P-p38和uPA蛋白在大肠癌中的表达及临床意义

目的：研究P-p38和uPA蛋白的表达与大肠癌生物学行为及预后的关系。方法：应用免疫组织化学（ SP）法检测P-p38和uPA在62例大肠癌、25例大肠腺瘤及18例正常大肠黏膜组织中的表达。结果：大肠癌组织中P-p38和uPA阳性表达率分别为79.0%和74.2%,均明显高于大肠腺瘤及正常大肠黏膜组织,差异有显著统计学意义（ P<0.05）。P-p38和uPA表达与大肠癌的分化程度、淋巴结转移、Dukeˊs分期及浸润肌层的深度密切相关（ P<0.05）,且两者表达成明显正相关（ P<0.05）。P-p38和uPA表达阳性者术后5年生存率明显低于阴性者,差异有显著意义（ P<0.05）。结论：P-p38和uPA 的高表达预示着大肠癌的低分化,并共同促进了肿瘤的侵袭、转移。联合检测P-p38和uPA 的表达情况对于评估大肠癌的发展及预后具有重要意义。     

乙型肝炎病毒X蛋白对肝癌细胞MKK3,p38MAPK表达的影响

目的：观察乙型肝炎病毒X蛋白对肝癌细胞中丝裂原活化蛋白激酶（MKK3）及其下游分子p38丝裂酶原活化的蛋白激酶（p38MAPK）的表达的影响,探讨相关的信号转导机制,及其对肝癌细胞生物学行为的影响。方法：分别将质粒pCDNA3.1及pCDNA3.1-HBx重组质粒经脂质体介导转染人肝癌细胞株HepG2,G418筛选培养,并用反转录PCR和Western blot鉴定,获得表达X蛋白的稳定细胞株HepG2-HBx和阴性对照细胞株HepG2-pCDNA3.1,以HepG2细胞株作空白对照。通过RT-PCR和Western blot检测上述三种细胞株中MKK3,p38MAPK在mRNA水平和蛋白水平的表达变化,并用细胞免疫荧光法检查磷酸化p38MAPK蛋白在细胞浆及细胞核中的变化;通过MTT实验检测三种细胞株的增殖情况。采用SPSS 12软件进行统计学分析。结果：MKK3在mRNA和总蛋白水平的表达在HepG2-HBx中均高于其他两组,其他两组无明显差异;p38MAPK在mRNA和总蛋白水平三组无明显差异,而其蛋白磷酸化水平和在核蛋白中的表达在HepG2-HBx中较其他两组升高;HepG2-HBx较其他两组细胞有更强的增殖能力。结论：HBx可以通过上调肝癌细胞中MKK3的表达,促进p38MAPK磷酸化和入核,从而进一步激活下游分子发挥生物活性。p38MAPK通路在HBx促进肝癌细胞的增殖中发挥重要作用。可能是导致HBV相关性肝癌与非HBV相关性肝癌临床特点及肿瘤生物学差异的机制之一。     

乙型肝炎病毒X蛋白对肝癌细胞MKK3,p38MAPK表达的影响

目的:观察乙型肝炎病毒X蛋白对肝癌细胞中丝裂原活化蛋白激酶(MKK3)及其下游分子p38丝裂酶原活化的蛋白激酶(p38MAPK)的表达的影响,探讨相关的信号转导机制,及其对肝癌细胞生物学行为的影响。方法:分别将质粒pCDNA3.1及pCDNA3.1-HBx重组质粒经脂质体介导转染人肝癌细胞株HepG2,G418筛选培养,并用反转录PCR和Western blot鉴定,获得表达X蛋白的稳定细胞株HepG2-HBx和阴性对照细胞株HepG2-pCDNA3.1,以HepG2细胞株作空白对照。通过RT-PCR和Western blot检测上述三种细胞株中MKK3,p38MAPK在mRNA水平和蛋白水平的表达变化,并用细胞免疫荧光法检查磷酸化p38MAPK蛋白在细胞浆及细胞核中的变化;通过MTT实验检测三种细胞株的增殖情况。采用SPSS 12软件进行统计学分析。结果:MKK3在mRNA和总蛋白水平的表达在HepG2-HBx中均高于其他两组,其他两组无明显差异;p38MAPK在mRNA和总蛋白水平三组无明显差异,而其蛋白磷酸化水平和在核蛋白中的表达在HepG2-HBx中较其他两组升高;HepG2-HBx较其他两组细胞有更强的增殖能力。结论:HBx可以通过上调肝癌细胞中MKK3的表达,促进p38MAPK磷酸化和入核,从而进一步激活下游分子发挥生物活性。p38MAPK通路在HBx促进肝癌细胞的增殖中发挥重要作用。可能是导致HBV相关性肝癌与非HBV相关性肝癌临床特点及肿瘤生物学差异的机制之一。     

P38MAPK信号通路与uPA在卵巢癌细胞及组织中表达的相关性

背景与目的：P38丝裂原活化蛋白激酶(P38 mitogen-activated protein kinase,P38MAPK)信号通路参与多种肿瘤的发生、发展和转移过程,尿激酶型纤溶酶原激活剂(urokinase-type plasminogen activator,uPA)在肿瘤浸润和转移中发挥着重要作用。本实验研究卵巢癌组织中P38MAPK、细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)、丝氨酸苏氨酸蛋白激酶(serine threonine kinase,AKT)及uPA的表达与临床病理特征的关系,并分析上述蛋白与uPA表达的相关性,探讨P38MAPK信号通路与uPA在卵巢癌细胞及组织中的表达及临床意义。方法：应用免疫组织化学法检测49例卵巢癌组织中uPA、P38MAPK、ERK和AKT蛋白的表达,采用蛋白［质］印迹法(Western blot)检测不同卵巢癌细胞系HO8910、HO-8910PM、SKOV3和CAOV3中uPA和P38MAPK蛋白的表达,使用特异性抑制剂SB203580阻断P38MAPK信号通路后检测uPA蛋白表达水平的变化。结果：uPA、P38MAPK、ERK和AKT蛋白在卵巢癌组织中的表达阳性率分别为61.22%、57.14%、53.06%和55.10%。uPA蛋白的表达与P38MAPK呈正相关(r=0.865,P=0.001),且与卵巢癌组织的临床病理分期(P=0.029)、分化(P=0.03)和转移程度(P_淋巴=0.022,P_大网膜=0.012)有关,而与患者的年龄(P=0.754)及组织学类型(P=0.652)无关。ERK、AKT蛋白的表达与卵巢癌淋巴结转移(P_ERK=0.011,P_AKT=0.022)和大网膜转移(P_ERK=0.006,P_AKT=0.000)有关,而与患者的年龄(P_ERK=0.000,P_AKT=0.022)、组织类型(P_ERK=0.771,P_AKT=0.245)及病理分期(P_ERK=1.000,P_AKT=0.254)无关。卵巢癌细胞系HO-8910PM中uPA蛋白的表达水平明显高于HO8910、SKOV3和CAOV3细胞系,使用SB203580阻断P38MAPK信号通路后可降低uPA蛋白的表达,且随着SB203580浓度升高uPA蛋白表达水平逐渐降低。卵巢癌中P38MAPK及uPA蛋白的表达与卵巢癌的预后显著相关(Log-rank=3.897和11.044,P=0.048和0.001)。结论：卵巢癌组织中P38MAPK信号通路处于激活状态;P38MAPK信号通路的激活可上调uPA的表达,促进卵巢癌的恶性进展;P38MAPK信号通路和uPA可能在卵巢癌侵袭和转移的过程中发挥重要作用。P38MAPK和uPA蛋白有望成为卵巢癌预后评估的重要指标。     

p38MAPK在肺鳞癌组织中的表达及临床意义

目的:探讨丝裂原活化蛋白激酶p38(p38MAPK)在肺鳞癌组织中的表达及意义。方法:采用real-time PCR检测肺鳞癌(lung squamous cell carcinoma,LSCC)患者癌组织和癌旁正常肺组织(non-tumor adjacenttissues,NAT)中p38MAPK mRNA表达水平;Western blot法检测p38MAPK(p38)和磷酸化p38MAPK(P-p38)蛋白表达水平,免疫组织化学法检测p38和P-p38在肺鳞癌组织中的定位。应用SPSS12.0软件分析与临床病理参数的关系。结果:肺鳞癌组织中p38MAPK mRNA和蛋白表达水平与对照组间均无明显差异(P>0.05)。癌旁正常肺组织中P-p38蛋白表达为(0.14±0.03),肺鳞癌组织中为(0.45±0.04),差异有统计学意义(P<0.05);P-p38蛋白表达与和肺鳞癌的组织分级程度呈正相关,与临床分期呈负相关(P<0.05)。结论:p38MAPK只有经过磷酸化活化后才发挥功能,即P-p38。P-p38可能在肺鳞癌的发生和发展中起到抑制作用,可以作为肺鳞癌的早期诊断和治疗的参考指标。     

p-p38和uPA在胃癌组织中的表达及意义

目的研究磷酸化p38（p—p38）和尿激酶型纤溶酶原潋活剂（uPA）在胃癌组织中的表达及与临床病理特征的关系。方法采用免疫组织化学方法检测35例胃癌组织标本中p—p38和uPA的表达并分析其相关性及与临床病理特征进行比较。结果胃癌组织中p-p38及uPA的阳性表达率分别为62,9％和60．0％。p—p38和uPA表达均为阳性17例,表达均为阴性9例,两者表达一致的符合率为74．3％（26／35）,其表达呈密切相关（P〈0．05）;两者与胃癌的浸润程度、淋巴结转移密切相关（P〈0．05）,而与患者年龄、性掣无显著相关性（P〉0-05）。结论p—p38及uPA过表达存胃癌的发展及浸润转移中具有重要的作用,p—p38和。PA的表达可辅     

子宫颈癌患者尿激酶型纤溶酶原激活物和抑制物的表达及其临床意义的研究

目的：研究纤溶酶原激活物和抑制物在子宫颈癌的发生、发展及侵袭转移过程中的规律及对临床的指导意义。方法：采用ELISA法检测42例子宫颈癌患者和10例良性子宫肿瘤患者的血浆和组织中uPA和PAI-1含量，按良恶性、手术前后、临床分期和组织类型等分别进行对照分析。结果：子宫颈癌患者血浆中uPA和PAI-1含量随临床分期的升高逐渐增加。宫颈癌患者血浆uPA和PAI-1含量术后显著降低。淋巴结转移组血浆uPA含量高于无转移组。癌组织中的uPA和PAI-1含量高于癌周组织。正常宫颈、宫颈上皮内瘤变、宫颈鳞癌组织uPA含量呈上升趋势。腺癌的组织uPA含量高于鳞状细胞癌组，PAI-1含量无差异。结论：检测宫颈癌患者uPA和PAI-1含量可能对其侵犯范围、盆腔淋巴结转移情况、预后等有一定的参考价值。     

尿激酶型纤溶酶原激活物系统在正常子宫内膜和子宫内膜癌中的表达

 尿激酶型纤溶酶原激活物系统包括尿激酶型纤溶原激活物 (Urokinase- type PlasminogenActivators,u PA)尿激酶型纤溶酶原激活物受体(Receptor for u PA,u PAR)、纤溶酶原激活物抑制剂 (Plasminogen Activator Inhibitor,PAI)。它们是体内生理或病理条件下通过降解胞外基质、介导细胞转移的主要酶系。其与肿瘤关系的研究近年来不断深入 ,本文将对尿激酶分子系统介导的肿瘤浸润与转移的机制 ,其在正常子宫内膜和子宫内膜癌中的表达等方面的进展作一综述。     

罗格列酮诱导人肝癌HepG2细胞凋亡中p38MAPK通路的作用北大核心CSCD

目的研究罗格列酮对人肝癌HepG2细胞增殖与凋亡的影响,探讨p38丝裂原活化蛋白激酶(p38MAPK)通路相关蛋白在其中发挥的作用。方法 MTT法检测罗格列酮对人肝癌HepG2细胞的增殖抑制率,流式细胞术检测细胞凋亡率,透射电子显微镜观察细胞凋亡形态变化,Western blot检测p38MAPK通路相关蛋白表达变化。结果罗格列酮可抑制HepG2细胞的增殖,诱导细胞凋亡(P<0.05);电子显微镜下可观察到典型的HepG2细胞凋亡表现。Western blot结果显示罗格列酮可激活p38MAPK通路,上调HepG2细胞中磷酸化p53及Bax蛋白的表达,下调Bcl-2蛋白的表达(P<0.05);而细胞外信号调节激酶ERK1/2的磷酸化程度没有明显改变。p38MAPK通路抑制剂SB203580可显著降低罗格列酮诱导的HepG2细胞的凋亡率;并且SB203580可部分逆转由罗格列酮引发的磷酸化p53、Bax及Bcl-2蛋白的表达变化。结论罗格列酮可通过激活p38MAPK通路诱导人肝癌HepG2细胞凋亡,其机制可能与p38MAPK通路参与磷酸化p53、Bax及Bcl-2蛋白的调控有关。     

t—PA,u—PA／uPAR及其抑制剂PAI—1在乳腺肿瘤的表达

目的　研究组织型纤溶酶原激活剂（ｔＰＡ） 、尿激酶型纤溶酶原激活剂（ｕＰＡ） 及其受体ｕＰＡＲ、１ 型纤溶酶原激活剂抑制剂ＰＡＩ１ 在乳腺肿瘤中的分布及酶学活性。　方法　用免疫组织化学ＡＢＣ方法,分析４ 种因子在乳腺肿瘤组织中的定位及半定量表达,用发色底物法测定乳腺肿瘤组织提取物中的ｔＰＡ 和ｕＰＡ 的活性。　结果　ｔＰＡ、ｕＰＡ、ＰＡＩ１ 主要分布在正常腺上皮及癌上皮细胞的胞质中,ｕＰＡＲ 则在癌细胞的胞膜及胞浆中;总ＰＡ 活性和ｔＰＡ 活性在乳腺良、恶性组织中均无显著性改变,但ｕＰＡ,ｕＰＡＲ 和ＰＡＩ１ 在乳腺癌组织中的表达明显高于良性组织及癌旁组织;ｕＰＡ／ｕＰＡＲ在转移灶中的表达较原发灶有不同程度的下降,而ＰＡＩ１ 在转移灶中却较原发灶中为高。　结论　ｔＰＡ 可能与乳腺癌的恶性表型无关。而ｕＰＡ／ｕＰＡＲ 与恶性表型相关;ｕＰＡ／ｕＰＡＲ 和ＰＡＩ１的相互制约及平衡在调节肿瘤原发灶癌细胞的移动中可能起到重要作用。     

CLINICAL STUDY OF T-PA AND U-PA EXPRESSION IN PATIENTS WITH GASTROINTESTINAL CANCER

Objective: To study the changes of tissue-type plasminogen activator (t-PA), urokinase-type plasminogen activator (u-PA) expressions and fibrinolysis molecular markers in patients with gastrointestinal cancer in order to elucidate their clinical significance. Methods: The plasma levels of t-PA, u-PA, urokinase-type plasminogen activator receptor (u-PAR) and plasmin anti-plasmin complex (PAP) were measured by ELISA. t-PA and u-PA mRNAs were detected by Real-time RT-PCR. Results: The plasma levels of u-PA, u-PAR and PAP were elevated in gastrointestinal cancer patients, while u-PA was markedly elevated in patients with local infiltration, lymph node involvement or distal metastasis. U-PA mRNA was higher and t-PA was lower in gastrointestinal cancer compared to normal tissue. Conclusion: Hyperfibrinolysis is an important factor related with metastasis potential of gastrointestinal cancer, t-PA may be a character of well differentiated tissue.     

促胃液素对结肠癌细胞侵袭力的影响

目的研究促胃液素对人结肠癌细胞侵袭力的影响,以及促胃液素-促胃液素受体-黏着斑激酶(FAK)信号通路在此过程中的作用。方法使用促胃液素受体的真核表达载体pCR3．1／GR转染人结肠癌细胞株Colo320(Colo320／GR),使其高表达促胃液素受体,达到上调信号通路的作用;使用反义寡核苷酸阻断FAK的表达,达到下调信号通路的作用。使用递增剂量的促胃液素(0,0．1,1,10,100nmol／L)刺激Colo320和Colo320／GR细胞,用Boyden小室检测各组细胞侵袭力变化;使用免疫沉淀和蛋白质印迹法,柃测细胞FAK第397位酪氨酸(tyr-397)磷酸化的状况。使用免疫沉淀、蛋白质印迹法和Boyden小室法检测正常细胞、Colo320／GR、转染反义寡核苷酸组细胞和对照组细胞在接受促胃液素刺激前后,细胞侵袭力和FAK tyr-397磷酸化的变化。结果促胃液素能够增强Colo320和Colo320／GR细胞侵袭力和FAK tyr-397磷酸化,具有剂量依赖性。促胃液素受体表达增加可以增强细胞的侵袭力和FAK tyr-397磷酸化。使用FAK反义寡核苷酸对FAK的表达进行阻断后,促胃液素的作用明显下降。结论促胃液素可有效地增强结肠癌细胞的侵袭力,其作用是通过促胃液素-促胃液素受体-FAK通路实现的。     

E-钙黏附素与尿激酶型纤溶酶激活物在浸润性乳腺癌中的表达及其临床意义

目的通过检测E-钙黏附素(ECD)和尿激酶型纤溶酶原激活物(UPA)基因产物在乳腺癌组织中的表达情况,探讨它们与乳腺癌浸润和转移及其与乳腺癌生物学行为的关系.方法采用免疫组化SABC方法检测ECD与 UPA在86例乳腺浸润癌(其中淋巴结转移62例,无淋巴结转移24例)中的表达,分析其与临床病理的关系及内在联系.结果淋巴结转移组UPA表达比无淋巴结转移组明显高(P<0.01),而ECD表达则减低(P<0.01).UPA的高表达与乳腺癌的高侵袭力、淋巴转移、组织低分化和病理分期升高有关,且ECD(-)/UPA( )组较ECD( )/UPA(-)组有明显的恶性生物学行为.结论 ECD表达减低和UPA高表达均提示其恶性生物学行为,有助于确定术后治疗方案.     

Inhibition of p38 MAPK increases the sensitivity of 5-fluorouracil-resistant SW480 human colon cancer cells to noscapine

A major cause of treatment failure in advanced colon cancer is resistance to chemotherapy. p38 mitogen-activated protein kinase (MAPK) has been associated with cellular apoptosis and plays an important role in multidrug resistance (MDR) in cancer cells. In the present study the effect of p38 MAPK on the sensitivity of 5-fluorouracil (5-FU)-resistant SW480 (SW480/5-FU) human colon cancer cells to noscapine was investigated. Following p38 MAPK interference, the inhibitory effect of noscapine on cell viability and proliferation was increased in the SW480/5-FU cells and there was also a decrease in the expression level of minichromosome maintenance proteins, recombinant Ki-67 and proliferating cell nuclear antigen. Inhibition of p38 MAPK also enhanced noscapine-induced G₁-phase cell cycle arrest in the SW480/5-FU cells and there was also a decrease in the protein and mRNA expression level of cyclin D, cyclin E and cyclin-dependent kinase 2, and an increase in the expression level of P57. Furthermore, p38 MAPK interference increased noscapine-induced apoptosis of the SW480/5-FU cells and there was an increase in the protein and mRNA expression level of caspases-3 and 8 and Bax, and decreased Bcl-2 expression level. The sensitivity of the SW480/5-FU cells to noscapine was also increased following p38 MAPK interference, as demonstrated by MDR inhibition via decreased Akt activity and reduced protein expression level of the MDR proteins P-glycoprotein, multidrug resistance protein 1 and ATP-binding cassette G2. These observations indicated that inhibition of p38 MAPK increased the sensitivity of the SW480/5-FU cells to noscapine by suppressing proliferation, induction of cell cycle arrest and apoptosis, and reversal of MDR in the SW480/5-FU cells.     

涎腺腺样囊性癌中丝裂原激活蛋白激酶p38的表达及意义

目的 探讨丝裂原激活蛋白激酶p38(p38MAPK)在涎腺腺样囊性癌(SACC)中的表达及临床意义.方法 采用组织微阵列平台,运用免疫组化和原位杂交技术,检测52例SACC和11例正常涎腺组织中p38MAPK蛋白和mRNA的表达,并分析其与SACC 临床病理特征的关系.结果 正常涎腺组织和SACC组织中,p38MAPK蛋白的阳性表达率分别为36.4%和96.2%,差异有统计学意义(P＜0.01).p38MAPK蛋白表达与SACC的淋巴结转移、神经侵犯有关(均P＜0.05).正常涎腺组织和SACC组织中,p38MAPK mRNA的阳性表达率分别为45.5%和94.2%,差异有统计学意义(P＜0.01).p38MAPK mRNA表达与SACC的淋巴结转移、神经侵犯有关(均P＜0.05).SACC组织中,p38MAPK蛋白与mRNA的表达呈正相关(r=0.409,P＜0.01).结论 在SACC组织中,p38MAPK表达上调,p38MAPK通路可能与SACC的发生、侵袭和转移有关.

Abstract：

Objective To investigate the expression of p38 mitogen-activated protein kinase (p38MAPK) in salivary adenoid cystic carcinoma (SACC) tissues and their clinicopathologic significance.Methods The protein and mRNA expressions of p38MAPK were examined by immunohistochemistry and in situ hybridization, respectively, in 52 cases of SACC and in 11 normal salivary gland tissues adjacent to the tumors on a tissue microarray platform.Results The positive rate of p38MAPK protein expression in the paracancerous normal salivary gland tissues and that in SACC were 36.4% and 96.2%, respectively,showing a significant statistical difference (P ＜ 0.01 ).The protein expression of p38MAPK was positively correlated with the lymph node metastasis and nerve involvement (P ＜ 0.05 ).The positive rates of p38MAPK mRNA in the paracancerous tissues and in the SACC tissues were 45.5% and 94.2%,respectively, with a significant statistical difference (P ＜0.01 ).The mRNA expression of p38MAPK was positively correlated with the lymph node metastasis and nerve involvement (P ＜ 0.05 ).In the SACC, there was a notable positive correlation between the p38MAPK protein and mRNA expressions (r = 0.409, P ＜0.01).Conclusions The expression of p38MAPK is up-regulated in salivary adenoid cystic carcinoma.p38MARK may be involved in the tumorigenesis, development and metastasis of this cancer.     

p38MAPK基因对人胶质瘤细胞BT325生长周期的影响

目的　研究 p3 8MAPK基因对人多形性胶质母细胞瘤BT3 2 5生长周期的影响。 方法　利用脂质体介导法将p3 8MAPK基因导入BT3 2 5细胞中 ,用免疫细胞化学染色和Western blot检测其在转染前后的表达情况 ,用HE染色、透射电镜和流式细胞仪等研究其对细胞形态和生长周期的影响。结果　pCMV 5 p3 8MAPK组p3 8MAPK蛋白表达阳性 ,细胞形态发生变化 ,G1%增多 ,而S %和G2 %减少 ,并出现凋亡峰。结论　p3 8MAPK基因可影响BT3 2 5细胞的生长周期 ,并诱导BT3 2 5细胞凋亡。     

A role for p38 MAPK in head and neck cancer cell growth and tumor-induced angiogenesis and lymphangiogenesis

We have recently gained a remarkable understanding of the mutational landscape of head and neck squamous cell carcinoma (HNSCC). However, the nature of the dysregulated signaling networks contributing to HNSCC progression is still poorly defined. Here, we have focused on the role of the family of mitogen activated kinases (MAPKs), extracellular regulated kinase (ERK), c-Jun terminal kinase (JNK) and p38 MAPK in HNSCC. Immunohistochemical analysis of a large collection of human HNSCC tissues revealed that the levels of the phosphorylated active form of ERK1/2 and JNK were elevated in less than 33% and 16% of the cases, respectively. Strikingly, however, high levels of active phospho-p38 were observed in most (79%) of hundreds of tissues analyzed. We explored the biological role of p38 in HNSCC cell lines using three independent approaches: treatment with a specific p38 inhibitor, SB203580; a retro-inhibition strategy consisting in the use of SB203580 combined with the expression of an inhibitor-insensitive mutant form of p38α; and short-hairpin RNAs (shRNAs) targeting p38α. We found that specific blockade of p38 signaling significantly inhibited the proliferation of HNSCC cells both in vitro and in vivo. Indeed, we observed that p38 inhibition in HNSCC cancer cells reduces cancer growth in tumor xenografts and a remarkable decrease in intratumoral blood and lymphatic vessels. We conclude that p38α functions as a positive regulator of HNSCC in the context of the tumor microenvironment, controlling cancer cell growth as well as tumor-induced angiogenesis and lymphangiogenesis.