Effect of phosphorylation of MAPK and Stat3 and expression of c-fos and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44~(MAPK), p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44~(MAPK), p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44~(MAPK) and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between P42/44~(MAPK) and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Raf/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44~(MAPK), c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

肝复康对肝纤维化大鼠肝组织JAK2和STAT3表达的影响

目的观察中药肝复康对肝纤维化大鼠肝组织Janus激酶2（JAK2）及信号转导子和转录激活子3（STAT3）表达的影响,并对其在JAK2/STAT3信号通路上治疗肝纤维化的作用机制进行初步探讨。方法采用10%四氯化碳皮下注射制备肝纤维化模型,于造模第9周给予肝复康治疗12周。通过测定血清中ALT、AST活性及白蛋白和总蛋白的含量来反映肝脏功能,HE染色法观察肝组织病理学变化,RT-PCR观察JAK2和STAT3 mR-NA的表达。结果经肝复康治疗后,中剂量治疗组与模型组相比较,肝脏的组织学和血清学指标均明显改善,肝组织JAK2、STAT3 mRNA的表达显著减少（P〈0.01）。结论 （1）JAK2/STAT3信号通路在肝纤维化的形成过程中起重要作用;（2）肝复康对肝纤维化有疗效,其作用机制可能与降低肝组织JAK2和STAT3的表达,阻断JAK2/STAT3信号通路有关。     

Effect of HCV NS3 protein on P53 protein expression in hepatocarcinogenesis

ＮＴＲＯＤＵＣＴＩＯＮＨｅｐａｔｏｃｅｌｕｌａｒｃａｒｃｉｎｏｍａ（ＨＣＣ）ｉｓｏｎｅｏｆｔｈｅｍｏｓｔｃｏｍｍｏｎｈｕｍａｎｃａｎｃｅｒｓｉｎｔｈｅｗｏｒｌｄ．Ｒｅｃｅｎｔｌｙ,ｔｈｅＨＣＶｉｎｆｅｃｔｉｏｎｗａｓｆｏｕｎｄｔｏｂｅａｎｅｔｉｏｌｏ...     

MAPK信号通路在肝癌发生发展及治疗中的作用

丝裂原活化蛋白激酶(MAPK)信号通路的异常活化与肝癌的发生、发展、转移密切相关。介绍了MAPK通路蛋白在肝癌中的表达及其在肝癌增殖、分化、转移中的作用,阐述了MAPK信号通路在肝癌治疗及预后评价中的价值。认为MAPK信号通路在肝癌的发生发展及治疗中发挥非常重要的作用,是肝癌治疗及预后评价的潜在分子靶点。     

神经生长因子对人脐静脉内皮细胞在体外形成管腔样结构的影响

目的探讨神经生长因子(NGF)对人脐静脉内皮细胞(HUVECs)在体外形成管腔样结构(LLSs)的影响及其信号机制。方法将接种在基质胶上的HUVECs分为空白对照组、NGF组、NGF中和抗体组、对照IgG组、NGF中和抗体+NGF组、对照IgG+NGF组,以相差显微镜直接观察LLSs形态,并通过二脒基苯基吲哚/鬼笔环肽免疫荧光染色观察LLSs的细胞组成。为证实NGF促LLSs形成的信号通路,向培养液中分别预先加入丝裂原活化蛋白激酶3个主要信号通路的特异性抑制剂(PD98059、SB203580及SP600125),观察上述3个信号通路对NGF促LLSs形成的影响,并应用Western blot检测NGF及c-Jun N末端激酶(JNK)特异性抑制剂SP600125对HUVECs JNK表达及活性的影响。结果 NGF组LLSs的平均面积较空白对照组明显增加(P<0.01)。NGF中和抗体+NGF组LLSs的平均面积较对照IgG+NGF组明显减少(P<0.01);NGF中和抗体组LLSs的平均面积较对照IgG组亦明显减少(P<0.01)。NGF可激活HUVECs的JNK信号通路,SP600125可显著减少NGF诱导的LLSs形成(P<0.01)。PD98059及SB203580对NGF诱导的LLSs形成均无明显影响(P>0.05)。结论NGF能促进HUVECs在体外形成LLSs,可能与激活JNK信号通路有关。     

睡眠剥夺对大鼠颞下颌关节髁突组织p38信号通路相关蛋白表达的影响

目的 探讨睡眠剥夺对大鼠颞下颌关节（TMJ）的破坏作用及对p38信号通路相关蛋白表达的影响.方法 将40只Wistar雄性大鼠随机分为SD组和对照组.SD组采用改良多平台法建立睡眠剥夺模型.于制模4d后处死两组大鼠,HE染色观察TMJ髁突组织病理学变化,免疫组化SP法检测TMJ髁突组织p38信号通路相关蛋白MKK6、p38表达.结果 SD组TMJ髁突表面可见部分胶原纤维水肿、松解,出现炎症反应;SD组MKK6、p38阳性细胞率均高于对照组;MKK6、p38蛋白表达阳性率均高于对照组（P＜0.05）.结论 大鼠睡眠剥夺时TMJ出现病理性改变,p38信号通路可能参与了TMJ关节破坏的病理过程.     

食管癌组织中STAT3通路相关调控基因的表达及临床意义

目的探讨转录信号传导子与激活子-3(Stat3)及CyclinD1、p53、c-fos在食管癌发生发展过程中的作用及意义。方法采用免疫组化方法检测100份食管癌组织及60份正常食管黏膜组织中Stat3、CyclinD1、p53及c-fos蛋白的表达情况。结果 Stat3蛋白在正常食管黏膜、食管癌组织中的阳性表达率分别为6.7%、94.0%,P<0.05;食管癌组织CyclinD1、p53、c-fos蛋白均明显高于正常食管组织(P<0.05)。Stat3与CyclinD1、p53、c-fos表达呈正相关(P<0.05)。结论 Stat3过度表达在食管癌发生发展过程中起重要作用;CyclinD1、p53、c-fos蛋白在调控食管癌癌变过程中可能起协同作用。     

哺乳动物雷帕霉素靶蛋白信号通路及其下游蛋白在胃癌组织中的活化及临床意义

目的 研究哺乳动物雷帕霉素靶蛋白(mTOR)信号通路相关蛋白mTOR和真核起始因子4E结合蛋白1(4E-BP1)在胃癌组织中的活化情况及其临床意义.方法 利用免疫组织化学技术观察38例手术患者胃癌组织中mTOR和4E-BP1的活化情况,采用卡方检验及Kruskal-Wallis检验分析磷酸化mTOR及4E-BP1在癌区、癌旁和正常胃黏膜组织中表达的差异,以及各种临床病理参数间的表达差异.结果 磷酸化mTOR在胃癌组织中的阳性表达率高于匹配的癌旁及正常组织[71.1 ％(27/38)比50.0％(19/38)比44.7 ％(17/38);x2=11.031,P=0.026],其下游的4E-BP1在胃癌组织中的阳性表达率也高于匹配的癌旁及正常组织[68.4％ (26/38)比57.9％ (22/38)比28.9％(11/38);x2=13.943,P=0.007].磷酸化mTOR和4E-BP1与肿瘤Lauren' s分型、浸润分期、分化程度、淋巴结转移及患者年龄等无关,但胃癌组织中活化的4E-BP1在不同大小肿瘤间表达差异有统计学意义(H=3.86,P＜0.05).结论 mTOR信号通路在胃癌发生中存在过度激活,其下游蛋白4E-BP1磷酸化程度在不同大小肿瘤中存在差异.     

磷酸化p38丝裂原活化蛋白激酶在急性肝衰竭小鼠模型及HBV相关慢加急性肝衰竭患者肝组织中的表达及意义磷酸化p38丝裂原活化蛋白激酶在急性肝衰竭小鼠模型及HBV相关慢加急性肝衰竭患者肝组织中的表达及意义

目的：研究磷酸化后的p38丝裂原活化蛋白激酶（p－p38MAPK）在氨基半乳糖（D－GalN）或脂多糖（LPS）诱导的急性肝衰竭小鼠模型以及HBV相关慢加急性肝衰竭（ACLF）患者肝脏中的表达及其意义。方法采用D－GalN／LPS诱导C57BL／6小鼠构建急性肝衰竭模型,分别在给药0、0．5、1、2、4、6、8 h设立实验组,每组4只,处死小鼠取肝组织标本进行HE染色观察肝组织结构的病理变化,分别运用蛋白免疫印迹技术半定量检测及免疫组化染色定位检测肝组织中p－p38MAPK的表达;同时对照研究p－p38MAPK在HBV－ACLF、乙型肝炎肝硬化、慢性乙型肝炎（CHB）患者肝组织中的表达情况。组间比较采用独立样本t检验。结果蛋白免疫印迹法检测p－p38MAPK在肝衰竭小鼠肝组织匀浆中的表达随时间变化持续增高,给药6 h组半定量分析表达量显著高于正常对照组,差异具有统计学意义（t＝－2.727,P＝0．034）。免疫组化染色结果显示随存活时间的延长,小鼠肝组织炎症程度逐渐加重,炎症早期主要为窦细胞表达p－p38MAPK,随着肝组织损害加重,肝细胞表达p－p38MAPK渐多,坏死肝组织附近分布大量p－p38MAPK阳性肝细胞;正常人肝组织中p－p38MAPK的表达量很低,CHB患者肝组织内可见浸润淋巴细胞及肝细胞表达p－p38MAPK,并且随病程进展呈增多趋势,与临床观察病变程度逐渐加重相一致。结论 D－GalN／LPS诱导的小鼠急性肝衰竭模型中,p－p38MAPK表达随肝组织损害加重,表明其在肝衰竭病变过程中占重要地位;在HBV－ACLF患者致病过程中,p－p38MAPK信号通路可能发挥重要作用。     

Cardiac expression and subcellular localization of the p38 mitogen-activated protein kinase member, stress-activated protein kinase-3 (SAPK3)

Despite the interest in the roles that mitogen-activated protein kinases (MAPKs) play in the heart, the role of the different MAPK isoforms has been relatively poorly defined. A third isoform of p38 MAPK, known variously as stress-activated protein kinase-3 (SAPK3), p38- gamma or ERK6, has been previously shown to differ from p38- alpha/ beta both in its molecular weight and its lack of inhibition by the compound SB203580. We have generated monoclonal antibodies with specificity for SAPK3 demonstrated by immunoblot analysis, immunofluorescence studies, and cloning of SAPK3 from a rat heart cDNA expression library. By immunoblotting, we confirmed high expression of SAPK3 in fast, slow and mixed fibre types of murine skeletal muscle and observed significant expression restricted to heart, lung, thymus and testes. In addition to expression in normal heart (human, mouse, rat, dog and pig), we observed constant expression in diseased human heart, as well as control and hypertrophic cultured neonatal rat cardiac myocytes. Immunolocalization in cultured cardiac myocytes followed by confocal microscopy showed punctate, non-nuclear SAPK3 staining. In contrast, p38- alpha/ beta staining was non-punctate and distributed throughout the cytosol and nucleus. Whereas treatment with Leptomycin B to prevent nuclear export processes promoted higher levels of p38- alpha/ beta staining in cardiac myocyte nuclei, there was no apparent change in SAPK3 localization under these conditions. These differences between p38- alpha/ beta and SAPK3 probably reflect the specialized functions of SAPK3 and emphasize the need to evaluate SAPK3 upstream activators and downstream targets in the heart.     

STAT3蛋白在肝癌组织中的表达及临床意义

目的探讨STAT3和p-STAT3蛋白在肝癌组织中的表达及其临床意义。方法采用免疫组织化学的方法检测了90例肝癌组织及其相应的癌旁组织中STAT3和p-STAT3蛋白的表达,分析STAT3和p-STAT3蛋白表达与临床病理特征及患者预后的关系。特征参数之间用χ2检验;采用Kaplan-Meier法进行生存分析,用Log-rank检验进行曲线间比较;运用Cox回归进行单因素及多因素生存分析。结果肝癌组织中STAT3主要在细胞质中表达,而p-STAT3主要表达在细胞核内;STAT3和p-STAT3在肝癌组织中阳性表达率明显高于癌旁组织;STAT3和p-STAT3蛋白的表达与卫星灶（P=0.033,P〈0.01）、血管浸润（P=0.046,P〈0.01）及AJCC分期（P〈0.01,P〈0.01）有关,与患者的性别（P=0.280,P=0.403）、年龄（P=0.432,P=0.844）、肿瘤大小（P=0.762,P=0.161）、肿瘤数量（P=0.301,P=0.326）、肿瘤的分化程度（P=0.753,P=0.910）及甲胎蛋白（P=0.441,P=0.080）比较差异无统计学意义;肝癌组织中STAT3和p-STAT3蛋白高表达的患者5年生存率明显低于低表达患者。结论 STAT3和p-STAT3蛋白可能参与了肝癌的浸润转移,有望成为一种新的肝癌预后参考指标。     

Effect of Boschniakia rossica on expression of GST-P, p53 and p21(ras)proteins in early stage of chemical hepatocarcinogenesis and its anti-inflammatory activities in rats

AIM:To investigate the effect of Boschniakia rossica (BR) extract on expression of GST-P, p53 and p21(ras) proteins in early stage of chemical hepatocarcinogenesis in rats and its anti-inflammatory activities.METHODS:The expression of tumor marker-placental form glutathione S-transferase (GST-P), p53 and p21(ras) proteins were investigated by immunohisto-chemical techniques and ABC method. Anti-inflammatory activities of BR were studied by xylene and croton oil-induced mouse ear edema, carrageenin, histamine and hot scald-induced rat pow edema, adjuvant-induced rat arthritis and cotton pellet induced mouse granuloma formation methods.RESULTS:The 500mg/kg of BR-H2O extract frac-tionated from BR-Methanol extract had inhibitory effect on the formation of DEN-induced GST-P-positive foci in rat liver (GST-P staining was 78% positive in DEN+AAF group vs 20% positive in DEN+AAF+BR group, P<0.05) and the expression of mutant p53 and p21(ras) protein was lower than that of hepatic preneoplastic lesions (33% and 22% positive respectively in DEN+AAF group vs negative in DEN+AAF+BR group). Both CH(2)Cl(2) and H(2)O extracts from BR had anti-inflamatory effect in xylene and crotonoil induced mouse ear edema (inhibitory rates were 26%-29% and 35%-59%, respectively). BR H(2)O extract exhibited inhibitory effect in carrageenin, histamine and hot scald-induced hind paw edema and adjuvant-induced arthritis in rats and cotton pellet-induced granuloma formation in mice.CONCLUSION:BR extract exhibited inhibitory effect on formation of preneoplastic hepatic foci in early stage of rat chemical hepato-carcinogenesis.Both CH(2)Cl(2) and H(2)O extracts from BR exerted anti-inflammatory effect in rats and mice.     

Function of apoptosis and expression of the proteins Bcl-2, p53 and C-myc in the development of gastric cancer

INTRODUCTION In China ,the incidence and mortality of gastric cancer rank the second among all cancers. Recent development of cancer [1-20].The aim of this study was investigat the insight of apoptosis and bcl-2, p53 and C-myc protein expression in the development of gastric cancer .     

STAT3和CyclinD1在宫颈癌组织中的表达及意义

目的探讨信号转导和转录激活因子3（STAT3）和细胞周期蛋白D1（CyclinD1）表达与宫颈癌生物学行为的关系及意义。方法用免疫组化S-P法检测12例正常宫颈、24例宫颈上皮内瘤变（CIN）、60例宫颈癌中STAT3与CyclinD1的表达。结果宫颈癌组织STAT3、CyclinD1的阳性表达率均明显高于正常宫颈组织与CIN组织（P〈0．01,〈0．05）;STAT3和CyclinD1的表达呈正相关（r=0．327,P〈0．05）;STAT3、CyclinD1异常表达率随着病理分级和临床分期的增加而增加,两者的表达均与淋巴转移有关（P〈0．01,〈O．05）。结论STAT3可能通过激活其下游靶基因CyclinD1,引起宫颈癌细胞异常增殖和分化失控。     

TAT-p38（AF）融合肽的构建及其对紫外线诱导的p38 MAPK通路激活的抑制作用

目的构建TAT蛋白转导域介导的p38（AF）MAPK蛋白转导系统,研究该蛋白转导系统对紫外线刺激引起的p38通路激活作用的影响。方法将p38无活性突变体p38（AF）亚克隆至含有TAT蛋白转导域的pET-14b-TAT质粒,原核表达、纯化His-TAT-p38（AF）融合蛋白。激酶实验检测His-TAT-p38（AF）自身磷酸化作用。将该蛋白与NHI/3T3细胞孵育,Western blot法检测该蛋白的蛋白转导功能;紫外线照射后,检测内源性p38及其底物ATF2的磷酸化水平。结果酶切、测序表明质粒构建正确;目的蛋白被正确表达;His-TAT-p38（AF）融合蛋白无自身磷酸化作用;His-TAT-p38（AF）蛋白以浓度依赖性方式高效转导入细胞;His-TAT-p38（AF）的导入抑制紫外线诱导的内源性p38及其底物ATF2的磷酸化。结论TAT蛋白转运系统能高效转导外源蛋白进入真核细胞;p38无活性突变体TAT-p38（AF）抑制紫外线诱导的p38 MAPK通路的激活。     

胃癌及癌前病变中幽门螺杆菌感染与Ezrin蛋白表达的关系

目的探讨细胞骨架蛋白Ezrin在胃癌、癌前病变中的表达与Hp感染关系,及其在胃癌发生发展中可能的机制。方法采用免疫组化方法检测49例胃癌及癌前病变组织及16例正常胃黏膜标本中Ezrin蛋白的表达情况,并用快速尿素酶试验或Warthin．Starry胃螺旋染色法检测幽门螺杆菌（Hp）感染。结果胃癌及癌前病变组织Hp感染率显著高于正常组织（P〈0．01）;Ezrin蛋白表达在正常胃组织、癌前病变组织和胃癌组织中呈递减趋势,三者比较差异有统计学意义（P均〈0．05）;胃癌组织及癌前病变组织中Hp感染阳性组Ezrin蛋白的表达显著低于阴性组（P〈0．05）。结论在胃黏膜癌变过程中,Ezrin蛋白表达逐渐下调,Ezrin蛋白表达与Hp感染相关,提示Hp感染可通过影响Ezrin蛋白表达,在胃癌的发生发展过程中发挥一定作用。     

Effect of boschniakia rossica on expression of GST-P, p53 and p21ras proteinsin early-stage chemical hepatocarcinogenesis and its anti-inflammatoryactivities in rats

AIM To investigate the effect of boschniakia rossica (BR) extract on expression of GST-P, p53 and p21fasproteins in early-stage chemical hepatocarcinogenesis in rats and its anti-inflammatory actions.METHODS The expression of tumor marker, placental form glutathione S-transferase (GST-P), p53 and p21ras proteins were investigated by immunohistochemical techniques and ABC method. Anti-inflammatoryactivities of BR were observed by xylene and croton oil-induced mouse ear edema, carrageenin, histamineand hot scald-induced rat pow edema, adjuvant-induced rat arthritis and cotton pellet-induced mousegranuloma formation methods.RESULTS The 500 mg/kg of BR-H2O extract fractionated from BR-Methanol extract had inhibitory effecton the formation of DEN-induced GST-P-positive foci in rat liver and the expression of mutant p53 and p21fasprotein was lower than that of hepatic preneoplastic lesions. Both CH2Cl2 and H2O extract from BR haveinhibitory effect in xylene and croton oil-induced mouse ear edema. BR-H2O extract exhibited inhibitoryeffect in carrageenin, histamine and hot scald-induced hind paw edema and adjuvant-induced arthritis in ratsand cotton pellet-induced granuloma formation in mice.CONCLUSION BR extract exhibited inhibitory effect on formation of preneoplastic hepatic foci in earlystage of rat chemical hepatocarcinogenesis. Both CH2Cl2 and H2O extract from BR exerted anti-inflammatory effect in rats and mice.     

组织芯片检测肝肠钙粘连蛋白在肝癌组织中的表达及临床意义

目的研究肝肠钙粘连蛋白（LI—cadherin）在肝癌组织中的表达以及与肝癌的发生发展和临床病理特征之间的相关性。方法采用免疫组织化学sP法检测70例不同类型肝癌组织和5例正常肝组织标本中LI—cadherin的表达情况,并结合临床资料和相关病理参数进行统计分析。结果70例肝癌组织中LI—cadherin阳性表达39例,总阳性率为55．7％,5例正常肝组织中未见阳性表达;LI—cadherin的表达与年龄、性别、肿瘤分化程度以及有无远处转移无关（P〉0．05）,而与淋巴结转移及血管侵犯密切相关（P〈0．05）。结论LI—cadherin的表达与肝癌的发生、转移和浸润有关,LI—cadherin可能成为诊断肝癌发生、转移及临床分期的新生物学标志物,对于提高临床治疗效果具有指导意义。     

STAT-3在大肠癌和小肠癌中的表达及意义

目的检测大、小肠癌中信号转导与转录活化因子3（STAT-3）的表达情况并探讨其相关性。方法用免疫组化法分别检测36例小肠癌和20例癌旁正常小肠黏膜,60例结直肠癌和22例癌旁正常大肠黏膜中STAT-3的表达情况。用半定量方法对免疫组化染色评分,结合临床和病理数据进行分析。结果 STAT-3定位于细胞质和胞核,在大、小肠癌中表达较癌旁正常组织增高（P〈0.01）;STAT-3表达与肠癌组织分化程度、淋巴结转移和TNM分期相关;STAT-3在大、小肠癌中的表达呈正相关（r=0.849,P〈0.05）。结论 STAT-3在肠癌组织中过度表达并与其发生发展有关,可能参与了大、小肠癌恶化的调控。