染料木素抑制HER2阳性人乳腺癌与卵巢癌细胞增殖作用的研究

目的研究染料木素对人HER2高表达乳腺癌与卵巢癌细胞增殖的抑制作用,并探讨其作用机制。方法采用流式细胞分析技术、western blotting及细胞增殖与凋亡检测试剂盒等检测方法,对染料木素作用于HER2表达阴性的MCF-7细胞与通过稳定转染HER2而获得的MCF-7-1以及BT-474、SKOV-3等HER2阳性表达细胞的效果进行了剂量或时间反应关系的研究。结果浓度为0.1～10μg/ml的染料木素对HER2表达水平低的MCF-7细胞增殖的影响不大,而对HER2高表达的MCF-7-1、BT-474和SKOV-3细胞的增殖有显著的抑制作用(与DMSO对照组相比,P<0.01),且表现出剂量依赖性反应关系。用10μg/ml的染料木素作用12～48 h可以诱导BT-474细胞凋亡或坏死(与DMSO对照组相比,P<0.01),且表现出时间依赖性反应关系。结论染料木素能够明显抑制HER2阳性肿瘤细胞的增殖,这一作用可能是通过下调肿瘤细胞HER2信号转导而介导的。     

溶血磷脂酸在卵巢癌诊断中的价值分析

目的明确溶血磷脂酸（LPA）在卵巢癌诊断中的作用。方法通过对2006～2008年人院的450例卵巢肿瘤患者进行手术前测定CA125和LPA,其中的恶性肿瘤患者于术后4周复测CA125和LPA以进行对比研究。结果相比较于CAl25,LPA在卵巢癌的诊断和随访中具有更高的诊断价值,二者的统计结果具有显著性差异（P〈0．05）。结论LPA在卵巢癌的诊断和随访中具有重要的价值。     

磷脂酶A2和溶血磷脂酸在卵巢癌诊断中的价值

目的探讨分泌性磷脂酶A2(sPLA2)和溶血磷脂酸(LPA)对卵巢癌诊断、病程监测和预后预测的价值.方法用 3H标记大肠杆菌膜为底物的液闪方法和比色法,分别对初发或复发卵巢癌和卵巢良性肿瘤患者各30例进行血液中sPLA2活性和LPA水平检测,并与乳腺癌患者作比较,同时对手术前后卵巢癌患者进行动态检测,以评价两项指标在卵巢癌检测中的敏感性和特异性.结果初发卵巢癌组与卵巢良性肿瘤组相比,sPLA2活性(24.47±5.64 U/L与4.94 ±1.86 U/L)和LPA水平(6.35 ±2.32与1.92 ±0.75 mmol/L)差异均具有统计学意义 (均P＜0.01).复发卵巢癌组与卵巢良性肿瘤组相比,sPLA2活性(26.53 ±6.05 U/L与4.94 ±1.86 U/L)和LPA水平(7.03 ±2.77 mmol/L与1.92 ±0.75 mmol/L)差异也均具有统计学意义(均P＜0.01).初发卵巢癌组手术前后sPLA2活性和LPA水平与时间变化相关(分别r=0.88,r=0.81,均P＜0.05).sPLA2活性和LPA水平的敏感性分别为100%和95%,特异性分别为73.3%和83.3%.初发和复发卵巢癌与初发和复发乳腺癌相比, LPA水平差异有统计学意义(P＜0.01).结论 sPLA2活性和LPA水平可能对卵巢癌的诊断、治疗监测和预后评价有一定的价值.     

白血病和卵巢癌亲代及耐药细胞系细胞外调节蛋白激酶与端粒酶活性的变化

本研究观察白血病和卵巢癌亲代及耐药细胞系中端粒酶活性及细胞外调节蛋白激酶(extracelluar regulated protein kinases ERK)磷酸化蛋白表达水平的变化，探讨端粒酶与ERK在白血病及卵巢癌耐药中的作用。采用MTT法评价白血病和卵巢癌亲代和耐药细胞系对HRT或DDP的敏感性，用流式细胞术分析这两种细胞系间细胞周期分布的差别，用端粒酶重复扩增技术(TRAP)、生物发光分析法定量和定性检测端粒酶活性及用Western blot检测法检测磷酸化ERK1和ERK2蛋白的表达水平。结果表明：白血病和卵巢癌耐药细胞系处于G0／G1期的细胞比例增加，端粒酶活性和磷酸化ERK1/2蛋白表达水平耐药细胞系较亲代细胞系高。结论：耐药细胞系G0／G1期细胞比例增加可能是细胞产生耐药的一种标志。白血病和卵巢癌细胞系耐药的产生可能与端粒酶活性和磷酸化ERK1/2蛋白表达水平的增高有关。     

溶血磷脂酸与CA125在复发性卵巢癌中的价值分析

目的 分析血浆溶血磷脂酸（lysophosphatidic acid,LPA）及CA125在卵巢癌复发患者中的表达情况和监测价值.方法 选择2013年1月-12月广州市第一人民医院收治的32例上皮性卵巢癌复发患者,测定血浆LPA和CA125表达情况,检测同期28例上皮性卵巢癌治疗后完全缓解患者、30例初诊上皮性卵巢癌患者及30例正常女性的血浆LPA和CA125作为对照,分析LPA和CA125在卵巢癌复发患者中的敏感度和特异度.结果 上皮性卵巢癌复发组血浆LPA（9.52±4.64 mol/L）明显高于完全缓解患者组（2.02±1.94 mol/L）（t=7.96,P＜0.05）及正常人群组（1.91±2.21 mol/L）（t=8.15,P＜0.05）,与初诊患者组（9.19±2.47 mol/L）差异无统计学意义（t=0.35,P＞0.05）,上皮性卵巢癌复发组血浆CA125（75.42±10.66 U/ml）明显高于完全缓解患者组（11.42±9.67 U/ml） （t=28.830,P＜0.05）及正常人群组（10.45±6.42 U/ml）（t=24.220,P＜0.05）,与初诊患者组（63.29±132.32 U/ml）差异无统计学意义（69.29±7.32 U/ml）（t=2.622,P＞0.05）.采用LPA和CA125诊断上皮性卵巢癌复发的诊断价值有差异（χ^2=37.834,P=0.022）,其中LPA敏感度（93.8％）高于CA125（65.6％）,特异度无明显差异（χ^2=24.501,P=0.99）.结论 LPA水平对判断上皮性卵巢癌的复发有重要价值,可能成为监测上皮性卵巢癌复发的新方法.     

HER2基因RNA干扰质粒对SK-BR-3细胞增殖及凋亡的影响

目的探讨HER2基因高效RNA干扰质粒转染到SK-BR-3乳腺癌细胞后,对细胞的增殖及其凋亡的影响。方法采用脂质体转染法将针对HER2基因的高效RNA干扰质粒转染至乳腺癌细胞SK-BR-3中,通过细胞计数、MTT比色法、流式细胞术和Western blot检测PCNA增殖细胞核抗原,分析检测其对SK-BR-3细胞增殖、凋亡的影响。结果 MTT比色测定显示,HER2-shRNA2干扰质粒转染到SK-BR-3细胞后在48 h后现抑制,与空白对照组比抑制率分别为48 h 16.53%7、2h 39.03%9、6h 65.47%。流式细胞分析HER2-shRNA组细胞在96 h凋亡率达17.36%,明显高于阴性对照组的1.41%和空白对照组的1.25%（P〈0.05）。结论将RNA干扰质粒HER2-shRNA2转染至SK-BR-3细胞中,可特异性地抑制SK-BR-3细胞的增殖并诱导其凋亡,为乳腺癌的靶向治疗奠定了基础。     

溶血磷脂酸对小胶质细胞活化和吞噬功能的影响

目的:观察溶血磷脂酸(LPA)对小胶质细胞活化及其吞噬功能的影响。方法:小胶质细胞系BV2细胞复苏后传代培养,取对数生长期细胞分为对照组和LPA组,LPA干预30min后各组加入FITC标记的葡聚糖,在干预后1h、2h后收取细胞,用流式细胞仪测定各孔FITC-葡聚糖微粒吞噬率及荧光强度。结果:LPA干预1h时,吞噬率较对照组明显增加(P<0.05),2h时两组细胞吞噬率达到最高值。LPA干预后1h及2h时,吞噬荧光强度较对照组明显增加(P<0.01),而且吞噬率与荧光强度的乘积较对照组亦明显增加(P<0.01)。结论:LPA可提高BV2细胞活化及吞噬功能,提示LPA在小胶质细胞活化及吞噬过程中发挥了重要作用。     

溶血磷脂酸在缺血性脑血管病中的作用

溶血磷脂酸（lysophosphatidic acid,LPA）是一种结构简单、具有多种生物活性的磷脂介质,作为血清中诱导成纤维细胞增生和应力纤维形成的成分而被首次发现。随后,人们发现LPA可以诱导其他多种类型细胞发生增生性和形态学改变。近年来研究发现,LPA在体内信号传递过程中起着十分重要的作用,是一种多功能的“磷脂信使”。通过G蛋白耦连受体影响靶细胞的功能,具有广泛而多样的生物学功能,其中部分生物学功能与缺血性脑血管病及其危险因素密切相关,血浆中LPA升高被认为是缺血性脑血管病的又一危险因子。LPA对促进动脉粥样硬化（Atherosclerosis,AS）发生发展、促进血小板聚集和脑血管痉挛以及对星形细胞有重要作用。笔者就溶血磷脂酸在缺血性脑血管病中的作用作一综述。     

溶血磷脂酸及CA125在卵巢癌诊断中的价值对比分析

目的 对比CA 125与溶血磷脂酸(LPA)在卵巢癌诊断中的作用.方法 对2006-2008年入院的450例卵巢肿瘤患者进行手术前测定CA 125和LPA,其中良性卵巢肿瘤408例,恶性卵巢肿瘤42例.恶性肿瘤患者于术后4周复测CA125和LPA以进行对比研究.结果 卵巢恶性肿瘤组血浆LPA(5.25±1.57)和CA125(159.6±58.7)U/L均高于卵巢良性肿瘤组[分别为(1.74±0.71)μmol/L],(11.10±6.89)U/L,恶性肿瘤患者二者均有明显的升高,差异有统计学意义(t值分别为12.032、8.205,P均＜0.05).LPA诊断准确率高于CA125(82.4%与36.0%,x2=3.96,P＜0.05).恶性肿瘤外周后复查CA125和LPA阳性率分别为74%与100%(x2=2.76,P＞0.05).结论 与CA125相比LPA在卵巢癌的诊断和随访中具有重要的价值.     

卵巢癌患者血清中LPA、CA125的变化对于判断其疗效及预后的临床研究

肿瘤标志物的作用除了早期发现肿瘤从而有利于早期治疗外,还有由于监测治疗活动的效果以及对患者的未来进行预测的作用。近些年来,卵巢癌的治疗活动中监测肿瘤的活动,一直以CA125、B超等来进行。近几年来国际上逐渐开始重视了溶血磷脂酸（LPA）在其间的价值。在卵巢癌的治疗活动中LPA和CA125的作用有多大的差别呢？能否使用LPA来替代CA125在卵巢癌治疗活动中的监测作用呢？本次研究就对此进行了一些简单的比对,希望给大家一个满意的结果。     

HER receptor signaling confers resistance to the insulin-like growth factor-I receptor inhibitor, BMS-536924

We have reported previously the activity of the insulin-like growth factor-I (IGF-IR)/insulin receptor (InsR) inhibitor, BMS-554417, in breast and ovarian cancer cell lines. Further studies indicated treatment of OV202 ovarian cancer cells with BMS-554417 increased phosphorylation of HER-2. In addition, treatment with the pan-HER inhibitor, BMS-599626, resulted in increased phosphorylation of IGF-IR, suggesting a reciprocal cross-talk mechanism. In a panel of five ovarian cancer cell lines, simultaneous treatment with the IGF-IR/InsR inhibitor, BMS-536924 and BMS-599626, resulted in a synergistic antiproliferative effect. Furthermore, combination therapy decreased AKT and extracellular signal-regulated kinase activation and increased biochemical and nuclear morphologic changes consistent with apoptosis compared with either agent alone. In response to treatment with BMS-536924, increased expression and activation of various members of the HER family of receptors were seen in all five ovarian cancer cell lines, suggesting that inhibition of IGF-IR/InsR results in adaptive up-regulation of the HER pathway. Using MCF-7 breast cancer cell variants that overexpressed HER-1 or HER-2, we then tested the hypothesis that HER receptor expression is sufficient to confer resistance to IGF-IR-targeted therapy. In the presence of activating ligands epidermal growth factor or heregulin, respectively, MCF-7 cells expressing HER-1 or HER-2 were resistant to BMS-536924 as determined in a proliferation and clonogenic assay. These data suggested that simultaneous treatment with inhibitors of the IGF-I and HER family of receptors may be an effective strategy for clinical investigations of IGF-IR inhibitors in breast and ovarian cancer and that targeting HER-1 and HER-2 may overcome clinical resistance to IGF-IR inhibitors.     

Sensitivity to pertuzumab (2C4) in ovarian cancer models: cross-talk with estrogen receptor signaling

Pertuzumab (Omnitarg, rhuMab 2C4) is a humanized monoclonal antibody, which inhibits HER2 dimerization. Because it has shown some clinical activity in ovarian cancer, this study sought to identify predictors of response to this agent in a model of ovarian cancer. A panel of 13 ovarian cancer cell lines was treated with heregulin beta1 (HRGbeta1) or transforming growth factor-alpha, and cell proliferation was assessed. Both agents increased cell number in the majority of cell lines studied, the response to both being similar (r = 0.83; P = 0.0004, Pearson test). HRGbeta1 stimulation could be partially reversed by pertuzumab in 6 of 13 cell lines, with complete reversal in PE04 and PE06 cells. Addition of pertuzumab to transforming growth factor-alpha-stimulated cells produced growth inhibition in 3 of 13 cell lines (PE01, PE04, and PE06). The magnitude of HRGbeta1-driven growth stimulation correlated significantly with an increase in extracellular signal-regulated kinase 2 (P = 0.037) but not Akt (P = 0.99) phosphorylation. Such HRGbeta1-driven phosphorylation of extracellular signal-regulated kinase 1/2 and Akt could be reduced with pertuzumab, accompanied by changes in cell cycle distribution. In cell lines responsive to pertuzumab, HRGbeta1-enhanced phosphorylation of HER2 (Tyr(877)) was reduced. Estrogen-stimulated changes in growth, cell cycle distribution, and signaling were reversed by pertuzumab, indicating cross-talk between HER2 and estrogen signaling. These data indicate that there is a subset of ovarian cancer cell lines sensitive to pertuzumab and suggest possible predictors of response to identify patients who could benefit from this therapy. Furthermore, we have identified an interaction between HER2 and estrogen signaling in this disease.     

Sensitization of breast cancer cells to radiation by trastuzumab

HER2, a member of the human epidermal growth factor (EGF) receptor family, not only plays important roles in the progression of breast cancer tumorigenesis and metastasis, but may protect cancer cells from conventional cytotoxic therapies as well. In the current study, we evaluated the effect of targeting HER2 on radiosensitization of human breast cancer cells. Using six breast cancer cell lines with various levels of HER2 (BT474, SKBR3, MDA453, MCF7, ZR75B, and MDA468), we found that trastuzumab (Herceptin), a humanized monoclonal antibody that may inhibit breast cancer cell proliferation but does not induce apoptosis when used alone, enhanced radiation-induced apoptosis of the cells in a HER2 level-dependent manner. We furthered this study in MCF7 cells transfected for high levels of HER2 (MCF7HER2). Compared with parental or control vector-transfected MCF7 cells, MCF7HER2 cells showed increased phosphorylation of at least two important HER2 downstream molecules, protein kinase B/Akt and mitogen-activated protein kinase (MAPK), and increased resistance to radiotherapy, as shown by reduced induction of apoptosis and increased cell clonogenic survival after radiation. Exposure of the cells to trastuzumab down-regulated the levels of HER2 and reduced phosphorylation levels of Akt and MAPK in MCF7HER2 cells, and sensitized these cells to radiotherapy. When specific inhibitors of the phosphatidylinositol 3-kinase (PI3-K) and MAPK kinase (MEK) pathways were used, we found that exposure of MCF7HER2 cells to the PI3-K inhibitor LY294002 inhibited Akt phosphorylation and radiosensitized the cells, whereas the radiosensitization effect by the MEK inhibitor PD98059 was relatively weaker, albeit the phosphorylation of MAPK was reduced by PD98059 treatment. Our results indicate that the PI3-K pathway might be the major pathway for trastuzumab-mediated radiosensitization of breast cancer cells.     

Platelet-derived lysophosphatidic acid supports the progression of osteolytic bone metastases in breast cancer

The role of lysophosphatidic acid (LPA) in cancer is poorly understood. Here we provide evidence for a role of LPA in the progression of breast cancer bone metastases. LPA receptors LPA(1), LPA(2), and LPA(3) were expressed in human primary breast tumors and a series of human breast cancer cell lines. The inducible overexpression of LPA(1) in MDA-BO2 breast cancer cells specifically sensitized these cells to the mitogenic action of LPA in vitro. In vivo, LPA(1) overexpression in MDA-BO2 cells enhanced the growth of subcutaneous tumor xenografts and promoted bone metastasis formation in mice by increasing both skeletal tumor growth and bone destruction. This suggested that endogenous LPA was produced in the tumor microenvironment. However, MDA-BO2 cells or transfectants did not produce LPA. Instead, they induced the release of LPA from activated platelets which, in turn, promoted tumor cell proliferation and the LPA(1)-dependent secretion of IL-6 and IL-8, 2 potent bone resorption stimulators. Moreover, platelet-derived LPA deprivation in mice, achieved by treatment with the platelet antagonist Integrilin, inhibited the progression of bone metastases caused by parental and LPA(1)-overexpressing MDA-BO2 cells and reduced the progression of osteolytic lesions in mice bearing CHO-beta3wt ovarian cancer cells. Overall, our data suggest that, at the bone metastatic site, tumor cells stimulate the production of LPA from activated platelets, which enhances both tumor growth and cytokine-mediated bone destruction.     

Polyclonal antibodies against gp185HER2 peptides: their putative role in the identification of a particular HER2 status in patients with breast cancer

The HER2 oncogene and its relative oncoprotein, gp185HER2, a transmembrane glycoprotein belonging to the epidermal growth factor receptor family, are overexpressed in a wide range of solid tumors including breast and ovarian cancer. In patients with breast cancer, both humoral and cell-mediated HER2 immune responses have been found as well as in some patients with gp185HER2 nonoverexpressing tumors. To establish whether peptide sequences identified as HLA-A2-restricted T-cell epitopes are expressed in breast tumor cell lines and tissues, we produced and characterized by different methodologic approaches polyclonal antibodies raised against four gp185HER2 peptides. Two of the antibodies recognized peptides eluted from the HLA-A2 groove of the mDAmB231 breast cancer cell line expressing a basal level of gp185HER2. Paraffin-embedded primary and metastatic breast tumors were specifically immunostained by all four reagents, thereby showing an overlapping reactivity. When this immunoreactivity was compared with that obtained using two different monoclonal antibodies, in 105 breast primary tumors and 36 corresponding lymph node metastases, we identified a subset of tumors that were negative with anti-gp185HER2 monoclonal antibodies and positive with the four antipeptide antibodies. Our novel observations provide in vivo evidence of the complexity involved in evaluating HER2 expression, and open a new path for understanding the biologic significance of HER2 status in breast tumors.     

溶血磷脂酸甲胎蛋白等联检在卵巢癌早期诊断中的意义

目的探讨溶血磷脂酸（LPA）、血清肿瘤相关抗原125（CA125）、甲胎蛋白（AFP）含量对卵巢癌早期诊断的临床价值。方法早期卵巢癌（Ⅰ、Ⅱ期）患者36例,卵巢良性肿瘤36例,健康妇女36例。用定磷法测定LPA的含量,用电化学发光免疫法（ECLIA）测定CA125、AFP的含量。结果与良性肿瘤组和健康对照组相比,卵巢癌患者的LPA、CA125、AFP含量明显升高（P〈O．001）;以单一阳性指标作为标准,对卵巢癌的阳性率分别为86％、67％、31％;特异性分别为94％、75％、56％;联合检验上述3项肿瘤标志物阳性率为92％,特异性为97％。结论联合检测LPA、CA125、AFP对卵巢癌的早期诊断有着较高的临床应用价值。     

乳腺癌HER-2靶向治疗的现状与发展前景

目的人表皮生长因子受体2(HER2)是表皮生长因子受体(EGFR或Er Bb)家族中扮演中心角色的一个成员。本文对HER2靶向治疗的现状与发展方向作一综述。方法查阅Pubmed、中国知网、万方、维普等数据库与乳腺癌HER2靶向治疗方面相关的文章,共纳入52篇文献。结果 HER2信号能够调节细胞增殖、存活和分化,与乳腺癌、胃癌、肺癌与卵巢癌等多种肿瘤的发生发展密切相关。目前使用的HER2靶向治疗有助于改善临床化疗药物疗效、提高患者无疾病生存期和5年存活率,但肿瘤的耐药性也越来越严重,已经引起人们的关注。结论 HER2靶向治疗对提高HER2阳性乳腺癌患者治疗效果起到重要作用,其耐药性需要进一步研究解决。