉0.05）。与HBsAg表达有关联（P〈0.05）。⑵肝细胞癌组织中,Plk1阳性表达明显高于癌旁正常组织（P〈0.05）;Plk1在肝细胞癌组织中的表达与患者性别、年龄、AFP表达、HBsAg表达及是否侵犯肝被膜均无关（P〉0.05）,与肿瘤分化程度相关（P〈0.05）。⑶肝细胞癌组织中,Plk1与Survivin表达呈正相关（P〈0.05）。结论 Plk1与Survivin的联合检测为肝细胞癌的诊断和治疗提供新的依据。     

Survivin expression in early hepatocellular carcinoma and post-treatment with anti-cancer drug under hypoxic culture condition

AIM:To investigate the expression of survivin during the early stages of hepatocellular carcinoma(HCC). METHODS:Immunohistochemical expression of survivin in liver tumor and non-tumor tissue specimens taken from 17 patients was compared.In addition,to determine the survivin expression in response to anti- cancer drugs in early stage HCC,the survivin expression was determined after the treatment of the HCC cells with anti-cancer drugs under hypoxic culture conditions. RESULTS:Survivin proteins were expressed in 64.7% of cells in early HCC specimens.A correlation between the survivin expression rate in the peritumoral hepatocytes and the rate of expression in the HCC specimens(low-rate group vs high-rate group)was observed.The survivin protein concentration in HCC cells was increased by the combination of hypoxia and anti-cancer drugs. CONCLUSION:This study suggests that survivin could be used as a therapeutic target in early HCC.     

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.     

Clinical value of apoptosis and angiogenesis factors in estimating the prognosis of hepatocellular carcinoma

Purpose Whereas some studies have indicated that the prognosis of hepatocellular carcinoma (HCC) was correlated to some apoptosis and angiogenesis factors: p53, survivin, matrix metalloproteinases (MMPs, including MMP-2 and MMP-9) and vascular endothelial growth factor (VEGF), other studies have failed to confirm this. The aim of the present study is to investigate the expression of p53, survivin, MMPs and VEGF in HCC and the relationship between these factors and the prognosis of HCC patients.Methods The expression of p53, survivin, MMP-2, MMP-9 and VEGF was measured by immunohistochemical assays in the liver resection specimens of 90 patients with HCC.Results The positive rate of p53, survivin, MMP-2, MMP-9 and VEGF was 33.3, 51.1, 60.0, 37.8 and 76.7%, respectively. The expression of MMP-2, MMP-9 and VEGF was correlated to the recurrence of HCC patients, respectively (P < 0.01). No correlation was found between the expression of apoptosis factors (p53 and survivin) and the recurrence of HCC patients, respectively (P > 0.05). The positive correlations were found between MMP-2 and VEGF (r = 0.32, P < 0.01), MMP-9 and VEGF (r = 0.24, P < 0.05). Significant differences of disease-free survival rates occurred among subgroups according to the expression of MMP-2, MMP-9 and VEGF (P < 0.01). Multivariate analysis revealed that macroscopically disseminated nodules, tumor micrometastasis, high serum alpha-fetoprotein level, positive expression of MMP-9 and VEGF were independent recurrence risk factors.Conclusions Our investigation revealed that p53 and survivin could not estimate the prognosis of HCC patients. Angiogenesis factors (MMPs and VEGF) positively correlated to the prognosis of HCC patients. The expression of MMPs and VEGF in HCC tissues could be regarded as a valuable indicator in estimating the prognosis of HCC patients.     

Expression of survivin in human gastric carcinoma and gastric carcinoma model of rats

AIM: To study the expression of survivin, an inhibitor of apoptosis protein, in human gastric carcinomas and gastric carcinoma models of rats.METHODS: With the method of immunohistochemical staining, we studied the expression of survivin in 20 cases of chronic gastritis and 56 cases of gastric carcinomas. We used N-methyI-N‘‘-nitro-N-nitrosoguanidine (MNNG) and high dose sodium-chloride diet to induce rat gastric carcinomas. Survivin expression was studied in glandular stomachs of normal rats, adenocarcinomas and tissues adjacent to the tumor, as well as in rats during the induction period.RESULTS: Survivin was expressed in 27 of 56 (48.2 %) cases of human gastric carcinoma tissues and 1 of 20 (5 %) cases of chronic gastritis. It was found that the expression of survivin had no relation with the elements of age, tumor depth, tumor size, and disease stage, but was significantly related to histological type. The positive rate of survivin expression in cases of intestinal type was significantly higherthan that in cases of diffuse type (P&lt;0.05). In animal experiments, survivin expression in glandular stomachs ofnormal rats, of rats in middle induction period, in adenocarcinomas and tissues adjacent to tumor were 0,40.0 %, 78.3 % and 38.9 %, respectively. Compared with the survivin expression in normal rats, the differences were significant.CONCLUSION: These data imply that survivin plays an important role in the onset of gastric carcinoma and that high survivin expression is an early event of gastric carcinoma.     

肝癌组织中survivin、VEGF的表达及意义

目的探讨survivin与血管内皮生长因子（VEGF）在肝细胞肝癌（HCC）组织中的表达及其临床意义。方法采用免疫组化法检测50例HCC组织和20例正常肝组织中survivin、VEGF蛋白的表达,并分析其与HCC临床病理参数的关系。结果50例HCC组织中survivin、VEGF蛋白的阳性表达率分别为64.0%、68.0%,在正常肝组织中的表达率分别为0、10%,两者相比,P均〈0.05;survivin、VEGF蛋白表达与HCC临床分期及淋巴结转移相关（P〈0.05）;HCC中survivin、VEGF蛋白表达呈正相关（r=0.886,P〈0.05）。结论survivin和VEGF蛋白在HCC中表达率较高,为肝癌的分子靶向治疗提供了新的靶点;survivin和VEGF在HCC中的表达关系密切,对肝癌的发展可能有协同作用;测定HCC中survivin、VEGF蛋白的表达,对判断患者预后及指导治疗有较大帮助。     

survivin在胃肠道间质瘤中的表达及其与COX-2表达的相关性

目的 探讨凋亡抑制蛋白survivin在胃肠道间质瘤(GISTs)组织中的表达情况及其与COX-2表达的相关性.方法 采用免疫组化S-P法检测57例胃肠道间质瘤组织中survivn蛋白和COX-2蛋白的表达情况.结果 57例GISTs中,survivin 38例阳性,阳性率为66.7%;COX-2为47例阳性,阳性率82.5%.survivin的蛋白阳性表达率与GISTs的恶性潜能相关,差异有统计学意义(P<0.01);而COX-2蛋白的阳性表达率与GISTs的恶性潜能无关(P>0.05);survivin表达与COX-2表达呈正相关(P<0.01).结论 survivin在胃肠道间质瘤组织中表达升高,并与GISTs的恶性潜能相关,可成为预测GISTs恶性潜能的指标.survivin与GISTs组织中COX-2的阳性密切相关,两者可能存在共同的激活机制,参与抑制GISTs的细胞凋亡.