THE HBx PROTEIN EXPRESSION IN LIVER CANCER

The expression of HBx protein in liver tissues from 48 cases of different liver diseases, Including 32 cases of hepatocellular carcinoma (HCC), 10 of chronic hepatitis (CH), 2 of angioma and 4 cases of normal liver was studied. These samples were tested for HBx protein, HBsAg by modified ABC method. Positive rates of HBx in cancer and adjacent Uver tissue were 75. 0% and 62. 5%, and positive rates of HBsAg were 37.5% and 78.1% respectively. The occurence of HBx in the absence of HBsAg was more frequently observed in tissues from HCC (46.9%) man CH (0%). The results showed mat expression of HBx was more active than that of HBsAg, and it Is suggested that HBx might be a useful marker for the diagnosis of liver cancer.     

EXPRESSION OF INSULIN-LIKE GROWTH FACTOR Ⅱ(IGF-Ⅱ)IN HUMAN HEPATOCELLULAR CARCINOMA AND LIVER CIRRHOSIS:ITS RELATIONSHIP WITH HEPATITIS B VIRUS X PROTEIN EXPRESSION

Sixty cases of hepatocellular carcinoma (HCC) and47 cases of liver cirrhosis (LC) were examined withimmunocytochemistry method using antibodies againstIGF-II and HBxAg on formalin-fixed, paraffin-embeddedtissue sections. 32 HCC and 37 LC were found to bepositive to HBxAg, in which the positive rates of IGF-IIwere 100% (32/32) and 94.6% (35/37) respectively. 28HCC and 10 LC were found to be HBxAg negative, IGF-IIwas positive in 23 HCC (83.1%) and 6 LC:(60%) Thepositive expression rates of IGF-II in HBxAg positivetissues were significanfly higher than those in HBxAgnegative tissues (P<0.05). There were three types ofdistribution of IGF-II expression in HCC and LC (1)perinucleus: (2) diffuse in cytoplasm; (3) inside nucleus.IGF-II was highly expressed in most of hyperplastic andneoplastic nodules hepatocytes and some of regenerationnodules. Small polygonal liver cells (SPLCs) were foundin the liver tissues surrounding the tumor and cirrhosisand they were positive to both IGF-II and HBxAg. Thepositive rates     

乙肝病毒相关的肝癌p16异常甲基化

Objective： To investigate the potential linkage between high rate of p16 methylation and hepatitis B virus （HBV） infection, methylation status of p16, HBV infection markers in serum and HBV-DNA replication level in cancerous and non-cancerous tissue of 32 cases of hepatocellular carcinomas （HCC） with HBV infection and 12 HCCs without HBV infection were examined. Methods： p16 methylation was detected with methylation-specific polymerase Chain reaction （PCR）, and HBV markers were examined with real-time PCR and immunologic method. Results： Methylation of p16 promoter was found in 31 （70.5%） of 44 cancerous tissues of HCC, 2 （16.7%） of 12 HCC without HBV infection, 29 （90.6%） of 32 HCCs with HBV infection marker, p16 methylation was detected in 5 （83.3%） of 6 HCCs positive for HBsAg and HBeAg, 17 （94.4%） of 18 HCCs positive for HBsAg and negative for HBeAg, 7/8 （87.5%） of HCCs positive for other HBV infection markers, such as HBsAB, HBcAb, HBeAb. p16 methylation products were also found in non-cancerous tissues of 4 cases of HCCs with HBV infection, not detected in non-cancerous tissues without HBV infection. HBV-DNA was detected in cancerous tissues of 29/32 （90%） HCCs with HBV infection. Surprisingly, Methylation product of p16 promoter was found in all cases （29/29） of HCCs with detectable HBV-DNA in neoplastic tissue. Conclusion： Persistent HBV infection may promote p16 hypermethylation, suggesting that HBV, via enhancing the aberrant methylation of p16, indirectly involved in development of HCC.     

C-erbB-2,p53,N-ras EXPRESSION IN HEPATOCELLULAR CARCINOMA

To assess the relationship between C-erbB-2, p53,N-ras status at a premalignant stage and in HCC, theauthors studied the imunohistological expression of thesegenes in HCC, liver cirrhosis and in the adjacent normalresected Iiver tissue, using monoclonal antibody tomutated p53 and activated C-erbB-2, N-ras. C-erbB-2was expressed in 97.1 % (35/36) of HCC and 100% (18/18)of helpatic cirrhosis, low level C-erbB-2 expression wasobserved in 2/14 (14.3% of normal liver specimens;The positive incidence of overexpression of mutant p53protein in HCC and hepatic cirrhosis were 55.6%(20/36)and 66.7%(10/18) respectively; 29 (76.5%) specimens ofHCC and 16 (88.9%) of hepatic cirrhosis were positive forN-ras protein. The overexpression of the three oncogeneproteins were significantly higher than that of normalliver tissues (P<0.01). These results indicated thatactivated C-erbB-2, N-ras and altered p53 genes may havea role in human HCC Pathogenesis through promiting thedevelopment of HCC from hepatic cirrhosis and thepro     

涎腺腺样囊性癌中丝裂原激活蛋白激酶p38的表达及意义

目的 探讨丝裂原激活蛋白激酶p38(p38MAPK)在涎腺腺样囊性癌(SACC)中的表达及临床意义.方法 采用组织微阵列平台,运用免疫组化和原位杂交技术,检测52例SACC和11例正常涎腺组织中p38MAPK蛋白和mRNA的表达,并分析其与SACC 临床病理特征的关系.结果 正常涎腺组织和SACC组织中,p38MAPK蛋白的阳性表达率分别为36.4%和96.2%,差异有统计学意义(P＜0.01).p38MAPK蛋白表达与SACC的淋巴结转移、神经侵犯有关(均P＜0.05).正常涎腺组织和SACC组织中,p38MAPK mRNA的阳性表达率分别为45.5%和94.2%,差异有统计学意义(P＜0.01).p38MAPK mRNA表达与SACC的淋巴结转移、神经侵犯有关(均P＜0.05).SACC组织中,p38MAPK蛋白与mRNA的表达呈正相关(r=0.409,P＜0.01).结论 在SACC组织中,p38MAPK表达上调,p38MAPK通路可能与SACC的发生、侵袭和转移有关.

Abstract：

Objective To investigate the expression of p38 mitogen-activated protein kinase (p38MAPK) in salivary adenoid cystic carcinoma (SACC) tissues and their clinicopathologic significance.Methods The protein and mRNA expressions of p38MAPK were examined by immunohistochemistry and in situ hybridization, respectively, in 52 cases of SACC and in 11 normal salivary gland tissues adjacent to the tumors on a tissue microarray platform.Results The positive rate of p38MAPK protein expression in the paracancerous normal salivary gland tissues and that in SACC were 36.4% and 96.2%, respectively,showing a significant statistical difference (P ＜ 0.01 ).The protein expression of p38MAPK was positively correlated with the lymph node metastasis and nerve involvement (P ＜ 0.05 ).The positive rates of p38MAPK mRNA in the paracancerous tissues and in the SACC tissues were 45.5% and 94.2%,respectively, with a significant statistical difference (P ＜0.01 ).The mRNA expression of p38MAPK was positively correlated with the lymph node metastasis and nerve involvement (P ＜ 0.05 ).In the SACC, there was a notable positive correlation between the p38MAPK protein and mRNA expressions (r = 0.409, P ＜0.01).Conclusions The expression of p38MAPK is up-regulated in salivary adenoid cystic carcinoma.p38MARK may be involved in the tumorigenesis, development and metastasis of this cancer.     

Expression of Inducible Nitric Oxide Synthase, p53 and Bcl-2 in Gastric Precancerous and Cancerous Lesions： Correlation with Clinical Features

OBJECTIVE To explore the expression of inducible nitric oxide synthase (iNOS), p53 and bcl -2 in gastric precancerous and cancerous lesions and to examine the expression of these proteins in relation to clinical features. METHODS The expressions of iNOS, p53 and bcl-2 proteins in gastric precancerous and cancerous lesions and their correlations with the clinical features were determined using immunohistochemical assays (Power VisionTM two -step method) on 84 gastric carcinomas and 54 gastric atypical hyperplastic tissues. Apoptotic cells were evaluated by terminal deoxynucleotidyl transferase- mediated dUTP-biotin nick-end labeling (TUNEL). RESULTS Expression of iNOS, p53 and bcl-2 was significantly higher in gastric carcinoma (GC) tissues than in gastric atypical hyperplastic tissues. Among the 84 carcinomas, the expression of p53 was observed in 50 (59.52%), bcl-2 in 43 (51.19%), and iNOS in 65 (77.58%). Overex-pression of iNOS and bcl-2 in gastric carcinoma was related to tumor size and iNOS was related to the presence of lymph node metastasis (P< 0.05). The expression of proteins did not correlate with age, sex, stage of disease, or differentiation. Expression of iNOS in gastric carcinoma tissues was positively correlated with bcl-2 expression (X2=8.926, P=0.003), and also with p53 expression (X2= 5.2430, P= 0.022). The mean apoptotic indexes (Al) were 1.29%±0.50 in low-grade atypical hyperplasia (LG), 0.96%±0.36 in high-grade atypical hyperplasia (HG) and 0.70%±0.43 in GC, with the difference being significant between LG, HG and GC (P< 0.05). There was a significant positive correlation between iNOS expression and the Al in GC (t=3.0815, P=0.0028). CONCLUSION iNOS was expressed in the majority of gastric carcinoma tissues and correlated with cellular apoptosis associated with p53 and bcl -2 expression. iNOS overexpression is closely associated with p53 and bcl-2 accumulation status. iNOS may play a synergistic role in the pathogenesis of GC.     

肝癌组织中Raf激酶抑制蛋白和磷酸化Raf 激酶抑制蛋白的表达及其意义

 目的探讨Raf激酶抑制蛋白（Raf kinase inhibitor protein,RKIP）和磷酸化Raf激酶抑制蛋白（Phosphorylated raf kinase inhibitor protein,P-RKIP）在肝细胞癌中的表达及其临床意义。方法应用组织芯片和免疫组织化学方法检测RKIP蛋白和磷酸化的RKIP蛋白在72例肝细胞癌组织、50例癌旁肝组织及16例正常肝组织中的表达,并分析RKIP的表达与肝细胞癌临床病理学特征的关系。采用Western blot方法检测36例肝细胞癌、36例癌旁组织及12例正常肝组织中RKIP蛋白和磷酸化的RKIP蛋白的表达。结果免疫组化结果显示肝细胞癌、癌旁及正常肝组织中RKIP的阳性率分别为22.2％、86％、93.8％,P-RKIP为29.2％、84％、93.8％;肝癌组织中RKIP和P-RKIP的表达显著低于癌旁及正常肝组织,差异有统计学意义（P<0.05）,而且肝癌组织和癌旁肝组织中两者蛋白的表达均呈显著正相关（P=0.000,P=0.005）;RKIP和P-RKIP的表达均与肝癌有无肝内或淋巴结转移及分化程度有关（P<0.05）。Western blot结果显示肝癌组织中RKIP和P-RKIP的表达显著低于癌旁及正常肝组织。结论RKIP和 P-RKIP表达的减少或缺失与肝癌的发生发展、侵袭转移密切相关。     

原发性肝细胞癌p21WAF1和p16表达及意义

[目的]研究p21WAF1、p16在原发性肝细胞癌(HCC)中的表达及意义.[方法]利用免疫组织化学技术检测74例HCC中p21WAFI、p16蛋白表达.[结果]p16蛋白在HCC中表达的阳性率为51.35%,p21WAF1蛋白的表达率为43.24%,而正常肝组织中为100%,两组均有显著性差异(P＜0.05).p16蛋白在有转移的HCC中阳性率为23.08%,明显低于无转移HCC的阳性率(71.43%,P＜0.05).[结论]p16及p21WAF1蛋白的缺失与HCC的发生发展有关.p16蛋白的缺失与HCC转移有关.     

RELATIONSHIP OF THE EXPRESSION OF MDR1 GENE PRODUCT IN HEPATO CELLULAR CARCINOMA TO INVASION AND METASTASIS

ＩＮＴＲＯＤＵＣＴＩＯＮＰｇｌｙｃｏｐｒｏｔｅｉｎ（Ｐｇｐ），ｔｈｅｐｒｏｄｕｃｔｏｆｔｈｅｍｕｌｔｉｄｒｕｇｒｅｓｉｓｔａｎｃｅ（ＭＤＲ１）ｇｅｎｅ，ｉｓａ１７０Ｋｄａｍｅｍｂｒａｎｅｐｒｏｔｅｉｎｒｅｓｐｏｎｓｉｂｌｅｆｏｒｐｕｍｐｉｎｇ...     

Increased protein expression of DNA methyltransferase (DNMT) 1 is significantly correlated with the malignant potential and poor prognosis of human hepatocellular carcinomas

Alteration of DNA methylation is one of the most consistent epigenetic changes in human cancers. DNA methyltransferase (DNMT) 1 is a major enzyme involved in establishing genomic methylation patterns. Most of the studies concerning DNMT1 expression in human cancers have been performed only at the mRNA level. To directly examine DNMT1 protein expression levels during human hepatocarcinogenesis, 16 histologically normal liver tissues, 51 noncancerous liver tissues exhibiting chronic hepatitis or cirrhosis, which are considered to be precancerous conditions, and 53 hepatocellular carcinomas (HCCs) were subjected to immunohistochemic examination. If more than 20% of the cells exhibited nuclear DNMT1 staining, the tissue sample was considered to be DNMT1-positive. DNMT1 immunoreactivity was observed in 23 (43%) of the HCCs, but in none (0%) of the histologically normal liver or noncancerous liver tissues exhibiting chronic hepatitis or cirrhosis. The incidence of increased DNMT1 protein expression in HCCs correlated significantly with poor tumor differentiation (p = 0.0006) and portal vein involvement (p = 0.0002). Moreover, the recurrence-free (p = 0.0001) and overall (p < 0.0001) survival rates of patients with HCCs exhibiting increased DNMT1 protein expression were significantly lower than those of patients with HCCs that did not exhibit increased expression. Increased DNMT1 protein expression may play a critical role in the malignant progression of HCCs and be a biologic predictor of both HCC recurrence and a poor prognosis in HCC patients.     

HIF-1α和VEGF 的表达与肝细胞癌侵袭转移的关系

 目的 探讨HIF-1α、VEGF的蛋白表达与肝细胞癌血管新生及侵袭转移特性之间的关系。方法 采用免疫组化SP法检测肝细胞癌、癌旁肝组织和正常肝组织中HIF-1α、VEGF的表达，并计数MVD值。结果 HIF-1α在肝细胞癌中的阳性表达率为50．0％，显著高于癌旁（16．1％）和正常肝组织的（8．3％）（P=0．001）；VEGF在肝细胞癌中的阳性表达率为81）％，显著高于癌旁（59．4％）和正常肝组织（41．7％）（P=0．044）；肝细胞癌组织中MVD值显著高于癌旁和正常肝组织（P=0．001）。HIF-1α蛋白表达与有无门静脉或胆管癌栓及肿瘤分化程度有关（P〈0．05）；VEGF蛋白的表达与有无肝内或淋巴结转移及有无门静脉或胆管癌栓有关（P〈0．05）。HF-1α与VEGF表达有关（P=0．005）；HF-1α与VEGF共同表达阳性组的MVD值显著高于共同表达阴性组MVD值（P=0．001）。结论 HF-1α可能通过调节VEGF的表达促进肝细胞癌血管新生，从而促使肝细胞癌侵袭转移。     

pERK在胃癌中的表达及临床意义

[目的]研究磷酸化细胞外信号调节激酶（pERK）在胃腺癌、慢性萎缩性胃炎及浅表性胃炎组织中的表达及意义。[方法]RT-PCR法检测胃腺癌、慢性萎缩性胃炎和浅表性胃炎新鲜组织中pERK mRNA表达;免疫组化法分别检测胃腺癌、慢性萎缩性胃炎和浅表性胃炎组织中pERK蛋白表达,并分析其蛋白表达与胃癌临床病理参数间的相关性。[结果]RT-PCR半定量结果显示,胃癌组织中pERK mRNA的相对表达水平（2.35±0.36）明显高于慢性萎缩性胃炎组织（1.18±0.25）及浅表性胃炎组织（0.68±0.10）（P均〈0.01）。免疫组织化学结果显示,pERK蛋白在胃癌组织中的阳性表达率（88.3%）高于慢性萎缩性胃炎（43.3%）及浅表性胃炎组织（5.0%）（P均〈0.01）。胃癌组织中pERK蛋白表达与胃癌分化程度、分期、淋巴结转移等明显相关。[结论]pERK在胃癌中高表达,pERK在正常细胞向恶性细胞转化的过程中可能扮演了重要角色,检测pERK表达可能有助于胃腺癌的预防及早期诊断。     

石蜡包埋的肝细胞癌组织中ASPP家族的表达

目的：研究肝细胞癌中p53凋亡刺激蛋白（ASPP）家族的表达以及临床意义。方法：应用免疫组织化学SP方法检测73例甲醛固定和石蜡包埋的肝细胞癌组织切片和相应癌旁正常组织中ASPP蛋白家族的表达情况。结果：在癌旁正常组织中,ASPP1和ASPP2的表达阳性率高于肝细胞癌组织（P=0．000,0．000）,但是肝细胞癌组织中iASPP的表达阳性率高于相应癌旁正常组织（P=0．000）。在肝细胞癌组织中,ASPP1的表达与患者性别、年龄、临床分期以及淋巴结转移均无相关性（P〉0．05）,但与病理分级有关（P=0．001）;ASPP2的表达与病理分级和临床分期有关（P=0．000,0．000）,与胜别、年龄以及淋巴结转移无关（P〉0．05）。iASPP的表达与淋巴结转移有关（P=0．004）,但是与性别、年龄、病理分级和临床分期无关（P〉0．05）。结论：ASPP蛋白家族异常表达可能与肝细胞癌的发生机制相关,ASPP1和ASPP2表达水平下调并与肝细胞癌病理分级相关,而iASPP的表达水平上调与癌的淋巴转移相关,iASPP可能是基因治疗肝细胞癌的一个靶点。     

肝细胞癌凋亡、p21和PCNA表达与临床预后关系的研究

目的：研究ＨＣＣ细胞凋亡,ｐ２１和ＰＣＮＡ表达与临床预后的关系。方法;５５例ＨＣＣ标本采用ＤＡＮ末端（Ｔｕｎｅｌ）和免疫化检测并结合计算机图像技术定量分析。结果：小肝癌ＡＩ高于大肝癌,肝内转移,浸润性及低分化ＨＣＣ的ＡＩ分别低于无转移,非浸润性及高分化ＨＣＣ。而ＰＣＮＡ－ＬＩ恰好相反。癌旁组织ｐ２１阳性率及ｐ２１表达值均高于癌组织。并且癌及癌旁组织ｐ２１表达值在浸润性及肝内转移ＨＣＣ分析高于非侵蚀     

MMP-2 mRNA及蛋白的表达在原发性 肝细胞癌组织中的意义

 目的 探讨原发性肝细胞癌（HCC）组织中基质金属蛋白酶-2（MMP-2）mRNA和MMP-2蛋白的表达,及其与临床病理参数之间的关系。方法 应用RT-PCR技术、免疫组织化学Maxvison^TM法,分别检测47例HCC组织及其癌旁组织、20例正常肝组织中MMP-2 mRNA和MMP-2蛋白的表达情况。结果（1）MMP-2 mRNA和MMP-2蛋白在肝癌中的表达分别为70.2%（33/47）和63.8％（30/47）,均高于癌旁组织12.7％（6/47）和正常组织10％（2/20）, 差异有统计学意义(P＜0.01), 癌旁组织和正常组织中MMP-2的表达差异无统计学意义(P>0.05);MMP-2mRNA和MMP2蛋白在肝癌中表达的两种检测方法差异无统计学意义(P>0.05);（2）癌组织中MMP-2的表达在TNM分期Ⅲ～Ⅳ期组中的阳性表达率高于Ⅰ～Ⅱ期组(P＜0.05),血管内有肿瘤栓组MMP 2阳性率明显高于无瘤栓组(P＜0.05）,有转移组的阳性率高于无转移组(P＜0.05）。结论 MMP-2的表达与HCC的TNM分期、血管内有无瘤栓和转移密切相关,可作为判断HCC侵袭性的评估指标之一。