细胞外信号调节激酶在卵巢癌中的表达及临床意义

目的：研究细胞外信号调节激酶（ERK）在卵巢上皮癌中的表达及ERK激酶MEK1（丝裂原活化蛋白激酶激酶1）抑制剂PD98059对人卵巢癌细胞生长的抑制作用。方法：采用western blot技术和免疫组化方法．对50例上皮性卵巢癌组织中ERK．和ERK，蛋白的表达进行了研究，分别以不同浓度的PD98059作用于卵巢癌细胞不同时间．通过MTT比色法观察PD98059对细胞增殖的抑制作用：应用扫描、透射电镜、细胞DNA提取及电泳实验研究PD98059对卵巢癌细胞的诱导凋亡作用。结果：ERK1和ERK2在卵巢癌中有过度表达，与正常卵巢及卵巢良性肿瘤相比，其差异有显著性。Ⅲ～Ⅳ期表达高于Ⅰ～Ⅱ期．差异有显著性。PD98059使卵巢癌细胞增殖比明显下降．其下降程度随PD98059浓度增高和作用时间延长而增强：PD98059 100μM作用72h即可诱导卵巢癌细胞呈现典型的凋亡特征。结论：ERK的过度表达在卵巢癌的发生发展中起重要作用，PD98059具有抑制卵巢癌细胞生长和诱导凋亡作用     

极化调节相关蛋白在胆管癌中的表达及意义

Objective: To investigate the expressions of atypical protein kinase C 1 subtype(aPKC-1)and E-cadherin in cholangiocarcinoma, and analyze molecular mechanisms of the invasion and metastasis of cholangiocarcinoma.Methods: The expressions of aPKC-1 and E-cadherin in 9 specimens of benign bile duct tissues and 35 specimens of cholangiocarcinoma were detected by EnVision immunohistochemistry, and their correlations to the clinicopathologic characteristics and invasion of cholangiocarcinoma were analyzed.Results: The positive rate of aPKC-1 was significantly higher in cholangiocarcinoma than in benign bile duct tissues(68.6% vs.11.1%, P=0.006), while the positive rate of E-cadherin was significantly lower in cholangiocarcinoma than in benign bile duct tissues(37.1% vs.88.9%, P=0.016).aPKC-1 expression was negatively correlated to E-cadherin expression(r=-0.287, P＜0.05).aPKC-1 expression was positively and E-cadherin expression was negatively correlated to the differentiation and invasion of cholangiocarcinorna(P＜0.05).Conclusion: The expressions of aPKC-1 and E-cadherin may reflect the differentiation and invasive potential of cholangiocarcinoma.As a polar regulation-associated protein, aPKC-1 may play an important role in the invasion and metastasis of cholangiocarcinoma.     

细胞外信号调节激酶1/2在瘦素促进子宫内膜癌Ishikawa细胞增殖中的作用

背景与目的:流行病学研究提示瘦素与子宫内膜癌的发生有关,但其作用机制尚不清楚.瘦素作为促有丝分裂原,能够显著促进多种细胞的生长增殖.本研究的目的在于探讨细胞外信号调节激酶1/2(extraeellular signal-regulated kinase 1/2,ERK1/2)在瘦素促进子宫内膜癌细胞增殖中的作用.方法:免疫荧光染色检测Ishikawa细胞中瘦素受体的表达;于Ishikawa细胞中分别加入不同浓度的瘦素(0、10、50、100、150 ng/ml),作用不同时间(6、12、24h),MIT法检测各组细胞的增殖情况:同时应用ERK1/2激酶特异性抑制剂PD98059阻断ERK1/2磷酸化,观察其对瘦素促进Ishikawa细胞增殖的影响;应用免疫印迹技术检测100 ng/ml瘦素作用于Ishikawa细胞不同时间后(20、40、60 min)ERK1/2的活化水平(以P-ERK1/2与ERK1/2的比值表示).结果:免疫荧光检测结果证实Ishikawa细胞存在瘦素受体的表达:瘦素能明显促进Ishikawa细胞的增殖.在0～100 ng/ml范围内瘦素浓度越高.细胞增殖越显著,100 ng/ml瘦素作用24h其增殖效应最大(A值=0.73±0.02),100 ng/ml组与150 ng/ml组差异无统计学意义(P=0.129);PD98059能明显抑制瘦素对Ishikawa细胞的增殖作用.100 ng/ml瘦素和100μmol/L PD98059作用24h后,细胞增殖率为(6.88±0.86)%;Ishikawa细胞经100 ng/ml瘦素处理后,ERK1/2活化程度明显增高.结论:瘦素可能通过激活ERK1/2信号转导途径促进子宫内膜癌细胞的增殖.     

RAS/RAF/丝裂原活化蛋白激酶/细胞外信号调节激酶信号通路在胃癌中的研究进展

胃癌（GC）的发病机制复杂,与多种因素有关,包括环境和遗传因素等。细胞内信号通路失调代表了一种常见的致病机制,可能适合对患者进行靶向药物治疗。RAS/RAF/丝裂原活化蛋白激酶（MEK）/细胞外信号调节激酶（ERK）信号通路在多种恶性肿瘤的发生发展中起到至关重要的作用。但迄今为止,大多数针对此通路的药物研究是基础实验,临床试验有待开展。本文就RAS/RAF/MEK/ERK信号通路在GC中的研究进展进行综述,重点论述其与GC的关系及潜在的作用机制,以及RAS/RAF/MEK/ERK信号通路抑制剂在GC治疗中的应用前景,旨在为GC的早期诊断和治疗提供新思路。     

整合素α5β1和细胞外信号调节激酶信号传导通路在非小细胞肺癌中的作用及其相关性研究

研究整合素α5β1 及磷酸化ERK1/2 （p-ERK1/2） 在非小细胞肺癌（NSCLC）组织中的表达及整合素α5β1 和ERK1/2 信号通路在人肺癌A549 细胞生长和迁移中的作用。方法：采用免疫组织化学法和免疫印迹（Western blot）法检测41 例NSCLC 组织和15例正常肺组织中整合素α5β1 及p-ERK1/2 的表达并分析两者的相关性,体外培养肺癌A549 细胞,以Anti-α5β1 或ERK 激酶抑制剂PD98059 作为工具药物,采用四甲基偶氮唑盐比色 （MTT） 法和 Annexin-V FITC PI 双染色法检测A549 增殖和凋亡,采用Western blot 检测A549 中MMP-9 蛋白水平。采用逆转录-聚合酶链反应（RT-PCR）和Western blot 检测Anti-α5β1 处理后A549 中ERK1/ 2 蛋白和mRNA 表达水平的变化。结果：人NSCLC 组织中整合素α5β1 及p-ERK1/2 阳性表达率（分别为58.53%,65.52%）明显高于正常肺组织（分别为26.67%, 20.00%）（P<0.01）。整合素α5β1与p-ERK1/2 表达均与病理分级（分别为P<0.001,P=0.030）、淋巴结转移（分别为P=0.014,P<0.001）、临床分期（分别为P=0.027,P=0.038）相关。整合素α5β1 与p-ERK1/2表达具有相关性（r=0.375, P<0.05）。经Anti-α5β1 或PD98059 处理后,A549 细胞增殖效应和早期凋亡细胞百分率增加,MMP-9 的表达均明显减弱。经Anti-α5β1 处理后A549 细胞中ERK 的mRNA 和蛋白表达受到抑制,ERK1/2 的磷酸化水平显著减少。结论： 整合素α5β1 及p-ERK1/2 在人NSCLC 组织中高表达,Anti-α5β1 在下调了细胞内ERK 的mRNA 和蛋白磷酸化的同时,可以明显抑制A549 细胞的增殖和MMP-9 蛋白表达,表明整合素α5β1 可能介导ERK1/2 信号转导通路参与了非小细胞肺癌中的异常增殖和迁移的调控。

     

p27及p57在皮肤鳞状细胞癌和Bowen病中的表达

Objective： To investigate the expression and clinical significance of cyclin dependent kinase inhibitors p27 and p57 in cutaneous squamous cell carcinoma and Bowen＇s disease. Methods： The expression of p27 and p57 were evaluated in 10 normal skin tissues, 25 Bowen＇s disease （BD） and 30 cutaneous squamous cell carcinoma （SCC） with immunohistochemistrical staining. Results： The levels of p27 protein expression in SCC were dramatically lower than those in normal skin and BD （P 〈 0.057. The levels of p57 protein expression in SCC were dramatically lower than those in normal skin and BD （P 〈 0.057. There was a posilk, e correlation between p27 and p57 （r = 0.469, P 〈 0.057. Conclusion： The decrease of p27 and skp2 may be used for considering the biological behavior of human cutaneous squamous cell carcinoma and Bowen＇s disease.     

Selective reduction of extracellular signal-regulated protein kinase (ERK) phosphorylation in squamous cell carcinoma of the larynx

Mitogen-activated protein kinase (MAPK) cascades transmit and amplify signals involved in cell proliferation as well as in cell death. In this study, the potential derangement of MAPK pathways has been evaluated in human squamous cell carcinomas (SCC) of the larynx. The expression and activity of the MAPK p38, ERK1/2p44/p42 and JNK/SAPKp46/p54 have been investigated by immunoblot analysis of tissue homogenates in 27 samples of primary laryngeal cancer and in 27 paired non-neoplastic laryngeal mucosa. On the same tissues, the activation of MAPK JNK/SAPKp46/p54 was also analyzed by an ELISA assay. The results obtained showed that both total and phosphorylated levels of JNK/SAPKp46/p54 and p38 were not different between tumor and normal samples. Conversely, while total protein levels for both ERK1p44 and ERK2p42 were not statistically different between tumor and normal samples, the analysis of the level of the activated forms of ERK1/2 showed a statistically significant decreased phosphorylation of both isoforms in the tumor samples compared to the control tissues. The rate of reduction was similar for both isoforms. Immunohistochemical analysis of all the activated MAPK (p38, JNK/SAPKp46/p54 and ERK1/2p44/p42) in both laryngeal SCC and normal mucosa demonstrated no difference of cellular localization. Activated ERK1/2p44/p42 and activated p38 demonstrated a nucleo-cytoplasmic distribution whereas activated JNK/SAPKp46/p54 were localized into the cytoplasmic membrane. The decreased activity of ERK1/2p44/42 in laryngeal SCC might reflect alterations in tumor suppressing activity or might derive from the interplay among various transduction pathways.     

非小细胞肺癌微血管密度测定及临床意义

目的探讨微血管密度(MVD)与非小细胞肺癌(NSCLC)肿瘤大小、TNM分期、血行侵袭转移、淋巴结转移及术后生存期的关系.方法53例NSCLC标本(腺癌27例,鳞癌26例)的石蜡切片,采用FⅧ-RA做为血管内皮标记物行免疫组化染色(SP法).在高倍(200X)视野下记数每张切片中的三个微血管高密度区内的微血管数,取其平均值做为MVD.结果MVD高的NSCLC易发生血行侵袭转移(P<0.05),术后生存期短(P<0.01);MVD高的鳞癌同时还易发生淋巴结转移(P<0.05).结论MVD高的NSCLC患者术易复发转移,生存期短.MVD是一个独立于TNM分期的重要的预后因子,对预测NSCLC术后早期的转移模式和指导术后的进一步治疗都具有重要的意义.     

磷脂酰肌醇-3激酶与直肠癌发生发展的相关性研究

目的:探讨磷脂酰肌醇-3激酶(Phosphoinositide-3kinase PI3K)与直肠癌发生及发展的关系,试图进一步探讨直肠癌发生及其转移机制,并为直肠癌的治疗开辟新的途径。方法:流式细胞仪检测PI3K的蛋白表达和直肠癌细胞增殖指数和免疫组化检测PI3K在正常和直肠癌组织中的表达与分布。结果:60例直肠癌组织中有48例增殖指数高于正常组织,差异有统计学意义。60例癌组织中PI3K的蛋白表达均高于正常直肠组织,差异亦有统计学意义。结论:在直肠癌组织中PI3K的蛋白表达和细胞增殖活性均高于正常组织,并且,随着肿瘤恶性程度的增加而逐渐升高,提示PI3K在直肠癌的发生和转移中具有一定的作用。     

Critical role of extracellular signal-regulated kinase (ERK) phosphorylation in novel vitamin K analog-induced cell death.

In the present study, we show that 2-(2-hydroxyethylsulfaryl)-3-methyl-1,4-naphthoquinone, or CPD 5, is a potent growth inhibitor for pancreas cancer cell lines (ID(50): 21.4 +/- 3.8, 31.8 +/- 2.7 and 55.2 +/- 4.5 microM for MiaPaCa, Panc-1 and BxPc3, respectively). It induced protein tyrosine phosphor-ylation of hepatocyte growth factor (HGF) receptor (c-Met) or epidermal growth factor receptor (EGFR), which increased progressively to a maximum level at 30 min in Panc-1 cells. The receptor phosphorylation by CPD 5 was indicated to be functional, since these receptors were found to bind with Grb2 or SOS1 protein. CPD 5 was also suggested to induce phosphorylation of external signal-regulated kinase (ERK). EGF induced cell proliferation through ERK phosphorylation, since U0126, which is an inhibitor of ERK phosphorylation, abrogated the increase of cyclin D1 by EGF. HGF increased the amount of p27 protein, suggesting that it is associated with cell differentiation. By contrast, U0126 reduced CPD 5-induced cell death. On two-dimensional electrophoresis, we found an extra type of phospho-ERK, and this was completely and selectively abolished by U0126. These results suggest that ERK phosphorylation, especially the extra spot on two-dimensional gel, is critically associated with CPD 5-mediated cell death.     

Inhibition of extracellular signal-regulated kinase (ERK) mediates cell cycle phase independent apoptosis in vinblastine-treated ML-1 cells

Chemotherapeutic agents induce alterations in intracellular signal transduction cascades that culminate in the initiation of the apoptotic program. Here, the relationship between the mitogen-activated protein kinase (MAPK) response and apoptosis in ML-1 cells treated with vinblastine and paclitaxel was investigated. We show that these compounds elicit different effects on MAPKs with vinblastine, but not paclitaxel, increasing both c-Jun-NH2-terminal kinase (JNK) and p38 activity. However, vinblastine and paclitaxel both induced apoptosis with similar kinetics, suggesting that increased JNK and p38 activity is not required for apoptosis that is induced by microtubule interfering agents. Strikingly, the abrogation of extracellular signal-regulated kinase (ERK)-signaling by the MAPK/ERK kinase (MEK)1/2 inhibitor PD098059 in combination with vinblastine robustly induced apoptosis in ML-1 cells at a rate much faster than treatment with vinblastine alone and occurred at all phases of the cell cycle. This apoptotic induction was attributed to JNK activation because: (a) non-JNK-activating concentrations of vinblastine failed to increase apoptosis in the presence of PD098059; (b) apoptosis induced by paclitaxel, which did not activate JNK, was not potentiated by PD098059; and (c) transduction of an inhibitor of JNK activity partially suppressed both JNK activity and apoptosis induced by vinblastine plus PD098059. Additionally, we found that the activation of JNK by vinblastine occurred upstream of effector caspase activation because treatment with a pan-specific caspase inhibitor (valine-alanine-aspartate-fluoromethylketone) resulted in complete abrogation of apoptosis with no effect on MAPK signaling. Taken together, these data suggest that inhibition of the MEK-->ERK signal transduction cascade alleviates cell cycle dependence for vinblastine-induced apoptosis by a mechanism that requires JNK activation.     

青年人直肠癌的临床病理特征

目的:探讨青年直肠癌患者的临床病理特征。方法:回顾性分析2002年1月-2012年1月10年收治、行手术治疗的279例直肠癌患者的临床病理特征并进行回顾性分析,以40岁为界限,分为青年组(年龄≤40岁)和非青年组(年龄>40岁)。结果:肿瘤下缘距肛门距离≤7cm的比例占73.5%(205/279),病理类型以腺癌为主,占79.6%(222/279),细胞分化多呈高分化(33.3%,93/279)和中分化(43.0%,120/279),肝转移比率为10.0%(28/279),手术方式以根治术为主,占85.7%(239/279)。青年组患者直肠癌比率为15.1%(42/279),青年组患者黏液腺癌+印戒细胞癌比例、细胞低分化比例、Dukes D分期比例、淋巴结转移比例、肝转移比例及姑息切除手术和单纯结肠造瘘,均显著高于非青年组(P<0.01)。青年组患者肿瘤下缘距肛门距离≤7cm的比例为81.0%(34/42),高于非青年组,差异不显著(P>0.05)。结论:青年直肠癌患者具有直肠癌组织低分化、恶性程度高、易发生淋巴结转移和肝转移、就诊时分期偏晚、根治手术比例较低等临床病理特征。     

Shp2 SUMOylation promotes ERK activation and hepatocellular carcinoma development

Shp2, an ubiquitously expressed protein tyrosine phosphatase, is essential for regulation of Ras/ERK signaling pathway and tumorigenesis. Here we report that Shp2 is modified by SUMO1 at lysine residue 590 (K⁵⁹⁰) in its C-terminus, which is reduced by SUMO1-specific protease SENP1. Analysis of wild-type Shp2 and SUMOylation-defective Shp2^K590R mutant reveals that SUMOylation of Shp2 promotes EGF-stimulated ERK signaling pathway and increases anchorage-independent cell growth and xenografted tumor growth of hepatocellular carcinoma (HCC) cell lines. Furthermore, we find that mutant Shp2^K590R reduces its binding with the scaffolding protein Gab1, and consistent with this, knockdown of SENP1 increased the interaction between Shp2 and Gab1. More surprisingly, we show that human Shp2 (hShp2) and mouse Shp2 (mShp2) have differential effects on ERK activation as a result of different SUMOylation level, which is due to the event of K⁵⁹⁰ at hShp2 substituted by R⁵⁹⁴ at mShp2. In summary, our data demonstrate that SUMOylation of Shp2 promotes ERK activation via facilitating the formation of Shp2-Gab1 complex and thereby accelerates HCC cell and tumor growth, which presents a novel regulatory mechanism underlying Shp2 in regulation of HCC development.     

食管鳞癌微血管密度检测的临床意义

目的　评价食管鳞癌组织内微血管密度与肿瘤进展和预后之间的关系。方法　采用免疫组化法检测 72例手术切除的食管鳞癌组织的微血管密度并对其与常见临床病理因素和预后之间的关系进行回顾性分析。结果　全组病例微血管密度值为 2 3 .74± 9.49/ 2 0 0倍视野 ,N1组、姑息性切除组、术后复发转移组和Ⅲ期 +Ⅳ期患者的微血管密度分别高于N0 组、根治性切除组、无复发转移组和Ⅱ期患者 (P <0 .0 5 ) ,高微血管密度组的术后生存曲线较低微血管密度组恶劣 (Logranktest ,P <0 .0 5 )。多因素分析表明微血管密度是影响预后的独立因素。 结论　血管形成可促进食管鳞癌的进展 ,并导致术后复发转移率升高及生存率下降 ,检测微血管密度具有判断术后复发转移的风险及预后的价值。     

血管生成素-2/受体-2在乳腺癌中的表达及在血管新生中的可能作用

目的：检测乳腺癌组织中血管生成素2（Ang-2）及其受体Tie-2的表达水平,探讨二者在乳腺癌发生发展及血管生成中的调节作用。方法：采用免疫组化（SP法）对79例乳腺癌标本、68例癌旁组织及42例乳腺纤维腺瘤组织中Ang-2、Tie-2及CD34标记的微血管密度（MVD）进行检测,分析它们与乳腺癌血管生成的关系。结果：乳腺癌组织中Ang-2、Tie-2阳性率显著高于乳腺纤维腺瘤和癌旁正常组织中的阳性率（P〈0.05）,在Ⅲ期乳腺癌中的阳性表达率高于Ⅰ-Ⅱ期（P〈0.05）,在有腋窝淋巴结转移组中的阳性表达率高于无淋巴结转移组（P〈0.05）;Ang-2、Tie-2的表达与MVD均呈正相关（P〈0.05）。结论：Ang-2和Tie-2在乳腺癌血管生成和进展中可能起重要作用。