NHE1 mRNA在肝癌组织的表达及意义

目的 探讨钠氢交换蛋白1（NHE1）mRNA在原发性肝细胞肝癌组织及癌旁组织的表达及意义。方法采用RT—PCR检测诊断为HCC患者34例的肝癌及癌旁组织中NHE1mRNA的表达情况。结果所有肝癌组织和癌旁组织NHE1均有表达,相对表达量分别为2．892±0．882和0．802±0．206,两者存在显著性差异（P〈0．001）。作为对照的21例肝组织NHE1的相对表达量为0．872±0．186。肝癌组织与对照肝组织比较存在显著性差异（P〈0．001）;癌旁组织与对照肝组织比较差异无统计学意义（P〉0．10）。将从同一患者取材的肝癌组织和癌旁组织检测到的NHE1mRNA的表达量进行配对t检验,两者间存在显著性差异（P〈0．001）。结论NHE1mRNA在肝癌组织的表达增高,可能在肝癌的发生中起一定的作用。     

巨噬细胞移动抑制因子和细胞周期蛋白D1的表达与肝细胞癌生长和转移的关系

目的研究巨噬细胞移动抑制因子（MIF）、细胞周期蛋白D1（Cyclin D1）、细胞周期蛋白依赖激酶4（CDK4）、磷酸化视网膜母细胞瘤易感基因产物蛋白（phospho—Rb）在HeC组织中的表达及其与癌细胞生长和转移的关系。方法应用组织芯片技术和免疫组织化学方法检测93份HCC组织中MIF、Cyclin D1、CDK4和phospho—Rb的表达，分析它们的表达与HCC临床病理特征的关系。结果93份HCC组织中MIF、Cyclin D1、CDK4和phospho—Rb的表达率分别为71％、41％、82％和14％，MIF和Cyclin D1阳性表达率与正常肝脏组织表达率之间的差异有统计学意义（P〈0．01），CDK4和phospho—Rb阳性表达率与正常肝脏组织表达率之间的差异无统计学意义（P〉0．05）。在≥3．5cm的肿瘤中MIF表达率为79％，明显高于在〈3．5cm肿瘤中的表达率48％（P〈0．01）；有转移的肝癌组织中Cyclin D1阳性率为62％，明显高于无转移组织中的阳性率35％，差异有统计学意义（P〈0．05）。MIF表达强弱与Cyclin D1的表达呈正相关关系（P〈0，01）。CDK4和phospho—Rb的表达与肿瘤大小及是否转移的关系无统计学意义。结论MIF和Cyclin D1在HCC发展进程中可能促进肿瘤的生长和转移。     

肿瘤特异性肿瘤/睾丸抗原在肝癌组织中的表达

目的 研究7种主要肿瘤/睾丸(CT)抗原MAGE-1、MAGE-3、MAGE-4、MAGE-10、NY-ESO-1、SSX-2、SCP-1在原发性肝细胞癌(HCC)患者癌组织中的表达、编码基因的变异状况及与临床指标的关系。 方法 收集30例肝癌患者的癌和癌旁组织,采用特异性引物逆转录聚合酶链反应检测7种CT抗原的表达,并对PCR产物进行测序分析。结果 在30例HCC患者中,MAGE-1、MAGE-3、MAGE-4、MAGE-10、NY-ESO-1、SSX-2、SCP-1在癌组织中的表达率分别为66.7％、70.0％、20.0％、36.7％、40.0％、33.3％和33.3％,而癌旁没有表达。癌组织中至少表达1种、2种和3种CT抗原的阳性率分别为90.0％、70.0％和53.5％。我国肝癌表达的7种CT抗原编码基因与国外报道的相比,同源性非常高。MAGE-10和SCP-1的表达与甲胎蛋白的水平相关,MAGE-3和SSX-2表达与平均年龄相关。 结论 7种CT抗原在HCC患者癌组织中均有表达,阳性率为20.0％-70.0％,其编码基因序列高度保守。一些CT抗原的表达与临床指标存在相关性。     

Glypican-3蛋白在肝细胞癌患者血清和组织中的表达及意义

目的:探讨肝细胞癌(HCC)患者血清和组织中Glypican-3(GPC3)蛋白的表达及临床意义.方法:收集HCC患者27例和肝良性病变患者28例的术前1d空腹血清及组织标本,采用Western blot检测血清中GPC3蛋白,同时采用免疫组织化学SP法检测组织标本中GPC3的表达情况.结果:27例HCC患者HCC组织、癌旁组织和远癌肝组织中GPC3蛋白阳性的表达率分别为81.5%,0%和0%(HC:23.4689,P<0.001),术前血清中GPC3蛋白阳性率分别为55.6%;28例肝良性病变患者组织和血清中阳性率均为0%;GPC3蛋白对HCC诊断的敏感性为55.6%,特异性为100%,误诊率为0%.HCC患者血清中GPC3蛋白表达与其瘤体大小和病理分级之间差异有显著性(P<0.05),而与患者年龄、性别、HBsAg及AFP值之间差异无显著性(P>0.05).结论:GPC3蛋白在HCC患者血清和组织中有较高的表达,对诊断HCC有较高的敏感性和特异性,可作为HCC早期诊断的标志物.     

Expression of the insulin-like growth factor system in skeletal muscle during embryonic and postnatal development in the first filial generation pigs from Erhualian and Yorkshire reciprocal crosses

In this study, we detected the expression of IGF-I, IGF-II, IGF-IR, IGF-IIR, and IGFBP-3 mRNA at 50 (E50), 70 (E70), and 90 (E90) days of gestation, and 1 (D1), 20 (D20), 70 (D70), 120 (D120), and 180 (D180) days of age in the longissimus dorsi (LD) and the semitendinosus (ST) of pigs from a Yorkshire boar × Erhualian sow (YE) cross as well as a Erhualian boar × Yorkshire sow (EY) cross. We found that the expression of IGF-I and IGF-II mRNA in skeletal muscle tissues differed based on developmental age and reciprocal cross type (P < 0.05). The expression of IGF-I mRNA exhibited a fluctuant ascending trend. In contrast, IGF-II showed a fluctuant descending trend after birth. The levels of IGF-IR mRNA were higher before birth compared with after birth except for the ST of EY pigs at D120 (P < 0.05). The expression of IGF-IIR and IGFBP-3 mRNA remarkably changed with age and reciprocal cross type (P < 0.05). IGF-I, IGF-II, and IGFBP-3 mRNA were positively correlated with IGF-IR from 50E to 180D. These data suggest that the expression of IGF-system genes exhibits specific developmental patterns in the skeletal muscle tissues of pigs from reciprocal crosses at different developmental stages and may show linked expression during certain periods of development. Our results may provide a valuable resource for the molecular breeding of pigs.     

MAGE-4基因在肝细胞肝癌中的表达及临床意义

目的通过研究MAGE-4基因在肝细胞肝癌组织中的表达,并与患者临床资料进行分析,探讨 MAGE-4基因与肝细胞肝癌(HCC)患者临床指标及转移与复发的关系,为MAGE-4基因编码蛋白用于HCC患者免疫治疗提供依据. 方法用RT-PCR的方法对31例HCC患者癌组织及相应癌旁组织MAGE-4基因表达进行测定,对全部RT-PCR扩增产物中目的基因片段进行DNA测序以证实其为 MAGE-4基因,患者均测定并统计AFP、AFU、抗HCV、HBsAg、AFP mRNA、肿瘤直径等临床指标. 结果 31例HCC患者肝癌组织中MAGE-4基因表达的阳性率38.7% (12/31)明显高于癌旁组织中MAGE-4基因表达的阳性率0% (0/31),P＜0.01.HCC患者肝癌组织中MAGE-4基因表达的阳性率与患者AFP、AFU、抗HCV、HBV标志物、AFP mRNA、肿瘤直径等临床指标均无关,P＞0.05. 结论 MAGE-4基因在HCC患者肝癌组织中特异高表达,可能作为HCC患者免疫治疗攻击的靶点,HCC患者肝癌组织中 MAGE-4基因表达的阳性率与HCC患者肿瘤标志物、转移、复发均无关.     

Tissue, developmental, and metabolic regulation of messenger ribonucleic acid encoding a rat insulin-like growth factor-binding protein

Insulin-like growth factor-II (IGF-II) is the predominant insulin-like growth factor in fetal and neonatal rat serum and tissues. In serum, it occurs complexed to a 30-kDa nonglycosylated IGF-binding protein (IGFBP) that is immunologically related to the IGFBP in BRL-3A rat liver cells (rIGFBP-2). Levels of rIGFBP-2 and IGF-II decrease in rat serum after birth. Using a recently isolated cDNA clone for rIGFBP-2 as hybridization probe, we now compare the expression of rIGFBP-2 and IGF-II in fetal tissues and the effects of hypophysectomy and fasting on the abundance of these mRNAs in adult rat liver. rIGFBP-2 mRNA is expressed at high levels in term gestation liver and at lower levels in other tissues. The ratio of rIGFBP-2 to IGF-II mRNAs in stomach, kidney, and lung is similar to that seen in liver, whereas IGF-II mRNA is more abundant than rIGFBP-2 mRNA in muscle, intestine, heart, and skin. Both mRNAs are more abundant in fetal tissues than in the corresponding tissues from adult rats. Dexamethasone treatment of 4-day-old rats for 4 days caused a greater (90%) decrease in hepatic IGF-II mRNA than in rIGFBP-2 mRNA (50%), suggesting subtle differences in the developmental regulation of the two mRNAs. Even more striking differences were observed in the regulation of the two mRNAs in adult rats after hypophysectomy or fasting. Hepatic rIGFBP-2 mRNA was increased 10- to 20-fold compared to age-matched control rats, whereas IGF-II mRNA was not increased. A parallel increase in serum rIGFBP-2 was observed, suggesting that this regulation may result at least in part from the increased abundance of rIGFBP-2 mRNA. Thus, in addition to modulating the stimulation of growth and differentiation by IGF-II in fetal tissues, rIGFBP-2 may play a homeostatic role during catabolic states in the adult rat.     

肿瘤组织中 GPC3、Hepatocyte、CD34检测对 AFP 阴性肝细胞癌的辅助诊断作用

目的评价肿瘤组织中磷脂酰肌醇蛋白聚糖3（GPC3）、Hepatocyte、CD,。蛋白检测对甲胎蛋白（AFP）阴性肝细胞癌（HCC）的辅助诊断作用。方法回顾性分析48例癌细胞AFP阴性的HCC组织及其癌旁组织中GPC3、Hepatocyte、CD。蛋白表达特点。结果48例HCC及其癌旁组织中GPC3蛋白阳性率分别为91．7％（44／48）、2．1％（1／48）,两者比较P〈0．01;Hepatocyte蛋白阳性率分别为95．8％（46／48）、100．0％（48／48）,两者比较P〉0．05;CD3。蛋白阳性率分别为100．0％（48／48）、4．2％（2／48）,两者比较P〈0．01。HCC组织中GPC3与Hepmo—cyte表达呈正相关（r=0．314,P〈0．05）。结论检测肿瘤组织中GPC3、Hepatocyte和CD。蛋白有助于AFP阴性原发性HCC的诊断。     

Insulin-like growth factor-I and -II messenger RNA expression in muscle, heart, and liver of streptozotocin-diabetic swine

We have investigated the effects of streptozotocin-diabetes and fasting in juvenile swine by monitoring IGF-I and -II gene expression in muscle, heart, and liver tissues. In diabetic pigs, IGF-I messenger RNAs (mRNA) were decreased by 50% in muscle and liver tissues, and by 70% in heart. The imposition of fasting on diabetic animals tended to further decrease IGF-I mRNA levels, and fasting alone also decreased IGF-I mRNA abundance in the three tissues (P less than 0.05). Insulin therapy restored IGF-I mRNA levels to normal in muscle and livers but was less effective in hearts of diabetic pigs. Relative IGF-I mRNA expression in heart and muscle tissues was 2-fold and 4-fold higher, respectively, than in liver tissues under normal conditions in these animals. Serum IGF-I concentrations and tissue extractable immunoreactive IGF-I levels were also measured. Serum IGF-I was markedly decreased in the diabetic state, dropping to 70% below control levels (P less than 0.01). Extractable IGF-I in liver declined by 50% with diabetes (P less than 0.01), and by 30% in muscle with diabetes and fasting (P less than 0.05), but no significant changes in heart levels of IGF-I protein were detected. Expression levels of IGF-II mRNAs in the three tissues were unaffected by diabetes or fasting. These results are consistent with earlier observations in rat liver and further demonstrate that IGF-I expression in muscle and heart is altered by diabetes and fasting, whereas IGF-II mRNAs do not change.     

肝细胞癌癌组织CBX7和PERK表达及其对手术治疗后患者预后的影响*

目的 研究肝细胞癌癌组织色素框同源蛋白7(CBX7)和磷酸化胞外信号调节激酶(PERK)表达及其对术后预后的影响。方法 2016年1月～2017年1月在我院接受治疗的肝细胞癌患者68例,所有患者接受肝癌切除术,并随访3年。术后,取癌组织和癌旁肝组织,采用RT-PCR法检测癌组织和癌旁组织CBX7 mRNA和PERK mRNA水平,采用免疫组化法检测癌组织CBX7和PERK蛋白表达。结果 癌组织CBX7 mRNA水平为(0.51±0.10),显著低于癌旁组织【(4.11±0.91),P<0.05】,而癌组织PERK mRNA水平为(4.87±0.99),显著高于癌旁组织【(0.58±0.15,P<0.05】;38例肿瘤直径>3 cm患者癌组织CBX7蛋白呈高表达阳性率为33.3%,显著低于30例肿瘤直径≤3 cm组的66.7%,而PERK高表达阳性率为73.3%,显著高于肿瘤直径≤3 cm组的26.7%,28例AJCC Ⅲ/Ⅳ期患者癌组织CBX7蛋白呈高表达阳性率为8.3%,显著低于40例AJCC Ⅰ/Ⅱ期患者的91.7%,而PERK高表达阳性率为70.0%,显著高于对照组的30.0%(P<0.05);32例癌组织CBX7高表达患者1 a、2 a和3 a生存率分别为78.1%、65.6%和53.1%,显著高于36例呈低表达患者【分别为52.7%、38.9%和30.6%,P<0.05】,38例癌组织PERK高表达患者1 a、2 a和3 a生存率分别为55.3%、44.7%和31.6%,显著低于30例呈低表达患者【分别为80.0%、63.6%和53.3%,P<0.05】。结论 癌组织CBX7表达下调,而PERK表达增强的HCC患者预后差,需要给予特别的关注。     

Determination of High Mobility Group A1 (HMGA1) Expression in Hepatocellular Carcinoma: A Potential Prognostic Marker

Our objective was to investigate the expression of HMGA1 mRNA and protein in hepatocellular carcinoma (HCC) and the correlation between its expression and clinical pathological characteristics and prognosis. HMGA1 expression was determined at both the mRNA level and the protein level in 30 HCC tissues and their corresponding paracancer liver tissues (PCLTs) and 2 normal liver tissues by RT-PCR and IHC. Follow-up study was done on the 30 patients involved in this research. HMGA1 mRNA was detected in nine cases of HCC tissues and two PCLTs, for a positivity rate of 30% and 6.7%, respectively (P < 0.05), whereas no HMGA1 mRNA expression was found in normal liver tisssues. Clinicopathological analysis revealed that HMGA1 mRNA expression was significantly correlated with Edmondson's grade (P < 0.05). HMGA1 protein was detected in four HCC tissues by IHC and located mainly in the nuclei; no positive staining was found in PCLTs. Follow-up study showed that HMGA1 mRNA-positive patients had a higher risk of recurrence/metastasis and a shorter survival than negative cases (P < 0.05). Our findings indicate that HMGA1 may be involved in the carcinogenesis and invasiveness of HCC and the determination of HMGA1 can be of great value in predicting the prognosis of patients with HCC. This work was supported in part by National Key Technologies R&D Program Grant 2001BA703B04 and National Basic Research Program Grant 2004CB720303, Ministry of Science and Technology, People's Republic of China.     

CyclinD1、CyclinE、CDK6在原发性肝细胞癌中的表达

探讨细胞周期调控因子CyclinD1、CyclinE、CDK6在原发性肝细胞癌中的表达及意义。应用免疫组织化学、原位分子杂交和细胞图象分析技术检测原发性肝细胞癌组织及其对应的癌旁肝组织(各20例)、正常肝组织(5例)中CyclinD1、CyclinE和CDK6mRNA表达情况。结果显示:CyclinD1、CyclinE和CDK6mRNA在肝细胞癌组织中呈阳性表达,阳性率分别为70%、75%和25%,正常肝组织呈阴性表达;肝细胞癌组织中CyclinD1和CyclinE的阳性表达与癌旁肝组织、正常肝组织相比,差异均有显著性意义(P<0.05);CyclinD1和CyclinE的过表达在肝细胞癌的发生和发展中起着重要作用。