肝细胞癌中miR-122的表达及其与血清AFP水平的关系

目的 探讨miR-122在肝细胞癌（肝癌）中的表达水平及其与血清AFP水平的关系。方法 用荧光定量PCR分别检测78例肝癌患者肝癌组织中miR-122的表达水平,电化学发光免疫分析法检测肝癌患者术前血清AFP水平,分析miR-122表达与血清AFP水平的关系。结果78例肝癌患者肿瘤组织miR-122的相对表达量为（23.71±13.33）％,其中高、中分化组织为（26.90±13.64）％,明显高于低分化者的（16.54±9.40）％（P=0.001）。78例肝癌患者血清AFP平均值为（741±382）ng／ml。miR-122表达与性别、年龄、淋巴结转移、肿瘤大小、血管浸润、分期等无关,miR-122在肝癌和癌旁组织中的表达与血清AFP水平呈负相关（r=-0.863,P＜0.01）。结论 miR-122表达与血清AFP水平呈负相关,在低分化肝癌中表达更低,提示miR-122是肝癌的一个潜在预后因子。     

以AFP为靶点的肝癌树突状细胞免疫治疗的实验研究

目的探讨AFP作为肝癌基因治疗和免疫治疗靶点的可行性以及AFP基因转染的树突状细胞（AFP-DC）疫苗对表达AFP肝细胞癌的免疫治疗作用.方法构建AFP-cDNA 真核表达载体,体外转染DC,制备AFP-DC瘤苗,诱导针对表达AFP的肝癌细胞株HepG2的特异性免疫反应,MTT法检测效应细胞对靶细胞的杀伤率.结果 AFP-DC瘤苗能够分泌AFP抗原,培养上清中AFP含量为0.8805 IU/ml,与空载体组和空白对照组相比,差异显著（P〈0.05）.免疫荧光检测可见AFP-DC胞浆及胞膜有AFP抗原分子表达;活化的CTL能够对表达AFP肝癌细胞起特异性杀伤作用,杀伤效率可达84.05%,与空载体组和空白对照组比较,差异显著（P〈0.05）.结论 AFP作为靶点治疗肝癌具有可行性,AFP可作为肝癌靶向治疗的新的突破点;AFP-DC疫苗可以作为表达AFP肝癌的一种免疫治疗手段,为DC瘤苗的临床应用奠定了基础.     

Hereditary persistence of {alpha}-fetoprotein: Case report and review of the literature

Persistently elevated -fetoprotein (AFP) levels of 24 to 30µg/ml (normal <10 µg/ml) were found in a 38-year-old healthyman. Subsequently, AFP was found to be elevated in another five out of 13family members within three generations. The pedigree is consistent with anautosomal dominant inheritance pattern. No discernible disease and nofunctional abnormality appears to be associated with this clinically benigndisorder which has been recorded in the literature on four occasions todate. The reported AFP levels in these other cases ranged from 18 to 198µg/ml.Physiologically, AFP is mainly produced in the liver and the yolk sac ofhuman fetuses more than four weeks old, with peak values of up to 4 mg/ml at12 to 16 weeks of gestation. After birth, AFP levels usually fall, withineight to 12 months, to a very low concentration of <10 µg/ml andpersist at low levels throughout life. However, AFP levels can rise abovenormal in both children and adults in distinct conditions and diseases whichwill be discussed.Hereditary persistence of -fetoprotein (HPAFP) should be consideredin both children and adults with unexplained and persistent elevation of AFPe.g., those screened for hepatocellular carcinoma or diagnosed for germ celltumor. It should also be recognized in AFP screening for neural tube defectsor Down's syndrome during pregnancy. Hereditary persistence of AFP can beeasily confirmed by analyzing AFP levels in family members.     

Autoantibodies to tumor-associated antigens combined with abnormal alpha-fetoprotein enhance immunodiagnosis of hepatocellular carcinoma

The identification and characterization of tumor-associated antigens (TAAs) and their use in antigen mini-arrays for cancer immunodiagnosis has been of interest recently as an approach to cancer detection. In this study, autoantibodies in sera from a patient with HCC were used as probes to immunoscreen a HepG2 cDNA expression library for the identification of TAAs involved in malignant liver transformation. Recombinant proteins from two genes identified in this manner, Sui1 and RalA were expressed, purified and used as antigens in immunoassays to detect the presence of antibodies in sera from 77 patients with HCC, 30 with chronic hepatitis (CH), 30 with liver cirrhosis (LC) and 82 normal human sera (NHS). The prevalence of antibody to Sui1 and RalA in HCC were 11.7% (9/77) and 19.5% (15/77), respectively, which were significantly higher than prevalence in liver cirrhosis (3.3% and 3.3%), chronic hepatitis (0% and 0%) and normal human sera (0% and 0%). When Sui1 and RalA were added to a panel of eight other TAAs used in a previous study, the final cumulative prevalence of anti-TAA antibodies in HCC to the 10 TAA array was raised to 66.2% (51/77). The specificity for HCC compared with LC, CH and NHS, was 66.7%, 80.0%, and 87.8%, respectively. When anti-TAA was added to abnormal serum AFP as combined diagnostic markers, it raised the diagnostic sensitivity from 66.2% to 88.7%. AFP and anti-TAA were independent markers and the simultaneous use of these two markers significantly resulted in the increased sensitivity of HCC detection.     

热休克蛋白72-甲胎蛋白抗原表位肽复合物抗小鼠肝癌Hepal-6细胞免疫的实验研究

目的:以热休克蛋白72(HSP72)-甲胎蛋白(AFP)抗原表位肽复合物免疫小鼠,研究该复合物针对AFP肿瘤是否具有特异性抗肿瘤免疫.方法:以HSP72-AFP多肽复合物皮下注射免疫昆明小鼠,并分别以AFP多肽和HSP72单独免疫小鼠为对照组.ELISA法检测免疫后小鼠血清IFN-γ水平、MTT法检测各免疫组小鼠淋巴细胞对肝癌Hepal-6细胞的杀伤作用、以小鼠体内瘤负荷实验评价蛋白复合物的免疫效应.结果:HSP72-AFP多肽复合物组小鼠血清IFN-γ水平、淋巴细胞对Hepal-6细胞的杀伤作用均显著高于AFP多肽和HSP72组(P＜0.01).HSP72-AFP多肽复合物组瘤体积也明显小于AFP多肽和HSP72免疫组(P＜0.01).结论:HSP72-AFP多肽复合物疫苗可诱导荷瘤小鼠产生针对AFP肿瘤的特异性细胞免疫,其对瘤细胞的杀伤效应明显优于两者单一的多肽纯化疫苗.表明HSP72-AFP多肽复合物可诱导小鼠产生有效的抗肿瘤免疫.     

Silencing alpha-fetoprotein expression induces growth arrest and apoptosis in human hepatocellular cancer cell

The expression of alpha-fetoprotein (AFP), a tumor-associated antigen, is silenced in normal adult hepatocyte but reactivated in human hepatocellular carcinoma (HCC). To investigate the roles of AFP in the regulation of cell growth, we silenced AFP expression in the HCC cell line Huh7 by transfection of specific Stealth RNAi. After the transfection for 48 h, the expression of AFP gene was almost abolished, the cell proliferation was inhibited by 46.15%, and the number of cells undergoing early apoptosis was significantly increased to 63.93%. Inhibition of AFP expression also resulted in an increased in Bax/Bcl-2 ratio, the release of cytochrome c from mitochondria and activation of caspase-3. The results suggest that AFP may positively regulate cell proliferation by enhancing the apoptosis resistance via dysfunction of the p53/Bax/cytochrome c/caspase-3 signaling pathway in AFP-producing HCC cell line. As such, the knockdown of AFP gene should be further investigated in vivo as a novel approach to HCC treatment.     

肝癌患者74例免疫球蛋白、补体水平及AFP分析

目的　探讨肝癌患者体液免疫指标血清球蛋白、IgG、IgA、IgM及补体 3 (C3 )、补体 4(C4)的变化及其与AFP的关系。方法　用自动生化分析仪和放免法定量测定肝癌患者的血清球蛋白、IgG、IgA、IgM、C3、C4及AFP的含量 ,设肝良性疾病肝硬化和正常人两组为对照组 ,运用SPSS软件进行统计。结果　肝癌和肝硬化组的球蛋白、IgG、IgA比正常人明显升高 (P <0 .0 1) ;肝癌组C3比正常人低 (P <0 .0 1) ,C4则高于正常人 (P <0 .0 1) ;AFP阳性和AFP阴性肝癌患者的免疫球蛋白、补体均无差异 (P >0 .0 5 ) ;肝癌患者的AFP与免疫球蛋白、补体无相关性。结论　肝癌患者的体液免疫功能增强 ,AFP与免疫球蛋白、补体无相关性。     

miR-1236 down-regulates alpha-fetoprotein,thus causing PTEN accumulation,which inhibits the PI3K/Akt pathway and malignant phenotype in hepatoma cells

Alpha fetoprotein (AFP) is a clinical biomarker of hepatocellular carcinoma (HCC). Here, we found that miR-1236 is down-regulated, whereas AFP is highly expressed in HCC tissues and cells. We demonstrated that miR-1236 directly targets the 3′UTR of AFP and down-regulates its expression. Also, miR-1236 inhibited and AFP stimulated proliferation, migration, invasion and vasculogenic mimicry (VM) of HCC. In agreement, AFP over-expression counteracted the inhibitory effect of miR-1236. We demonstrated that AFP promoted the ubiquitination of PTEN, thus decreasing PTEN levels, while miR-1236 inhibited the PI3K/Akt pathway.     

Hereditary alterations in tumor-host relationship

An indirect exposure of mice during their fetal stage of development to Ehrlich ascites tumor cells alters the animals' response to this tumor during their adult life. Control mice injected intraperitoneally with Ehrlich tumor cells react in various ways to the presence of this alien tissue. The response is expecially evident during the first 48–72 h post-injection. Several aspects of this complex reaction are absent in those animals which were exposed to this tumor in utero. The measurably altered reactivity toward the Ehrlich tumor cells is hereditary. Possible mechanisms of this alteration are discussed.     

多项指标联合检测诊断原发性肝癌的研究

目的 :探讨五项肿瘤标志物联检对提高原发性肝癌 (PHC)的检出率和早期诊断的意义。方法 :以甲胎蛋白 (AFP)的检测为主体配合谷氨酰转肽酶 (γ GT)、α L 岩藻糖苷酶 (AFU)、肿瘤坏死因子 (TNF α)和DR 70 TM五项指标对原发性肝癌 50例、肝外恶性肿瘤和肝硬化及健康人各 30例进行了血清检测。结果 :原发性肝癌患者多项指标含量与正常组比较均有明显差异或非常显著差异 (P <0 0 5和P <0 0 1)。各项指标除DR 70 TM和TNF α标志物对肝外恶性肿瘤和肝硬化比较差异无显著性外 ,其余指标各组间比较均有显著或非常显著差异 (P <0 0 5和P <0 0 1)。结论 :应用多项指标联合检测PHC的诊断率可达 98% ,尤其可提高小肝癌 (直径≤ 5cm)的检出率     

纵隔卵黄囊瘤并发脑转移1例并文献复习

卵黄囊瘤(yolk sac tumor,YST),又名内胚窦瘤(endodermal sinus tumor),是一种起源于生殖细胞的高度恶性肿瘤。大约占原始生殖细胞肿瘤的20%。纵隔卵黄囊瘤的发生,是由于生殖细胞从卵黄囊向生殖嵴移行过程中,被阻留在纵隔处或异常移居的结果。发生在性腺外者,因罕见或对其认识不足易误诊,本文报告1例患者的诊治过程,并就其临床病理形态学特点及其组织起源等问题,结合复习文献加以分析。     

Clinical Value of Hepatoma-Specific Alpha-Fetoprotein in the Diagnosis of Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) is a common malignant tumor worldwide and the number 2 cause of cancer mortality in Chi- na.[1] It often develops in cases of liver cirrhosis and chronic hepati- tis. [1-3] Advanced imaging procedures including utrasonograph…     

胃肝样腺癌伴肝转移治疗成功1例报告

我们报告一例成功治疗胃肝样腺癌(hepatoid adenocarcinoma of stomach,HAS)伴肝转移的病例,并复习了相关文献。患者男性,60岁,诊断为HAS伴肝转移,血AFP 780ng/ml,行根治性胃大部切除和肝切除术。术后经肝动脉行灌注化疗后,血AFP降至正常。胃的肝样腺癌(HAS)是具有肝细胞样分化和腺癌分化的胃癌,通常可以产生甲胎蛋白(AFP),易早期发生肝脏转移,预后不良,平均生存期<1年。此例病人随访8年,无肿瘤复发迹象。提示对HAS伴肝转移的病人仍有积极治疗的价值。     

Targeting gene therapy for hepatocarcinoma cells with the E. coli purine nucleoside phosphorylase suicide gene system directed by a chimeric alpha-fetoprotein promoter

For hepatocarcinoma (HCC) gene therapy, the tumoricidal efficacy and selective expression of therapeutic gene remain two major challenges. The Escherichia coli (E. coli) purine nucleoside phosphorylase (PNP)/9-(2-deoxy-beta-dribofuranosyl)-6-methylpurine (MeP-dR) suicide gene system exhibits excellent anti-tumor effects, indicating this system directed by a HCC-specific promoter would offer a possibility of targeting gene therapy for HCC. To test this hypothesis, here, we prepared a plasmid (p[HRE]AF/PNP) containing the E. coli PNP/MeP-dR system and a chimeric human alpha-fetoprotein (AFP) promoter, [HRE]AF. We introduced this plasmid into AFP-positive and low-AFP-generating human HCC cells, and evaluated its therapeutic effects on both human HCC cell lines. In the presence of hypoxia, the E. coli PNP gene directed by the [HRE]AF promoter were HCC-specifically expressed in two human HCC cell lines and, moreover, the [HRE]AF-PNP/MeP-dR therapy would yield significant and selective cytotoxicity in both AFP-positive and low-AFP-generating HCC cells. Our findings suggest the [HRE]AF-PNP/MeP-dR therapy has worthy potentialities as an effective strategy for targeting therapy of AFP-positive, and especially AFP-negative or low-AFP-generating HCC.     

含人AFP基因重组腺病毒载体的构建及其转染树突状细胞瘤苗的制备

目的:构建含人AFP基因的腺病毒载体,体外转染树突状细胞,制备树突状细胞肝癌瘤苗.方法: 将AFP基因亚克隆到pIND 载体和Shuttle2载体中,构建穿梭载体Shuttle2-AFP.用PI-Sce Ⅰ和I-CeuⅠ双酶切后将所获AFP基因片段再与线性化的腺病毒载体pAdeno-X连接,构成pAdeno-AFP重组腺病毒载体.其后,用重组腺病毒载体转染HEK293细胞,包装腺病毒表达载体.通过酶切、PCR对腺病毒载体进行鉴定.包装好的重组病毒载体pAdeno-AFP体外感染树突状细胞制备树突状细胞肝癌瘤苗后,FACS分析pAdeno-AFP/DC表面分子的表达,酶联免疫吸附试验法( ELISA) 检测AFP水平.结果: 酶切、PCR鉴定证实,穿梭质粒插入片段为AFP基因.包装的腺病毒载体具有良好的感染性,可以在293细胞中形成病毒颗粒,腺病毒载体内携带AFP基因感染树突状细胞,pAdeno-AFP/DC能高水平的表达CD1a,CD11c,CD80,CD86以及HLA-DR,并分泌较高水平的AFP.结论: 构建成功的含AFP腺病毒载体可以在树突状细胞中表达AFP,为DC瘤苗的进一步研究奠定了基础.     

Immunohistochemical Type Distinction of a-Fetoprotein in Various a-Fetoprotein-secreting Tumors

In order to clarify the histogenesis of a -fetoprotein(AFP)-secreting tumor tissues, formalin-fixed, paraffin-embedded serial sections of 148 tumors in various organs were examined by the peroxidase-antiperoxidase method for AFP, and paradoxical concanavalin A staining. Yolk sac-type AFP was found in yolk sac tumors, embryonal carcinomas, solid teratomas, (yolk sac) endodermal cell tumors, adenocarcinomas (stomach, ovary or lung) and metastatic liver cancers. Hepatic-type AFP was demonstrated in hepatocellular carcinomas, hepatoblastomas, solid teratomas and a stomach cancer. Yolk sac-type AFP was observed in the neighboring liver cells of metastatic liver cancers without relation to the type of AFP in primary cancers. The results from serum analyses of preoperative tumor-bearing patients (68 cases) were coincident with those from immunohistochemical stainings.     

家族性胃癌研究进展

The etiology of familial gastric cancer (FGC) are E-cadherin (CDH1) mutations, helicobacter pylori infection, etc. The median age of patients with FGC is 50-65 years old. The male-female sex ratio is 2:1. The most of tumors of FGC locate in distal stomach and present diffuse type in histological classification. It is reported that the people who has a family history of gastric cancer especially for those parents has an increased risk of gastric cancer. It is difficult to find FGC in early stage by current technology. The preventive methods include eradication of helicobacter pylori and prophylactic total gastrectomy, etc.     

Family cancer history and risk of brain tumors in children: results of the SEARCH international brain tumor study

OBJECTIVE: To examine whether childhood brain tumors (CBTs) are associated with a family history of brain tumors or other cancers in an international case-control study. METHODS: Cancers in children's first- and second-degree relatives were ascertained by interview with parents of 620 children with astroglial tumors, 255 with primitive neuroectodermal tumors, 324 with other CBTs, and 2,218 controls from Australia, Canada, France, Israel, Italy, Spain, and the US. These were used with histories of neurofibromatosis or tuberous sclerosis to exclude in subanalyses children with Li-Fraumeni or other hereditary syndromes predisposing to brain tumors. RESULTS: A first- or second-degree relative of 4% of children with astroglial tumors, 6% with PNET, 5% with other CBTs, and 5% of controls had had a brain tumor. Any potential differences were statistically non-significant, including when focusing on relatives diagnosed in childhood. In the US, where anatomical sites of relatives' other cancers were known, CBT occurrence was not associated with any other specific site. Results were not markedly altered by exclusion of children with hereditary syndromes. CONCLUSION: Consistent with most prior studies using these methods, we observed no strong relationship between CBT occurrence and cancers in family members.