HE4、NF-κBp65和MMP-9在卵巢上皮性肿瘤中的表达及其临床意义

目的探讨HE4、NF-κBp65以及MMP-9的表达与上皮性卵巢肿瘤临床病理生物学行为的关系以及对患者预后的影响。方法应用免疫组化对80例卵巢上皮性癌、10例交界性上皮性肿瘤及10例良性上皮性肿瘤组织进行HE4、NF-κB p65及MMP-9蛋白的检测。结果 HE4、NF-κBp65及MMP-9蛋白的阳性表达率在卵巢癌组均高于交界性及良性肿瘤组（P〈0.05）。三个指标的表达与EOC的组织学分级、腹腔脏器及淋巴结转移以及PTNM临床分期有关（P〈0.05）;在EOC中,NF-κBp65分别与HE4、MMP-9的表达呈正相关（r1=0.673,P〈0.05;r2=0.775,P〈0.05）。多因素分析,PTNM分期、MMP-9的表达是影响卵巢癌术后患者预后的独立因素（P〈0.05）;HE4阳性组与阴性组5年生存率分别为12.7%和84.0%,MMP-9阳性组与阴性组5年生存率分别为8.7%和70.6%,差异均有统计学意义（P〈0.05）。结论 NF-κBp65可能通过上调HE4、MMP-9的表达促进EOC的浸润和转移,联合检测HE4、NF-κBp65以及MMP-9的表达可能作为预测和评价患者预后的生物学指标。     

肝细胞癌中HIF-1α与PHD2的表达及临床意义

目的 研究HIF-1α和PHD2在肝细胞癌（HCC）组织中的表达,并分析其与临床病理特征间的相关性及临床意义.方法 应用免疫组化Envision法检测103例HCC组织、14例癌旁组织及13例正常肝组织中HIF-1α和PHD2的表达.结果在HCC、癌旁组织、正常组织中HIF-1α阳性表达率分别为63.1%、35.7%、0%,差异显著（P≤0.05）,其阳性表达与肿瘤大小、脉管瘤栓、分化程度、TNM分期均有显著差异 （P〈0.05）.PHD2在HCC组织中表达阳性率明显低于正常肝组织 （P≤0.05）.HCC中PHD2表达水平下调与肿瘤大小、分化程度、TNM分期的差异有统计学意义（P〈0.05）.生存分析显示HIF-1α阳性组与阴性组2年生存率分别为53.8%（35/65）、81.6%（31/38）,差异有统计学意义（P〈0.05）.Cox多因素分析显示肿瘤大小、脉管瘤栓、HBsAg阳性及HIF-1α的表达是影响HCC的独立预后因素.HIF-1α和PHD2表达呈正相关（r=0.219,P〈0.05）.结论 HIF-1α和PHD2的异常表达与HCC的一些恶性进展临床病理特征相关,提示其可能参与肝癌的发生、发展、侵袭,有助于肿瘤的预后分析.     

CD147 在前列腺癌中的表达及其与肿瘤临床病理特征的关系

目的:研究CD147在还没前列腺癌组织中的表达及其与肿瘤临床病理特征的关系。方法:选择2013年10月-2015年10月我院收治的前列腺癌患者61例作为研究对象,另选取同期接受手术治疗的前列腺增生患者49例作为对照组,术中收集前列腺癌患者的肿瘤组织和癌旁组织以及前列腺增生患者的组织标本,采用免疫组化法检测CD147在前列腺癌组织、癌旁组织及前列腺增生组织中的表达情况。结果:CD147在前列腺癌组织中的阳性表达率(95.08%)显著高于癌旁组织(32.79%)和前列腺增生组织(16.32%),差异具有统计学意义(P0.05);CD147在癌旁组织中的阳性表达率(32.79%)高于前列腺增生组织(16.32%),差异具有统计学意义(P0.05)。CD147的阳性表达与前列腺癌Gleason分级、临床分期、淋巴结转移及远处转移呈正相关关系(P0.05)。结论:CD147在前列腺癌组织中呈阳性表达,且Gleason病理分级≥5、临床分期T3~4、淋巴结转移N1及远处转移M1均为前列腺癌组织中CD147m RNA阳性表达的危险因素。     

P-gp、MMP-2、C-erbB-2在乳腺癌原发灶和转移淋巴结中的表达对比研究

目的：探讨乳腺癌侵袭转移和多药耐药之间的关系,为治疗方案的个体化提供依据。方法：采用免疫组化方法检测46例乳腺浸润性导管癌患者乳腺原发灶及相应腋淋巴结转移灶中P-gp、MMP-2、c-erbB-2的表达,结合临床表现、病理学指标,分析其相关性。结果：46例原发灶P-gp阳性表达35例（76.1%）,MMP-2阳性表达25例（54.3%）,c-erbB-2高表达18例（39.1%）;相应腋淋巴结转移灶P-gp阳性表达28例（60.9%）,MMP-2阳性表达16例（34.8%）,c-erbB-2高表达16例（34.8%）;P-gp、MMP-2蛋白表达水平与肿块大小、淋巴结转移数目均呈正相关（P〈0.05）,c-erbB-2蛋白表达水平与腋窝淋巴结转移数量呈正相关,与ER、PR表达呈负相关,P-gp阳性表达与MMP-2和c-erbB-2的表达呈正相关（P〈0.05）。结论：肿瘤原发灶与转移灶存在异质性,P-gp、MMP-2、c-erbB-2的表达与乳腺癌的多药耐药和侵袭转移有关,检测上述基因在原发灶与转移灶的表达,为乳腺癌选择个体化的化疗、内分泌治疗及分子靶向治疗提供了分子生物学依据。     

人脑胶质瘤中CD147 和MMP-2 的表达及临床意义

目的:通过探讨CD147和MMP-2在人脑胶质瘤中的表达及临床意义,为临床治疗提供参考。方法:选择2010年1月至2015年5月本院手术切除并经病理诊断确诊的脑胶质瘤标本70例作为观察组,根据WHO分型,将其分为低级别组31例和高级别组39例。另选取20例脑外伤患者并作内减压切除的标本作为对照组。采用免疫组化SP法检测CD147与MMP-2蛋白的表达,RT-PCR检测CD147mRNA表达,并探讨CD147表达与患者预后的关系。结果:观察组中CD147阳性表达率为75.71%,MMP-2阳性表达率74.29%,两者存在相关性(r=0.870,P0.05)。高级别组和低级别组CD147mRNA表达水平均强于对照组,差异有统计学意义(均P0.05),且高级别高于低级别组,差异有统计学意义(P0.05)。复发患者CD147阳性表达率显著高于非复发患者,差异有统计学意义(P0.05);生存时间5年患者CD147阳性表达率显著高于生存时间≥5年患者,差异有统计学意义(P0.05)。结论:人脑胶质瘤中CD147和MMP-2的表达与肿瘤恶性程度和患者预后密切相关,可为患者临床预后判断和临床治疗提供参考。     

下咽鳞状细胞癌组织claudin-1和MMP-9的表达及与临床病理特征及预后的关系研究

摘要 目的：研究下咽鳞状细胞癌组织紧密连接蛋白1（claudin-1）和基质金属蛋白酶-9（MMP-9）的表达及与临床病理特征及预后的关系。方法：选取2015年1月-2017年3月我院收集的75例下咽鳞状细胞癌组织标本进行研究,同期选取60例癌旁正常黏膜组织标本作为对照。采用免疫组织化学法检测claudin-1、MMP-9在下咽鳞状细胞癌组织与癌旁正常黏膜组织中的表达差异,分析claudin-1、MMP-9阳性表达与临床病理特征关系;Spearman相关性分析癌组织claudin-1与MMP-9的相关性。对患者进行5年随访,分析claudin-1、MMP-9表达与预后的关系。结果：下咽鳞状细胞癌组织claudin-1、MMP-9阳性表达率高于癌旁正常黏膜组织（P＜0.05）。claudin-1、MMP-9阳性表达与下咽鳞状细胞癌患者组织分化程度、淋巴结转移有关（P＜0.05）。经Spearman相关性分析显示,下咽鳞状细胞癌组织中claudin-1阳性表达与MMP-9阳性表达呈正相关（rs= 0.463,P＜0.05）。术后随访5年,2例失访,73例患者获得随访。Kaplan-Meier生存曲线显示,claudin-1阳性表达患者的5年生存率为35.71%,低于阴性表达患者的66.67%（P＜0.05）,MMP-9阳性表达患者的5年生存率为34.85%,低于阴性表达患者的57.14%（P＜0.05）。结论：下咽鳞状细胞癌组织中claudin-1、MMP-9阳性表达升高,其表达与组织分化程度、淋巴结转移和预后不良有关,两者呈正相关,可能发挥协同作用促进肿瘤发生发展。     

Snail基因及其蛋白、E-cadherin蛋白在胃癌中的表达及其临床意义

目的观察Snail mRNA及其蛋白、E-cadherin蛋白在胃癌组织中的表达及其与胃癌临床病理特征的关系,并探讨它们在胃癌发生、发展中的作用及其临床应用价值。方法收集96例手术切除胃癌标本,同时取80例癌旁组织作为对照。应用免疫组织化学S-P法检测胃癌组织、癌旁组织中snail蛋白、E-cadherin蛋白的表达;运用原位分子杂交技术检测胃癌组织、癌旁组织中Snail mRNA的表达。结果（1）Snail蛋白在胃癌组织阳性率（83.3%）显著高于癌旁组织（41.25%）（P〈0.05）;高、中分化组Snail蛋白阳性表达率显著低于低分化组（P〈0.05）;Snail蛋白的阳性表达率在乳头状腺癌、管状腺癌及低分化腺癌与黏液癌之间差异有显著性（P〈0.05）;Snail蛋白的表达与胃癌浸润深度、淋巴结转移及远处转移有关（P〈0.05）,与性别、年龄、肿瘤大小、肿瘤部位及临床分期无关（P〉0.05）;（2）胃癌组织中snail mR-NA的阳性率（76%）显著高于癌旁组织（30%）（P〈0.05）;高、中分化组Snail mRNA阳性表达率显著低于低分化组（P〈0.05）;Snail mRNA的阳性表达率在乳头状腺癌、管状腺癌及低分化腺癌与黏液癌之间差异有显著性（P〈0.05）;Snail mRNA的表达与浸润深度及淋巴结转移有关（P〈0.05）,与性别、年龄、肿瘤大小、肿瘤部位、临床分期及远处转移无关（P〉0.05）;（3）E-cadherin蛋白在胃癌组织阳性率（37.5%）显著低于癌旁组织（100%）（P〈0.05）;高、中分化组E-cadherin蛋白阳性率显著高于低分化组（P〈0.05）;E-cadherin蛋白阳性率在乳头状腺癌、管状腺癌及低分化腺癌与黏液癌之间差异有显著性（P〈0.05）;E-cadherin蛋白的表达与胃癌浸润深度、淋巴结转移、临床分期及远处转移有关（P〈0.05）,与性别、年龄、肿瘤大小、肿瘤部位均无关（P〉0.05）;（4）胃癌组织中snail mRNA和snail蛋白的表达呈正相关（r=0.594,P〈0.05）;Snail蛋白和E-cadherin蛋白的表达呈负相关（r=-0.234,P〈0.05）。结论（1）E-cadher-in蛋白低表达与Snail蛋白高表达可能是胃黏膜恶性转变以及胃癌发生浸润转移的重要生物学标志;联合检测E-cadherin蛋白与Snail蛋白对预测胃癌浸润转移有重要意义。（2）Snail蛋白可能在转录水平上调控E-cadherin蛋白的表达。     

MMP-7和Survivin在结直肠癌中的表达及其临床意义

目的探讨Wnt信号通路中基质金属蛋白酶7（matrix melluoproteinases7,MMP-7）和凋亡抑制基因Survivin在结直肠癌组织中表达及其与临床病理特征的关系。方法应用免疫组织化学染色（Elivision）方法检测100例结直肠癌组织和60例癌旁正常黏膜组织中MMP-7和Survivin蛋白的表达。结果MMP-7蛋白在100例结直肠癌组织和60例癌旁正常黏膜组织中的阳性表达率分别为77．00％（77／100）和13．33％（8／60）,两组问差异有统计学意义（P〈0．01）;Survivin蛋白在100例结直肠癌组织和60例癌旁正常黏膜组织中的阳性表达率分别为65．00％（65／lOO）和15．00％（9／60）,两组问差异有统计学意义（P〈0．01）。MMP-7与Survivin蛋白阳性表达均与肿瘤的淋巴结转移和Dukes分期有关（P〈0．05）,此外,MMP-7蛋白在结直肠癌中的阳性表达也与肿瘤的浸润深度有关（P〈0．05）。而MMP-7与Survivin蛋白的阳性表达无相关性（r=0．097,P〉O．05）。结论MMP-7和Survivin在结直肠癌中的高表达可能与结直肠癌的发生、发展、浸润和转移等相关,检测癌组织中MMP-7和Survivin的表达有助于为结直肠癌的病情进展及预后判断提供帮助。     

MMP-2、MMP-9及P53的表达在肝移植后肿瘤复发与转移预测中的价值研究

目的：探讨MMP-2、MMP-9及P53的表达在预测肝癌行肝移植后肿瘤复发与转移中的价值。方法：选择肝癌行全肝移植患者85例,应用免疫组化方法检测其切除的肝癌组织中MMP-2、MMP-9及P53的表达,并对患者进行移植术后随访,观察术后肝癌复发与转移情况。分析MMP-2、MMP-9及P53的表达与移植后肝癌复发与转移之间的关系和意义。结果：肝癌组织中MMP-2、MMP-9及P53阳性表达率均明显高于癌旁组织（P〈0.01）。MMP-2表达与肝癌肿瘤直径、TNM分期、病理分级及门静脉癌栓显著相关性（P〈0.05）,MMP-9表达与肝癌肿瘤直径、TNM分期、门静脉癌栓及肿瘤包膜显著相关性（P〈0.05）,P53表达与肝癌肿瘤直径、门静脉癌栓及肿瘤包膜显著相关性（P〈0.05）。MMP-2、MMP-9在未复发和转移组中的表达明显低于复发和转移组（P〈0.05）,而P53在两组间的表达差异无显著性（P〉0.05）。肝癌组织中MMP-2、MMP-9高阳性表达均是预测肝癌患者肝移植后肿瘤复发和转移的独立预见因子,而P53未显示其具有预测意义。结论：MMP-2、MMP-9及P53在肝癌组织中呈高表达,MMP-2、MMP-9的表达水平可以有效预测肝癌患者肝移植后肿瘤复发和转移。     

CD133在胃肠间质瘤的表达及其临床意义

目的 探讨胃肠间质瘤（gastrointestinal stromal tumor,GIST）中的CD133的表达及其与GIST临床病理特征的关系.方法 采用免疫组织化学法,检测122例胃肠间质瘤患者组织中的CD133、CD117、CD34、DOG-1、KI-67的表达情况.结果 CD133、CD117、CD34 、DOG-1、KI-67的阳性表达率分别为74.6%（91/122）、98.4%（120/122）、86.9%（106/122）、95.1%（116/122）、47.5%（58/122）.CD133的表达水平与胃肠间质瘤危险度分组高低、核分裂像数目、肿瘤部位有关（P〈0.05）,而与患者性别、年龄、肿瘤大小无明显相关性（P〉0.05）.CD133的表达水平与DOG-1的表达水平无明显相关性（P〉0.05）,而与CD117、CD34、KI-67的表达水平呈正相关.结论 CD133蛋白的表达可能与GIST的恶性行为与预后有关,与CD117、CD34和DOG-1联合检测对于判断GIST的病理性质可能具有重要价值.     

Lack of evidence for caveolin-1 and CD147 interaction before and after bleomycin-induced lung injury

Immunohistochemical and in vitro studies indicate that caveolin-1, which occurs abundantly in alveolar epithelial type I cells and microvascular endothelial cells of the lung, is selectively downregulated in the alveolar epithelium following exposure to bleomycin. Bleomycin is also known to enhance the expression levels of metalloproteinases and of the metalloproteinase inducer CD147/EMMPRIN in lung cells. Experimental in vitro data has showed that MMP-inducing activity of CD147 is under the control of caveolin-1. We studied the effects of bleomycin on the expression of caveolin-1, CD147 and metalloproteinases using an alveolar epithelial rat cell line R3/1 with properties of both alveolar type I and type II cells and explanted rat lung slices. In parallel, retrospective samples of bleomycin-induced fibrosis in rats and mice as well as samples of wild type and caveolin-1 knockout animals were included for immunohistochemical comparison with in vitro data. Here we report that treatment with bleomycin downregulates caveolin-1 and increases CD147 and MMP-2 and -9 expression/activity in R3/1 cells using RT-PCR, Western blot analysis, MMP-2 activity assay and immunocytochemistry. Immunofluorescence double labeling revealed that caveolin-1 and CD147 were not colocalized in vitro. The in vitro findings were confirmed through immunohistochemical studies of the proteins in paraffin embedded precision-cut rat lung slices and in fibrotic rat lung tissues. The caveolin-1-negative hyperplastic ATII cells exhibited enhanced immunoreactivity for CD147 and MMP-2. Caveolin-1-negative ATI cells of fibrotic samples were mostly CD147 negative. There were no differences in the pulmonary expression of CD147 between the normal and caveolin-1 deficient animals. The results demonstrate that bleomycin-induced lung injury is associated with an increase in CD147 expression and MMP activity, particularly in alveolar epithelial cells. In addition, our data exclude any functional interaction between CD147 and alveolar epithelial caveolin-1.     

CD147 depletion down-regulates matrix metalloproteinase-11, vascular endothelial growth factor-A expression and the lymphatic metastasis potential of murine hepatocarcinoma Hca-F cells

Extracellular matrix metalloproteinase inducer (EMMPRIN, CD147), which is a plasma membrane glycoprotein enriched on the surface of many malignant tumors promotes adhesion, invasion and metastasis of tumor cells. In addition, tumor-associated CD147 also induces vascular endothelial growth factors (VEGFs) expression. To investigate the possible role of CD147 in the mouse hepatocarcinoma cell line Hca-F with highly metastatic potential in the lymph nodes, we used an RNA interference (RNAi) approach to silence CD147 expression. The results showed that CD147 depletion in Hca-F cells resulted in the significantly decreased expression of matrix metalloproteinase-11 (MMP-11), VEGF-A at both mRNA and protein levels. The reduced CD147 expression also attenuated the invasive, adhesive, metastatic ability of Hca-F cells to lymph nodes both in vitro and in vivo. Our current findings reveal that the tumor biological marker CD147 functionally mediates MMP-11, VEGF-A expression and tumor lymphatic metastasis.     

棕榈酸促进肝癌细胞侵袭转移的分子机制

目的:探讨棕榈酸(Palmiticacid,PA)对人肝癌细胞系SMMC-7721侵袭转移能力的影响,并通过检测肝癌细胞系中CD147-MMPs信号通路在PA影响下的变化,初探PA影响肝癌细胞侵袭转移的分子机制。方法:PA(0、20、50、100μM)作用SMMC-7721细胞后(8、16、24h),MTT法检测细胞增殖,划痕及Transwell实验评价细胞迁移侵袭能力,Western-blot及real-time PCR检测CD147蛋白及其mRNA的水平,ELISA检测基质金属蛋白酶(MMP-2,MMP-9)的水平。结果:与对照组相比,PA作用SMMC-7721细胞后,细胞存活率无显著差异(P0.05);细胞迁移和侵袭能力显著增高(P0.05);CD147蛋白及其mRNA的表达显著增高(P0.05);培养上清中MMP-9的浓度显著增高(P0.05),MMP-2的水平则无变化。不同的梯度组之间相比较,细胞迁移和侵袭能力、CD147的表达水平(蛋白及其mRNA)以及培养上清中MMP-9的浓度均随PA作用时间和作用剂量的增大而产生更显著的增高。结论:PA通过活化CD147-MMPs信号通路促进SMMC-7721细胞的迁移侵袭。     

Stealth siRNA against CD147 inhibits hepatocellular carcinoma cell metastatic properties

CD147 plays a critical role in the invasive and metastatic activity of hepatocellular carcinoma (HCC) cells by stimulating the surrounding fibroblasts to secrete matrix metalloproteinases (MMPs). Tumor cells adhesion to extracellular matrix (ECM) proteins is the first step to the tumor metastasis. MMPs degrade the ECM to promote tumor metastasis. The aim of this research was to investigate the inhibitory effects of stealth small interfering RNA (siRNA) against CD147 on HCC cell line (SMMC-7721) metastatic properties including invasion, adhesion to ECM, gelatinase production, focal adhesion kinase (FAK) and vinculin expression. Flow cytometry (FCM) and western blot assays were employed to detect the transfection efficiency of the stealth siRNA against CD147. Invasion assays and gelatin zymography were also used to detect the effects of stealth siRNA against CD147 on SMMC-7721 cells’ invasion and gelatinase production. The effects of stealth siRNA against CD147 on FAK and vinculiln expression in SMMC-7721 cells were also detected by western blot. The results showed that stealth siRNA against CD147 inhibited SMMC-7721 invasion, adhesion to ECM proteins, MMP-2 production, and FAK and vinculin expression. These findings indicate that CD147 is required for tumor cell invasion and adhesion. Perturbation of CD147 expression may have potential therapeutic uses in the prevention of MMP-2-dependent tumor invasion.     

PD-L1和CD147在口腔鳞癌中的表达与临床意义

目的:探讨口腔鳞癌组织中免疫共刺激分子PD-L1与细胞外基质蛋白酶诱导因子CD147的表达、两者的相关性及临床意义。方法:应用免疫组化技术检测66例口腔鳞癌组织及36例正常口腔黏膜组织中PD-L1和CD147的表达,分析PD-L1、CD147表达的相关性及二者与口腔鳞癌临床病理参数的关系。结果:PD-L1在口腔鳞癌组织中表达阳性率为68.18%(45/66),正常口腔黏膜组织中表达阳性率仅为16.67%(6/36);CD147在口腔鳞癌组织中表达阳性率为74.24%(49/66),明显高于其在正常口腔黏膜组织中的表达13.88%(5/36)。PD-L1和CD147两者在口腔鳞癌组织中阳性表达率与口腔黏膜组织相比均明显升高(P0.01)。统计学分析显示,PD-L1和CD147在口腔鳞癌组织中的高表达与患者的性别年龄、吸烟史及肿瘤的体积等因素无明显相关,但与TNM分期及鳞癌的组织分化程度紧密相关。口腔鳞癌组织中PD-L1与CD147两者相关性分析r=0.342,P值小于0.01,说明二者的表达呈显著正相关。结论:口腔鳞癌组织中PD-L1与CD147均呈高表达,并且二者的过度表达可能与口腔鳞癌的发生、发展关系密切,合并检测二者可能为OSCC的诊疗及预后指明新的方向,为口腔鳞癌的靶向治疗提供新的靶点。     

Expression of CD147 on phorbol-12-myris-tate-13-acetate (PMA)-treated U937 cells differentiating into foam cells

Monocytes, macrophages, and foam cells expressing CD147 can stimulate the production of several matrix metalloproteinases (MMPs) associated with the development of atherosclerosis. We defined the CD147 expression profile and examined the correlation between foam cell development and MMP-2, -9 expressions. Foam cells were derived from U937-stimulated macrophages using various concentrations of oxidized low-density lipoprotein (ox-LDL). PMA-stimulated U937 cells had a 4- to 5-fold increase in CD147 mRNA compared to undifferentiated monocytes and membrane-associated (mCD147) on foam cells decreased in response to ox-LDL in a dose-dependent manner compared to untreated macrophages. In contrast, ox-LDL treatment increased the levels of soluble CD147 (sCD147) and MMP-2, -9 in a dose-dependent manner. Our data suggested that monocyte differentiation up-regulated CD147 expression and lipid enrichment of foam cells had no effect on CD147 mRNA expression. Lipid loading in macrophages reduced mCD147 expression while increasing the levels of MMP-2, -9 and sCD147 in supernatants.     

Construction of high-density display of CD147 ectodomain on VCSM13 phage via gpVIII: effects of temperature, IPTG, and helper phage infection-period

Production of VCSM13 phage displaying a high density of CD147 ectodomain (CD147Ex) was achieved when culturing conditions were modulated. A phagemid expressing CD147Ex was constructed and used to produce phage display CD147Ex gpVIII fusion protein in TG1 Escherichia coli. Displaying of CD147Ex via gpVIII was successfully increased when growing the transformed TG1 at 25 degrees C with IPTG-stimulation. In addition to temperature and IPTG-stimulation, the VCSM13 helper phage infection-period particularly affected the insertion of CD147Ex into phage progeny. By sandwich ELISA, incorporation of the CD147Ex into phage particle was confirmed. The correct size of the CD147Ex-gpVIII fusion protein at 28kDa was demonstrated by Western immunoblotting. Multivalent display of CD147Ex on phage particles will be valuable in discovering its ligand partner.     

HAb18G/CD147 enhances the secretion of matrix metalloproteinases (MMP) via cGMP/NO-sensitive capacitative calcium entry (CCE) and accordingly attenuates adhesion ability of fibroblasts

The present study examined the effect of hepatoma-associated antigen HAb18G (homologous to CD147) expression on the NO/cGMP-regulated Ca2+ mobilization to induce matrix metalloproteinases (MMP) production and attenuate adhesion ability of mouse fibroblast NIH/3T3 cells. HAb18G/CD147 cDNA was transfected into fibroblast 3T3 cells to obtain a cell line stably expressing HAb18G/CD147, t3T3, as demonstrated by immunofluorescence staining and flow cytometry assays. 8-Bromo-cGMP inhibited the thapsigargin-induced Ca2+ entry in 3T3 cells, whereas an inhibitor of protein kinase G, KT5823 (1 microM), led to an increase in Ca2+ entry. Expression of HAb18G/CD147 in t3T3 cells decreased the inhibitory response to cGMP. A similar effect on the Ca2+ entry was observed in 3T3 cells in response to an NO donor, (+/-)-S-nitroso-N-acetylpenicillamine (SNAP). The inhibitory effect of SNAP on the thapsigargin-induced Ca2+ entry was also reduced in HAb18G/CD147-expressing t3T3 cells, indicating a role for HAb18G/CD 147 in NO/cGMP-regulated Ca2+ entry. Results of gelatin zymography assays showed that addition of extracellular Ca2+ induced MMP (MMP-2, MMP-9) release and activation in a dose-dependent manner, and expression of HAb18G/CD147 enhanced the secretion of MMP-2 and MMP-9 in 3T3 cells. 8-Bromo-cGMP and SNAP reduced the production of MMP in 3T3 cells but not in t3T3 with HAb18G/CD147 expression. RT-PCR experiments substantiated that the expression of MMP-2 and MMP-9 mRNA in HAb18G/CD 147-expressing t3T3 cell was significantly greater than that in 3T3 cells. Experiments investigating adhesion potentials demonstrated that HAb18G/CD147-expressing t3T3 cells pretreated with Ca2+ attached to Matrigel-coated culture plates significantly less efficiently than 3T3 cells. The proportion of attached cells could be increased by treatment with 8-bromo-cGMP and SNAP in 3T3 cells, but not in t3T3. These results suggest that HAb18G/CD147 attenuates adhesion potentials in fibroblasts by enhancing the secretion of MMP through NO/cGMP-sensitive capacitative Ca2+ entry.     

细胞外基质金属蛋白酶诱导分子(CD147)在胰腺癌细胞及胰腺星状细胞中的表达(英文)

目的:探讨细胞外基质金属蛋白酶诱导分子(CD147)在胰腺癌细胞(Panc-1)及胰腺星状细胞(PSCs)的表达。方法:应用QRT-PCR,免疫细胞化学和免疫印迹分析方法检测Panc-1和PSCs细胞中EMMRPIN的表达,应用脱糖基化试剂N-glycosidase F及Endoglycosidase H鉴定CD147糖基化形式。结果:CD147在Panc-1和PSCs细胞质膜及细胞质中高表达,通过脱糖基化法首次鉴定出胰腺癌细胞及胰腺星状细胞中CD147不同的糖基化修饰。结论:CD147的糖基化修饰具有细胞特异性,可能与细胞恶性程度相关。