肝细胞癌及癌旁组织中c—erbB—2—蛋白的表达与乙型肝炎病毒感 …

目的及方法 应用免疫组织化学技术检测了７６例肝细胞癌（ＨＣＣ）及其癌旁组织ｃ－ｅｒｂＢ－２蛋白和增殖细胞核抗原（ＰＣＮＡ）的表达，结果及结论 ｅ－ｅｒｂＢ－２蛋白在ＨＣＣ和癌旁组织内表达的阳性率分别为５７．９％（４４／７６）和７１．９％（４１／５７），阳性表达与ＨＣＣ组织学分级及细胞增殖状态无相关性，而与ＨＢＶ感染密切相关，ＰＣＮＡ在ＨＣＣ及癌旁组织阳性率分别为７７．６％（５９／７６）和３５．１％     

c－erbB－2及p53蛋白在原发性肝癌肝组织的表达

目的和方法应用免疫组化ＬＳＡＢ（Ｌａｂｌｌｅｄｓｔｒａｐｔａｖｉｄｉｎ－ｂｉｏｔｍ）法对４５例人原发性肝癌（ＰＨＣ）及其癌周肝组织以及５例正常肝组织进行了ｃ－ｅｒｂＢ－２和突变型ｐ５３癌基因蛋白的表达研究。结果ｃ－ｅｒｂＢ－２癌基因蛋白和ｐ５３蛋白在ＰＨＣ及其癌周肝硬变和非肝硬变组织中的阳性表达率分别是７１．１％，２１．４％（６／２８），１１．７％（２／１７）和４６．７％，１４．３％（４／２８），０％（０／１７），在５例非肝脏疾病的正常肝组织中均未见表达。ｃ－ｅｒｂＢ－２和ｐ５３蛋白二者表达强度与ＰＨＣ病理组织学分级具有一定程度相关关系，但与ＰＨＣ的不同组织学类型无显著差别。ｃ－ｅｒｂＢ－２与ｐ５３蛋白在原发性肝癌中的表达二者具有显著的相关性（Ｐ＜０．００５）。结论突变型ｐ５３和癌基因ｃ－ｅｒｂＢ－２在ＰＨＣ中具有独立和协同作用，对肝癌的发展具有重要影响。     

增殖细胞核抗原和C-erbB-2癌基因蛋白在乙肝病毒感染慢性肝病中表达的意义

对１４６例慢性乙型肝炎，肝硬化和肝细胞癌标本作了免疫染色，观察其ｃ－ｅｒｂＢ－２癌基因蛋白及增殖细胞核抗原的表达情况，探讨他们在ＨＢＶ诱发ＨＣＣ过程中的作用意义。结果：（１）在ＨＢＶ感染的慢性肝炎阶段已出现ｃ－ｅｒｂＢ－２Ｐ＾１８５和ＰＣＮＡ的异常表达；总阳性率分别为８７％和８．１％；（２）由慢性乙肝，肝硬化向肝细胞癌发展过程中，出现了ＰＣＮＡ表达逐渐升高，ｃ－ｅｒｂＢ－２Ｐ＾１８５逐渐下降的趋势     

老年大肠癌中PCNA和C-erbB-2的表达及意义

目的：检测ＰＣＮＡ和Ｃ－ｅｒｂＢ－２在老年大肠癌中的表达及其意义。对３３例老年大肠癌的石蜡切片标本进行免疫组化染色。结果ＰＣＮＡ和Ｃ－ｅｒｂＢ－２在３３例大肠癌中的阳性率分别为６６．７％和６０．６％;ＰＣＮＡ和Ｃ－ｅｒｂＢ－２的表达与癌组织的分化程度及临床分期有明显相关性（Ｐ〈０．０１）,淋巴结转移组中二者的表达明显高于无转移组（Ｐ〈０．０１）。结论ＰＣＮＡ和Ｃ－ｅｒｂＢ－２过高表达者预后不良,Ｐ     

差别PCR技术检测胃癌C一erbB-2癌基因扩增的临床意义

目的探讨胃癌Ｃ－ｅｒｂＢ－２癌基因扩增的临床意义。方法应用差别ＰＣＲ技术检测Ｃ－ｅｒｂＢ－２在８３例胃癌及１０１个转移淋巴结扩增情况。结果２８．９％（２４／８３）的胃癌存在该基因扩增，进展期胃癌及伴淋巴结转移者扩增阳性率增高（Ｐ＜０．０５及Ｐ＜０．０１）；转移淋巴结扩增阳性率明显高于胄癌原发灶（Ｐ＜０．０５）。早期胃癌及高中分化胃癌伴Ｃ－ｅｒｂＢ－２扩增者５年生存率低于不扩增者（Ｐ＜０．０５）。用Ｓｏｕｔｈｅｒｎｂｌｏｔ分子杂交及免疫组化ＡＢＣ法检测原胃癌样本基因扩增及过度表达，扩增及过度表达率分别为１５．７％及１８．１％，均明显低于上述扩增阳性率。结论差别ＰＣＲ技术检测Ｃ－ｅｒｂＢ－２扩增是一种快速方便、可靠、独立的方法，对胃癌预后及转移潜力是一种良好的判断指标。     

胃癌c—erbB—2及增殖细胞核抗原表达与预后的关系

目的：探讨胃癌ｃｅｒｂＢ２及增殖细胞核抗原（ＰＣＮＡ）表达与预后的关系。方法：应用免疫组化ＡＢＣ法对１０３例胃癌胃镜活检组织进行检测。结果：分别有１９例与５３例出现ｃｅｒｂＢ２阳性表达及高ＰＣＮＡ阳性标记（＞３５６１％），两者表达与胃癌大体类型、进展程度、组织类型、分化程度及有无伴淋巴结及肝转移情况密切相关。高ＰＣＮＡ阳性标记者ｃｅｒｂＢ２表达阳性率高，５年生存率低；高中分化胃癌中，ｃｅｒｂＢ２表达阳性者５年生存率显著低于阴性者。结论：提示ｃｅｒｂＢ２与ＰＣＮＡ表达检测可作为胃癌预后估计的独立指标     

原位聚合酶链技术检测肝细胞癌组织内丙型肝炎病毒核酸及其意义

应用一步反转录。聚合酶链原位扩增法（ＯＲＴ－ＰＣＲＩＳ）检测肝细胞癌（ＨＣＣ）及癌周组织内丙型肝炎病毒核糖核酸（ＨＣＶＲＮＡ），同时以一步反转录－ＰＣＲ（ＯＲＴ－ＰＣＲ）法检测血清及肝组织匀浆内ＨＣＶＲＮＡ。发现癌和癌周组织经原位扩增后ＨＣＶＲＮＡ检出率为７７．８％（２／２７）及８１．５％（２２／２７），而血清及组织匀浆内仅为２９．６％及３７．Ｏ％，两者与癌及癌周组织相比差异均有显著性（Ｐ＜０．０１）。ＯＲＴ－ＰＣＲＩＳ示在癌组织中ＨＣＶＲＮＡ以核型为主（１２／２１），阳性细胞点样分布，细胞内信号强度多为＋（５７．１％）；而癌周组织以核浆型为主（１４／２２），阳性细胞弥漫分布，信号强度多为＋＋＋（５０．０％）。无论在癌还是在癌周组织均未发现ＨＣＶｃＤＮＡ存在。结果提示ＯＲＴ－ＰＣＲＩＳ优于ＲＴ－ＰＣＲ，ＨＣＶ对本地区ＨＣＣ发生、发展起着极为重要的作用。癌周ＨＣＶ复制较癌组织内活跃，癌和癌周细胞核阳性表明ＨＣＶＲＮＡ在核内整合或复制，这种整合或复制对肝细胞基因组ＤＮＡ产生的影响可能与乙型肝炎病毒（ＨＢＶ）相类似。     

MAGE-1基因在肝细胞性肝癌中的表达

目的探讨ＭＡＧＥ－１基因编码的肿瘤相关抗原作为肝细胞性肝癌（ＨＣＣ）主动免疫治疗攻击靶点的可能性。方法用ＲＴ－ＰＣＲ的方法检测了３９例ＨＣＣ及癌旁组织、３种人肝癌细胞株和５例非ＨＣＣ病例肝组织中ＭＡＧＥ－１基因的表达。结果ＨＣＣ癌组织中ＭＡＧＥ－１基因表达率明显高于癌分组织中的表达率,５８．９％（２／３９）对１０．３％（４／３９）,Ｐ＜００１。３种细胞株均为阳性表达。结论ＭＡＧＥ－１基因可作为主动免疫治疗的攻击靶点,以进一步研究设计新的ＨＣＣ免疫治疗策略。     

bcl—2蛋白在肝肿瘤组织的表达及意义

目的 探讨抗凋亡基因ｂｃｌ－２蛋白在常见肝肿瘤发生中的作用。方法 应用ｂｃｌ－２癌蛋白单克隆抗体对６４例四种类型的肝恶性肿瘤进行ＬＳＡＢ免疫组织化学标记。结果 １６．２％（６／３７）的肝细胞癌，６９．２％（９／１３）的胆管细胞癌，２０．０％（１／５）的肝母细胞瘤和２２．２％（２／９）肝转移癌呈ｂｃｌ－２癌蛋白阳性表达，其中胆管细胞癌的阳性率显著高于其癌旁组织（Ｐ值〈０．０５）和肝细胞癌（Ｐ值〈０．     

肝癌及癌旁组织HBV DNA定点PCR研究

目的采用定点ＰＣＲ技术，研究ＨＢＶ的Ｓ和Ｃ基因在肝癌与癌旁组织的存在状态，探讨癌旁ＨＢｓＡｇ表达与Ｓ基因的关系。方法福尔马林固定，石蜡包埋肝癌及癌旁组织２８例，ＨＢｓＡｇ免疫组化后的切片分别切割癌组织、癌旁ＨＢｓＡｇ强阳性区（Ｐ２）及癌旁ＨＢｓＡｇ阴性区（Ｐ１）各１个低倍视野，作Ｓ及Ｃ基因ＰＣＲ。结果癌组织Ｓ基因较Ｃ基因检出率高（１８／２８ｖｓ１０／２８，Ｐ＜０．０５），癌旁二者无差异；癌旁中Ｐ１与Ｐ２的Ｓ及Ｃ基因检出率无显著性差异；高分化肝癌Ｓ和／或Ｃ基因检出率较中分化者低（２／５ｖｓ２０／２８，Ｐ＜０．０５）。结论癌组织中整合的ＨＢＶＤＮＡ为残缺不全的，ＨＢｓＡｇ强阳性与阴性区的Ｓ基因检出无显著性差异。     

Immunohistochemical study on p53, H-rasp21, c-erbB-2 protein and PCNAexpression in tumor tissues of Hah and minority ethnic patients with primaryhepatic carcinoma in Xinjiang

AIM To find out if there is any difference in human primary liver carcinogenesis between Han and minorityethnic patients in Xinjiang.METHODS Expression of p53, c-erbB-2, H-rasp21 protein and proliferating cell nuclear antigen (PCNA)in tumor tissues of 50 patients (Hah 38, minorities 12) with primary hepatic carcinoma (HCC) was detectedby immunohistochemistry (LSAB).RESULTS The positive frequency of p53, c-erbB-2, H-rasp21 and PCNA expression was 46.0% (23/50,70.0% (35/50), 68.0% (34/50) and 82.0% (41/50) in tumor tissues; 4.0% (2/50), 22.0% (11/50),64.0% (32/50) and 52.0% (26/50) in peritumor respectively with a significant difference, except for H-rasp21 (P<0.05) between tumor and non-tumor tissues. Combined the three oncogenes alteration, 26%(13/50) tumor tissues had positive immunoreactivity, but peritumor and normal liver were negative. Thepositive p53, c-erbB-2, H-rasp21 protein expression was 39.5 % ( 15 / 38), 60.5 % (23 / 38) and 39.5 % ( 15 /38) in tumors of Han patients; 66.7% (8/12), 100% (12/12) and 75.0% (9/12) in minority patientsrespectively. A statistical difference between Han and minority cancer samples was observed (P< 0.05).CONCLUSION Overexpression of p53, c-erbB-2 and H-rasp21 in human primary liver carcinoma is animportant biomarker of genetic alteration. The different frequency of these oncogenetic changes may reflectsome environmental factors or/and ethnic hereditary affecting the liver carcinogenesis. The special life styleof Han, Uygur, Kazak and Mongolia nationalities in Xinjiang may also involve the etiopathogenesis of thisdisease.     

Immunohistochemical detection of proliferating cell nuclear antigen in hepatocellular carcinoma

ＩｍｍｕｎｏｈｉｓｔｏｃｈｅｍｉｃａｌｄｅｔｅｃｔｉｏｎｏｆｐｒｏｌｉｆｅｒａｔｉｎｇｃｅｌｎｕｃｌｅａｒａｎｔｉｇｅｎｉｎｈｅｐａｔｏｃｅｌｕｌａｒｃａｒｃｉｎｏｍａＷＡＮＧＤｏｎｇ１，ＳＨＩＪｉｎｇＱｕａｎ２ａｎｄＬＩＵＦｅｎｇＸｕａｎ３Ｓｕ...     

Cell proliferation, apoptosis and the related regulators p27, p53 expression in hepatocellular carcinoma

AIM: To investigate the expression of cell apoptosis, proliferation and the related regulators p27,p53 in hepatocellular carcinoma (HCC). METHODS: The expression of p27, p53, proliferating cell nuclear antigen (PCNA) and apoptosis in 47 HCC specimens and 42 surrounding non-cancerous tissues were detected by the immunohistochemistry and terminal deoxy-nudeotidyl transferase-mediated nick end labeling (TUNEL) technique. Meanwhile, the clinical significance of them was analyzed combining with the clinicopathological factors and follow-up data. RESULTS: (1) The average proliferating index and apoptotic index in HCC were significantly higher than that in adjacent liver tissues. The proliferating index was associated with extrahepatic metastasis. The apoptotic index was significantly lower in TNM stage Ⅰ-Ⅱ than in stage Ⅲ-Ⅳ. The proliferating index of groups with p53-/p27+ was significantly lower than that in group with p53+/p27- (P= 0.030); (2) The level of p27 in the cytoplasmic fraction was higher in non-tumoral liver tissues and was associated with clinical stage; (3) Survival analysis showed advanced stage (P=0.031) and with extrahepatic metastasis (P = 0.045) was significantly associated with shorter survival. In addition, the prognosis of patients with p53-/p27+ was longer than that of patients with p53+/p27- (P= 0.0356). CONCLUSION: The p53 mutation and decreased p27 expression might be involved in the imbalance of proliferation and apoptosis in HCC. Cytoplasmic displacement might lead to the inactivation of p27 protein in HCC cells and acts early during carcinogenesis of HCC. The combined examination of p27, and p53 expression allows reliable estimation of prognosis for patients with primary hepatic carcinoma.     

Prognostic value of proliferating cell nuclear antigen and c-erbB-2 compared with conventional histopathological factors in breast cancer

Expression of proliferating cell nuclear antigen (PCNA) and c-erbB-2 oncoprotein has been assessed in 471 women with breast cancer to evaluate their prognostic value as compared to conventional histopathological factors. In univariate analysis, high PCNA expression (20%) predicted a significantly worse survival in lymph-node-negative tumors (univariateP=0.031). However, the effect disappeared in multivariate analysis and the histological grade remained the only independent factor for this group. Despite its close correlation to histological grade (P<0.001), PCNA expression discriminated subsets with different survival within the heterogeneous group of moderately differentiated tumors (univariateP=0.073, multivariateP=0.075). PCNA expression was not found to be a significant prognostic factor in lymph-node-positive tumors, thus it was of limited value for breast cancer patients as a whole. c-erbB-2 protein overexpression was associated with a worse survival (univariateP=0.019, multivariateP=0.057) for the entire group of patients. The effect was mainly attributed to the significance of c-erbB-2 as an independent factor in lymph-node-positive (up to three nodes, multivariateP=0.04; four or more nodes: multivariateP=0.017) and large tumors (>2 cm: multivariateP=0.002). c-erbB-2 was without significance in lymph-node-negative patients. Though both factors might amplify the prognostic information for distinct patient subsets they do not achieve the strong prognostic value of conventional histopathological features in breast cancer.     

Expression of p57(kip2) and its relationship with clinicopathology, PCNA and p53 in primary hepatocellular carcinoma

AIM: To investigate the expression of p57kip2 and its relationship with clinicopathology, PCNA and p53 in primary hepatocellular carcinoma (HCC). METHODS: Expression of p57kip2, PCNA and p53 in tumor tissues from 32 patients with HCC and 10 liver tissues of normal persons was detected with Elivision immunohistochemical technique. RESULTS: The p57kip2 protein positive-expression rate in HCC was 56.25%, lower than that in normal tissues (100%, P<0.05). The reduced expression of p57kip2 protein correlated significantly with moderate or low differentiation of tumor cells (P = 0.007 <0.05), high clinical stage (P= 0.041 <0.05) and poor prognosis (P= 0.036 <0.05), but did not correlate significantly with metastasis, tumor size, level of AFP and age (P>0.05). The PCNA positive-expression rate was 56.25%, which was correlated significantly with the expression of p57kip2 (P= 0.025<0.05). The p53 positive-expression rate was 46.88%, which was not correlated significantly with the expression of p57kip2 (P>0.05). CONCLUSION: There is a marked loss or absence of p57kip2 expression and high expression of PCNA in HCC, which are involved in carcinogenesis and development of HCC. The p57kip2 and p53 may induce apoptosis via different mechanisms.     

Expression of transforming growth factor-alpha and hepatitis B surface antigen in human hepatocellular carcinoma tissues and its significance

AIM:To evaluate the expression of transforming growthfactor-alpha (TGF-α) and hepatitis B surface antigen (HBsAg) in human hepatocellular carcinoma (HCC) tissues and its significance.METHODS:Seventy specimens of HCC tissues were detected by immunohistochemical method. Five specimens of normal human liver tissues were used as control.RESULTS: The TGF-o~ positive expression rates in HCC and its surrounding tissues were 74.3%(52/70) and 88.1%(52/59), respectively. TGF-α positive granules were mainly in the cytoplasm and fewer existed on the karyotheca. The TGF-α positive expressing rate in well differentiated HCC was significantly higher than that in moderately and poorly differentiated HCC (P&lt;0.05).The TGF-α positive expression also was observed in intrahepatic bile ducts (part of those were hyperplastic ducts).The HBsAg positive expression rates in HCC and its surrounding tissues were 21.4%(15/70) and 79.7%(47/59), respectively.HBsAg positive granules were in the cytoplasm, inclusion and on the karyotheca.There was a prominent positive correlation between TGF-α and HBsAg expression in HCC surrounding tissues (P&lt;0.05,γ=0.34). TGF-α was usually existed with HBsAg in regenerated and/or dysplastic liver cells.In the five normal liver tissues, TGF-α and HBsAg were not detectable in hepatocytes and bile ducts.CONCLUSION:Hepatitis B virus infection is dosely related with hepatocarcinogenesis.The overexpression of TGF-α in the liver seems to be associated with the regeneration of hepatocytes injured by HBsAg.The continued expression of TGF-α might lead to dysplasia of liver cells and development of HCC. Furthermore, TGF-α might play a role in morphogenesis and regeneration of intrahepatic bile ducts.     

Expression of transforming growth factor-α and hepatitis B surface antigen in human hepatocellular carcinoma tissues and its significance

AIM: To evaluate the expression of transforming growth factor-alpha (TGF-α) and hepatitis B surface antigen (HBsAg) in human hepatocellular carcinoma (HCC) tissues and its significance. METHODS: Seventy specimens of HCC tissues were detected by immunohistochemical method. Five specimens of normal human liver tissues were used as control. RESULTS: The TGF-α positive expression rates in HCC and its surrounding tissues were 74.3%(52/70) and 88. t%(52/59), respectively. TGF-α positive granules were mainly in the cytoplasm and fewer existed on the karyotheca. The TGF-α positive expressing rate in well differentiated HCC was significantly higher than that in moderately and poorly differentiated HCC (P＜0.05). The TGF-α positive expression also was observed in intrahepatic bile ducts (part of those were hyperplastic ducts). The HBsAg positive expression rates in HCC and its surrounding tissues were 21.4%(15/70) and 79.7%(47/59), respectively. HBsAg positive granules were in the cytoplasm, inclusion and on the karyotheca. There was a prominent positive correlation between TGF-α and HBsAg expression in HCC surrounding tissues (P＜0.05, γ=0.34). TGF-α was usually existed with HBsAg in regenerated and/or dysplastic liver cells. In the five normal liver tissues, TGF-α and HBsAg were not detectable in hepatocytes and bile ducts.CONCLUSION: Hepatitis B virus infection is closely related with hepatocarcinogenesis. The overexpression of TGF-α in the liver seems to be associated with the regeneration of hepatocytes injured by HBsAg. The continued expression of TGF-α might lead to dysplasia of liver cells and development of HCC. Furthermore, TGF-α might play a role in morphogenesis and regeneration of intrahepatic bile ducts.     

Relationship between hepatocyte proliferation and hepatitis delta virus replication in neoplastic and non-neoplastic liver tissues

SUMMARY. We studied the relationship between hepatocyte proliferation and hepatitis delta virus (HDV) replication at the single cell level. The proliferating cell nuclear antigen (PCNA) (by immunohistochemistry) and the HDV RNA (by in situ hybridization) were stained in neoplastic and non-neoplastic liver tissues of 19 patients with chronic HDV infection, including four cases of cirrhosis with superimposed hepatocellular carcinoma (HCC). As controls, we assessed the hepatocyte proliferation of liver tissues from 16 patients with chronic hepatitis B and on three normal livers. The hepatocyte PCNA labelling index of HDV-infected tissues was comparable with that seen in chronic hepatitis B-infected livers but was significantly higher than that observed in normal livers. Although cirrhotic tissues had lower hepatocyte proliferating fractions than non-cirrhotic tissues, the difference was not statistically significant. The hepatocyte proliferation rate did not correlate with the level of intrahepatic HDV replication or with the histological activity. In double-labelling experiments, PCNA and HDV RNA staining did not co-localize, with the exception of two of three cirrhotic tissues associated with HCC, where the association between the two markers was statistically significant. This co-localization was not observed, however, in the adjacent tumorous tissues. In patients with chronic HDV infection the hepatocyte proliferation was increased with respect to normal liver tissue, but was comparable with that observed in patients with chronic hepatitis B virus infection and did not correlate with the level of HDV replication or the histological activity. In the cirrhotic tissue of patients with HCC (but not in the tumour counterpart), HDV RNA may occasionally co-localize with the marker of hepatocyte proliferation. Whether this association between viral replication and cell division is related to liver carcinogenesis remains to be established.     

p21和增殖细胞核抗原在肝硬化组织的表达及其与肝细胞不典型增生的关系

目的探讨p21和增殖细胞核抗原（proliferatingcellnuclearantigen,PCNA）的相互关系及它们在肝硬化、肝细胞不典型增生（LCD）、肝细胞癌(HCC)形成中的作用。方法运用S-P法对60例单纯肝硬化、30例癌旁肝硬化及27例HCC的p21、PCNA及HBsAg表达情况进行实验研究。结果癌旁肝硬化、单纯肝硬化、癌组织p21阳性率分别为90%、68.33%、62.96%。癌旁肝硬化与单纯肝硬化及癌组织之间,差别有显著性意义(P＜0.05)。伴LCD改变的肝硬化,p21、PCNA及HBsAg阳性率分别为92.45%、73.59%、64.92%均高于不伴LCD改变的肝硬化组织（P＜0.05）。HBsAg阳性及PCNA表达阳性的肝硬化组织,其p21阳性表达率分别为87.32%、86.21%,均明显高于HBsAg阴性及PCNA阴性的肝硬化组织（P＜0.01）。结论(1)p21的过度表达可能与HCC的起始过程有关,在HCC形成发展的早期阶段发挥重要作用。(2)LCD是一群具有肿瘤性增殖潜能的癌前细胞群,尤其是伴有HBV感染,且有p21及PCNA共同过量表达者,有发生HCC的高度危险。(3)HBV感染及PCNA过量表达与p21表达密切相关。