Studies on mechanism of Sialy Lewis-X antigen in liver metastases of human colorectal carcinoma

INTRODUCTIONSialyl Lewis-X antigen ,correlated with carcinoma, is a group of carbohydrate antigen containing oligosaccharide expressed of embryonic tisue and glycoproteins on cell surface of embryonic tissue[1].The SLeX antigen located on cell surface is synthesized principally by two enzymes ,al ,3fucosyltransfrease and a2, 3sialyctransferase.In adults ,SLeX antigen is expressed principally on the surfaces of granulocytic cells and some tumor cells .

Relationship between phenotypes of cell-function differentiation and pathobiological behavior of gastric carcinomas

AIM: To reveal the correlation between the functional differentiation phenotypes of gastric carcinoma cells and the invasion and metastasis by a new way of cell-function classification.METHODS:Surgically resected specimens of 361 gastric carcinomas(GC) were investigated with enzyme-, mucin-, and tumor-related marker immunohistochemistry. According to the direction of cell-function differentiation, stomach carcinomas were divided into five functionally differentiated types. RESULTS: (1) Absorptive function differentiation type (AFDT): there were 82 (22.7%) patients including 76 (92.7%) aged 45 years. Sixty-nine (84.1%) cases belonged to the intestinal type. Thirty-eight (46.3%) expressed CD44v6 and 9 (13.6%) of 66 male patients developed liver metastasis.The 5-year survival rate of patients in this group (58.5%) was higher than those with the other types (P【0.01). (2) Mucin secreting function differentiation type (MSFDT): 54 (15%) cases. Fifty-three (98.1%) tumors had penetrated the serosa, 12 (22.2%) expressed ER and 22 (40.7%) expressed CD44v6. The postoperative 5-year survival rate was 28.6%. (3) Absorptive and mucin-producing function differentiation type (AMPFDT): there were 180 (49.9%) cases, including 31 (17.2%) aged younger than 45 years. The tumor was more common in women (62, 34.4%,) and expressed more frequently estrogen receptors (ER) (129, 81.7%) than other types (P【0.01). Ovary metastasis was found in 12 (19.4%) out of 62 female subjects. The patients with this type GC had the lowest 5-year survival rate (24.7%) among all types. (4) Specific function differentiation type (SFDT): 13 (3.6%) cases. Nine (69.2%) tumors of this type derived from APUD system, the other 4 (30.7%) were of different histological differentiation. Sixty per cent of the patients survived at least five years. (5) Non-function differentiation type (NFDT): 32 (8.9%) cases. Nineteen (59.4%) cases had lymph node metastases but no one with liver or ovary metastasis. The 5-year survival rate was 28.1%. CONCLUSION: This new cell-function classification of GC is helpful in indicating the characteristics of invasion and metastasis of GC with different cell-function differentiation phenotypes. Further study is needed to disclose the correlation between the cell-functional differentiation phenotypes and the relevant genotypes and the biological behavior of gastric carcinoma.

Expressions of ICAM-1 and its mRNA in sera and tissues of patients with hepatocellular carcinoma

INTRODUCTIONThe increased expression of ICAM-1 on a widerange of cells and in the sera of patients withmalignancies, chronic liver diseases andinflammation diseases has been described since thelate 1980s[1-22]. Recently rapid progress in studieson expression of ICAM-1 in patients withhepatocellular carcinoma ( HCC ) have beenachieved, including clinical and experimentalresearches[23-31].

补肾方对大鼠肝癌PCNA、p53蛋白的影响

目的研究补肾方对二乙基亚硝胺( DEN)诱导大鼠肝癌过程中增殖细胞核抗原( PCNA)、( P5 3)蛋白表达的影响。方法建立DEN诱导大鼠肝癌模型,分别于诱癌第1 2周末和第1 8周末采用免疫组化染色法检测补肾方治疗前后大鼠肝细胞PCNA、P5 3蛋白。结果P5 3阳性细胞可见于癌前肝细胞的增生结节中;补肾方在诱癌第1 2周末和第1 8周末均能明显抑制P5 3的阳性表达( P<0 .0 5 ) ,显著降低PCNA- LI( P<0 .0 1 )。结论在二乙基亚硝胺( DEN)诱导的大鼠肝癌模型中P5 3基因突变可发生于肝癌癌前病变,参与肝癌的启动过程。补肾方可抑制大鼠诱癌过程中肝细胞的增殖活性和P5 3的突变,从而延缓肝癌的形成。

Immunohistochemical study on p53,H-rasp21,c-erbB-2 protein and PCNA expression in HCC tissues of Hart and minority ethnic patients

AIM To find out the difference of humanprimary liver carcinogenesis between Han andminority ethnic patients in Xinjiang.METHODS Expression of p53,c-erbB-2,H-rasp21 protein and proliferating cell nuclearantigen(PCNA)in tumor tissues of 50 patients(Han 38,minority 12)with primary hepaticcarcinoma was detected byimmunohistochemistry(LSAB).RESULTS The positive frequency of p53,c-erbB-2,H-rasp21 and PCNA expression was46.0%(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 peritumors respectively and asignificant difference,except for H-rasp21,ofoncogene alteration was found(P【0.05)between tumor and non-tumorous tissues.Combined the three oncogenes alteration,26%(13/50)tumor tissues had positiveimmunoreactivity,but in peritumor and normallivers it was negative.The positive rate of p53,c-erbB-2 and H-rasp21 protein expression was39.5%(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 minoritiesrespectively,with statistical difference (P【0.05).CONCLUSION Overexpression of p53,c-erbB-2and H-rasp21 in human primary liver carcinoma isan important biomarker of genetic alteration.The different frequency of these oncogeneticchanges may reflect some environmental or/andethnic hereditary factors affecting the livercarcinogenesis.The special life style of Han,Uygur,Kazak and Mongolia nationalities inXinjiang may also be related to theetiopathogenesis of this disease.

THE PHENOTYPE OF MAST CELL IN PRIMARY ADENOID LIVER TUMOR OF RAT AND ITS RELATION TO TUMOR CELL

Mast cells in adenoid liver tumors of 32 rats induced with nitrosomorpholine were observed ultrastructurally, and among them, some were studied immunocytochemically via immunogold techniques. Data indicating that mast cells which located in tumor tissues presented positive expression of rat mast cell protein (RMCP) Ⅰ, Indicating origination from the mucosa mast cells, while those in the connective tissues around tumors were largely stained negatively with either RMCP Ⅰor RMCP Ⅰ antisera, with the exception of only a few cells showing positive RMCP Ⅰ staining. Ultrastructural observation showed that mast cells in tumon contacted closely with the tumor cells. Membranes of the intracytoplasmic granules in these mast cells were fusing together. The content inside the granules were discharged and spread along the intercellular space between the tumor cells. There was not any lesion observed uitrasructrually in the tumor cells contacting with the mast cells. The significance of mucosa mast cells in adenoid liver tumors is briefly discussed.

阿托伐他汀抑制肝癌细胞增殖作用的研究

目的:体外观察阿托伐他汀对人肝癌HepG2细胞增殖、细胞周期及COX-2蛋白表达的影响。方法:取对数期人肝癌HepG2细胞,加入阿托伐他汀使其终浓度为0、0.1、1、10和100μmol/L,采用细胞计数法及四甲基偶氮唑盐(MTT)法观察阿托伐他汀对HepG2细胞增殖的影响,流式细胞仪(FCM)研究阿托伐他汀对细胞周期的作用,免疫细胞化学观察阿托伐他汀对COX-2蛋白表达的影响。结果:体外阿托伐他汀呈浓度和时间依赖性地抑制HepG2细胞的增殖,但0.1μmol/L组与0 μmol/L组差异无统计学意义。细胞周期分析显示,阿托伐他汀作用24h后,呈浓度依赖性改变细胞周期分布,G0/G1期细胞的比例增高,S期及G2/M期的细胞比例降低,但细胞凋亡不明显。体外阿托伐他汀呈浓度依赖性地抑制HepG2细胞COX-2蛋白的表达。结论:体外阿托伐他汀对HepG2细胞增殖有抑制作用,该作用可能与使细胞生长阻滞于G0/G1期及抑制COX-2蛋白表达有关。

不同斑蝥素类物质对人肝癌细胞HepG_2增殖作用的研究

目的观察斑蝥素酸镁、斑蝥素及斑蝥素酸钠3种斑蝥素类物质对人肝癌细胞Hep G2的体外抗肿瘤活性。方法 (1)采用磺酰罗丹明染色法(SRB法)检测不同浓度斑蝥素酸镁、斑蝥素及斑蝥素酸钠在体外对人肝癌细胞Hep G2增殖的抑制作用;(2)使用透射电镜观察2.27μmol/L斑蝥素酸镁作用于肝癌细胞Hep G248 h后细胞超显微结构的变化。结果 (1)斑蝥素酸镁、斑蝥素及斑蝥素酸钠对人肝癌细胞Hep G2有明显抑制作用,抑制率随药物浓度的增加而升高,且呈剂量效应关系,其半数抑制浓度(IC50)分别为2.27、5.70、8.41μmol/L;(2)透射电镜下见核染色质聚集成块附着于核膜下,且见核异形等凋亡细胞形态特征。结论斑蝥素酸镁对人肝癌细胞Hep G2的抑制作用优于斑蝥素和斑蝥素酸钠,且斑蝥素酸镁可以诱导人肝癌细胞Hep G2发生凋亡。

Reversing effect of Tanshinone on malignant phenotypes of human hepatocarcinoma cell line

AIM To study the reversing effect of Chinese drug tanshinone on malignant phenotype of cancer cells.METHODS Human hepatocarcinoma cell line (SMMC-7721) was treated in vitro with 0.5mg/L tanshinone for 4 days, and variation in cell differentiation was detected.RESULTS The morphology of cancer cells was tended toward well differentiation and cell growth was markedly inhibited. BrdU uptake assay and immunohistochemical stain of PCNA showed that the BrdU labeling rate and PCNA positive rate were lower than the controls, but no difference was found statistically as compared with all transretinoic acid. Flow cytometric assay demonstrated that S phase cells decreased and G0/G1 phase cells increased. Expression of c-myc oncogene protein decreased but the c-fos oncogene protein markedly increased.CONCLUSION Tanshinone could reverse the inducing differentiation in human hepatocarcinoma cells (SMMC-7721). It may become a new prospective inducer of cell differentiation to treat cancers.

肝细胞癌中血管生成拟态的三维细胞培养及组织学研究

背景与目的:血管生成拟态(vasculogenic mimicry,VM)是新近发现的一种存在于恶性肿瘤中的血液供应方式。拟态血管由肿瘤细胞围成,无内皮细胞衬覆的管道样结构,其间有血液通过。目前对拟态血管的研究多局限在高侵袭性、双向分化的恶性肿瘤,对上皮来源的肿瘤很少涉及。本研究通过细胞三维培养,免疫组织化学方法观察肝细胞癌(hepatocellular carcinoma,HCC)中是否存在血管生成拟态,并探讨其形成过程及结构特点。方法:建立肝癌细胞株HepG2的三维培养模型,并收集15例肝癌石蜡包埋样本进行过碘酸雪夫氏(PAS)及CD31免疫组织化学双重染色以及PAS与Ferritin免疫组织化学双重染色,观察肝细胞肝癌中的血管生成拟态。结果:肝癌细胞三维培养两天时伸出细长突起,培养7天时彼此之间相互连接形成网络样、环状结构。15例肝癌免疫组织化学染色可见血管内皮CD31染色均为阳性,肝癌细胞Ferritin染色均为阳性。其中7例免疫组织化学与组织化学双重染色可见CD31阴性Ferritin阳性的肝癌细胞围成管道样结构,一层PAS阳性物质将肿瘤细胞同管腔分隔,肿瘤细胞构成的管腔中可见红细胞存在。并观察到分化良好的肝癌中血管生成拟态的形成少于分化差的肝癌,随着分化程度的降低肝癌细胞逐渐出现CD31着色阳性倾向。结论:肝癌细胞具有进行自身变形并与胶原蛋白相互作用,形成血管样通道的能力。肿瘤细胞可以通过血管生成拟态结构获得血液供应。

Expression of fibronectin and cell adhesion molecule CD15 in hepatocellular carcinoma

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肝细胞癌高表达中期因子与肝癌细胞血行播散的关系

目的:探讨肝细胞癌(HepatocellularCarcinoma,HCC)中期因子(Midkine,MK)蛋白高表达与肝癌细胞血行播散的可能关系。方法:应用RT-PCR和免疫组织化学染色方法分别检测AFP阳性HCC患者术前外周血AFPmRNA和手术切除肝癌组织MK蛋白,了解肝癌细胞血行播散和MK表达的情况,分析两者之间的相关性。结果:在41例HCC患者中,18例术前外周血可检出AFPmRNA,检出率为43.9%。在对应的肝癌组织中,29例高表达MK,表达率为70.7%。在29例高表达MK的HCC患者中,18例术前外周血AFPmRNA阳性,阳性率为62.1%;而在12例MK低表达的HCC患者中,仅3例术前外周血AFPmRNA阳性,阳性率为25.0%。两组之间差异有显著性(P<0.05)。结论:肝癌组织异常表达MK与外周血循环中存在肿瘤细胞有关,可能是肝癌细胞转移潜能的一个指标。

间隙连接Cx32 mRNA、Cx43 mRNA及其蛋白产物在肝细胞肝癌中的表达

目的：探讨间隙连接基因Ｃｘ３２ｍＲＮＡ、Ｃｘ４３ｍＲＮＡ及其蛋白产物在肝细胞癌（ｈｅｐａｔｏｃｅｌｕｌａｒｃａｒｃｉｎｏｍａ，ＨＣＣ）发生的重要意义。方法：应用Ｓ－Ｐ免疫组织化学法、原位杂交和流式细胞仪分析技术，研究６１例ＨＣＣ、１４例正常肝组织以及肝癌细胞系ＨＨＣＣ、ＳＭＭＣ－７７２１、正常肝细胞系ＱＺＧ中，Ｃｘ３２ｍＲＮＡ、Ｃｘ４３ｍＲＮＡ及其蛋白产物的表达规律。结果：Ｃｘ３２蛋白在ＨＣＣⅠ、Ⅱ、Ⅲ级和正常肝组织中的阳性率分别为５５．６％、４２．１％、１８．２％和９２．９％；Ｃｘ３２蛋白在ＨＨＣＣ、ＳＭＭＣ－７７２１和ＱＺＧ中阳性细胞计数率分别为１．９％、１．７％和９９．０％。Ｃｘ４３蛋白在ＨＣＣⅠ、Ⅱ、Ⅲ级和正常肝组织中的阳性率分别为４４．４％、２６．３％、１２．１％和７８．６％。Ｃｘ４３蛋白在ＨＨＣＣ、ＳＭＭＣ－７７２１和ＱＺＧ中阳性细胞计数率分别为７．３％、２．３％和９９．１％。Ｃｘ３２、Ｃｘ４３在ＨＣＣ各级和正常肝组织中差异有显著性（Ｐ＜０．０１），在肝癌细胞系ＨＨＣＣ、ＳＭＭＣ－７７２１和正常肝细胞系ＱＺＧ中阳性细胞计数率差异有显著性（Ｐ＜０．０１）。原位杂交显示，Ｃｘ３２ｍＲＮＡ在ＨＣＣⅠ、?

人肝癌与癌旁肝AFP阳性细胞形态的图象分析及增殖活性的研究

采用免疫组化、双标免疫组化及图象分析技术，研究了２７例人体肝细胞癌与癌旁肝甲胎蛋白（ＡＦＰ）阳性细胞的量化形态特征及以其增殖细胞核抗原（ＰＣＮＡ）标记率显示的增殖活性。主要结果显示：（１）ＡＦＰ阳性与阴性癌及癌旁近癌肝细胞间难以用量化形态标准区分（Ｐ＞０．０５）；核面积与核浆比提示癌旁ＡＦＰ阳性肝细胞可能是尚无表型转化的癌前细胞。（２）ＡＦＰ阳性癌细胞ＰＣＮＡ的平均标记率（１５．７０％±６．２５％）显著低于肝癌的平均标记率（Ｐ＜０．０１），表明大多数ＡＦＰ阳性癌细胞处于非增殖状态。（３）癌旁近癌肝可见ＡＦＰ与ＰＣＮＡ双标阳性肝细胞。

原发性肝脏弥漫大B细胞淋巴瘤7例临床病理学观察

目的 探讨肝脏原发性弥漫大B细胞淋巴瘤（primary hepatic diffuse large B-cell lymphoma,PHDLBL）的影像学、组织学及免疫表型,分析其临床病理特征、诊断、鉴别诊断、治疗及预后。方法 对7例肝脏PHDLBL进行光镜观察及免疫组化标记,采用原位分子杂交法检测淋巴瘤组织中EBV编码的小RNA（EBV-encoded small RNA,EBER）,并结合相关文献复习。结果 7例患者平均年龄67.5岁,均以右上腹部饱胀不适、消瘦伴贫血就诊。CT示病灶密度欠均匀,可见更低密度区,增强扫描病灶不均匀强化。肿瘤细胞弥漫性生长,瘤细胞体积较大,大小相对一致,多为圆形,胞质较丰富、淡染,核圆形或卵圆形,染色质粗,可见明显的核仁和泡状核,核分裂象易见,并可见星空现象。免疫表型：瘤细胞弥漫表达CD19、CD20、CD79a,CD3、CD43均阴性,Ki-67增殖指数75%~80%;3例EBER原位杂交检测呈阳性。结论 PHDLBL临床罕见,预后较差,肝活检及免疫组化标记有助于诊断,需与其他部位的淋巴瘤累及肝脏鉴别;及早手术和化疗有望获得较好疗效。

肝癌组织中DLK1和ABCG2的表达及其相关性的探讨

目的分析DLK1(delta drosophila homolog-like 1)和ABCG2(ATP-binding cassette superfamily G member2)在原发性肝细胞性肝癌(primary human hepatocellular carcinoma,HCC)中的表达及探讨两者之间的关系。方法分别应用免疫组织化学S-P法和原位杂交方法检测HCC150例、肝硬化50例、肝炎组织50例、正常肝组织10例中DLK1和ABCG2蛋白及mRNA的表达情况。结果 HCC、肝硬化、肝炎和正常肝组织的DLK1蛋白阳性率分别为64.7%、28.0%、6.0%、0.0%;DLK1 mRNA阳性率分别为55.3%、20.0%、10.0%、0.0%,DLK1蛋白在HCC中的表达与肿瘤大小、分化程度有关。ABCG2蛋白在HCC、肝硬化、肝炎和正常肝组织的表达率分别为78.7%、40.0%、60.0%、30.0%;ABCG2 mRNA阳性率分别为69.3%、36.0%、50.0%、20.0%,ABCG2蛋白在HCC中的表达与有无转移有关。DLK1、ABCG2在肝癌中的表达呈正相关。结论 DLK1、ABCG2的表达一定程度上反映了HCC侵袭性的强弱,在HCC的发生、发展过程中起着重要的作用。

PCNA在接受过术前经导管肝动脉栓塞化疗的肝癌组织中的表达及其预后意义

背景与目的:增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)可反映肝细胞癌(以下简称"肝癌")的增殖活性,经导管肝动脉栓塞化疗(transcatheter arterial chemoembolization,TACE)对术后残癌细胞增殖活性影响的报道结果不一。本研究旨在探讨术前TACE对肝癌增殖活性的影响和PCNA在二期切除肝癌复发预后中的意义。方法:选取91例术前TACE及50例未行TACE的肝癌手术切除标本,采用免疫组化方法检测两组标本PCNA的表达情况,分层分析两组PCNA表达的差异,以及术前TACE后二期根治性切除的肝癌中PCNA的表达水平与复发的关系。结果:PCNA表达于肝细胞核中,术前TACE组和未行TACE组的PCNA阳性率分别为67.0%和66.0%(P>0.05),术前TACE组中癌栓、播散结节或肿瘤低分化(Ⅲ～Ⅳ级)者PCNA的表达均明显低于对应分层的未行TACE者,差异有统计学意义(P<0.05)。术前TACE后根治性二期切除患者,复发组与未复发组肿瘤组织中PCNA阳性率分别为77.8%、47.1%,差异有统计学意义(P<0.05)。PCNA阳性组1、3、5年无瘤生存率分别为55.8%、31.8%、23.6%,PCNA阴性组分别为83.2%、62.1%、53.2%,差异有统计学意义(P<0.05)。结论:术前TACE对高侵袭肝癌细胞增殖活性有较好的抑制作用,术前TACE后二期根治性切除肝癌PCNA的表达与患者术后复发转移密切相关。

肝癌细胞及正常肝细胞中间隙连接蛋白Cx32、Cx43的表达——激光扫描共聚焦显微镜研究

目的 :研究间隙连接蛋白Cx32、Cx43在肝癌细胞系HHCC、SMMC 772 1及正常肝细胞系QZG中的表达。方法 :采用免疫荧光标记技术 ,对HHCC、SMMC 772 1及QZG细胞中Cx32、Cx43蛋白进行激光扫描共聚焦显微镜 (LSCM)分析。结果 :Cx32、Cx43蛋白定位于肝细胞胞质中 ,在HHCC、SMMC 772 1中阳性细胞率、阳性细胞的荧光强度较QZG中明显减弱。结论 :Cx32。

增殖细胞核抗原、胎盘型谷胱甘肽S-转移酶在肝细胞癌及癌周组织中的表达

目的探讨增生结节与肝细胞癌发生的关系。方法应用免疫组织化学方法，分别检测７０例肝细胞癌及癌周组织中增殖细胞核抗原（ＰＣＮＡ）的表达，以及７７例肝细胞癌和癌周组织中胎盘型谷胱甘肽Ｓ－转移酶（ＧＳＴπ）的表达。结果ＰＣＮＡ和ＧＳＴπ在肝细胞癌中的阳性表达率显著高于癌周组织，且ＰＣＮＡ与肝细胞癌Ｅｄｍｏｎｄｓｏｎ分级密切相关；癌周组织中，增生结节ＰＣＮＡ和ＧＳＴπ阳性表达显著高于肝硬变及正常组织。结论ＧＳＴπ可作为肝细胞癌前病变的标志酶；增生结节可能为肝细胞癌的癌前病变。

增殖细胞核抗原在原发性肝癌中的表达及其临床意义

目的 研究增殖细胞核抗原(PCNA)在原发性肝癌(PHC)中的表达及其与临床病理特征的关系。 方法 收集50例人PHC及其癌旁肝组织用标记链亲和素生物素(LSAB)法进行PCNA免疫组织化学研究。 结果 癌与癌旁肝组织PCNA标记指数(LI)有显著差异,癌的PCNALI平均为331±310,而癌旁为38±53,正常对照肝组织均阴性。癌组织分化越差,PCNALI越高,除Ⅲ与Ⅳ级外,PCNALI在组织学分级之间差异均具有显著意义。高表达PCNALI组(≥33%)在肝癌大小、组织学分级、肝硬化中的表达与低表达PCNALI组(<33%)之间差异均有非常显著意义。 结论 PCNALI是反映组织分化程度和生物学行为的一项重要指标。