高特异性HER-2/neu肽疫苗的实验研究

目的　设计并制备显著激活细胞毒性 T细胞 (CTL)的高特异性 HER- 2 / neu肽疫苗 ,并对其进行活性实验研究。方法　根据 T site蛋白质分析软件模拟的 HER- 2 / neu空间构象 ,选取、合成 2种与 MHC- I类分子高亲和性的区段 P4 2 - 50 、P782 - 790 并对其进行修饰。建立 BALB/ c小鼠的乳腺癌模型 ,用上述多肽作疫苗、GM- CSF做佐剂 ,免疫小鼠 ,分别应用 MTT法、EL ISPOT法、颗粒酶释放法测定 T细胞的增殖 ,T细胞分泌 IFN-γ,CTL对肿瘤细胞的杀伤活性。结果　经修饰的 HER- 2 / neu多肽疫苗可促进 T细胞增殖 ,促进 T细胞分泌 IFN-γ及 CTL杀伤乳腺癌细胞。结论　经修饰过的 HER- 2 / neu多肽疫苗可显著激活细胞免疫 ,增加对乳腺癌细胞的杀伤作用。     

HER2／neu在胃癌中的研究进展

HER2/neu与多种肿瘤的发生、发展关系密切。它通过下游的信号转导途径参与肿瘤细胞的增殖、凋亡、浸润与侵袭等基因的调控。HER2/neu在胃癌中存在高表达或基因扩增现象,并可能影响胃癌患者的预后,也可能参与了胃癌肝转移过程。HER2/neu有望成为胃癌治疗的一个新靶点。     

RNAi抑制前列腺癌PC-3M细胞HER-2/neu基因表达的研究

目的研究利用RNA干涉技术（RNAi）抑制前列腺癌细胞PC-3 M中HER-2/neu基因表达的方法.方法用DNA重组技术构建携带与短的发夹结构干涉RNA相对应DNA序列的pSUPER-HER-2重组质粒,采用脂质体将pSUPER-HER-2重组质粒稳定转染至PC-3M细胞,流式细胞技术（FCM）分析PC-3 M细胞中HER-2蛋白的表达情况,RT-PCR鉴定HER-2 mRNA的表达情况.结果获得了用RNAi稳定抑制HER-2/neu基因表达的细胞株PC-3 MpSUPER-HER-2,该细胞株中的HER-2/neu mRNA及HER-2/neu蛋白的表达均显著降低.结论成功构建pSUPER-HER-2/neu重组质粒,并能有效抑制PC-3 M细胞中HER-2/neu表达.     

抗HER2乳腺癌ADC药物研究进展

<正>根据世界癌症发病率统计,乳腺癌已超越肺癌成为女性中发病率居于首位的癌症,尽管乳腺癌的诊断方法及综合治疗手段近几年来不断发展和革新,死亡率仍处于女性恶性肿瘤第二的高位[1]。人表皮生长因子受体(HER)2异常表达的患者占所有乳腺癌的15%～20%[2],由于HER2具有强大的催化激酶活性进而触发许多不同的下游途径,如磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)和膜受体酪氨酸蛋白激酶(MAPK)途径[3,4]。以曲妥珠单抗为基础,     

重组人HER2/neu ICD在毕赤酵母中的表达及鉴定

目的 用毕赤酵母表达体系制备具有与天然HER2/neu ICD相同结构和活性的重组HER2/neu ICD.方法 PCR法自质粒pCMV中克隆HER2/neu ICD基因,构建真核表达载体pPICZα/HER2/neu ICD,电转化至毕赤酵母菌株X-33.对基因转染Zeocin抗性阳性的克隆,采用PCR法筛选出稳定表达HER2/neu ICD的菌株.最后用SDS-PAGE鉴定甲醇诱导的培养上清液中的HER2/neu ICD.结果 构建pPICZα/HER2/neu ICD真核表达载体并经电转化获得Zeocin抗性阳性的克隆;PCR法筛选了稳定表达HER2/neu ICD的菌株;SDS-PAGE分析显示甲醇诱导后表达上清中含有HER2/neu ICD,分子量约为84 kDa.结论 成功构建pPICZα/HER2/neu ICD真核表达载体并筛选出稳定表达HER2/neu ICD的毕赤酵母工程菌.     

青年与老年乳腺癌患者ER、PR、HER2基因表达及预后的对比分析

我国乳腺癌发病率以每年2％ ～3％的速度增长,好发于40 ～ 50岁女性[1].我国流行病学调查显示,由于环境污染、不良生活习惯及人口老龄化等导致青年(≤35岁)及老年(≥65岁)[2]乳腺癌发病率升高[3,4].肿瘤流行病学调查发现青年乳腺癌患者生物学行为及预后与老年患者截然不同,其中在雌激素受体(ER)、孕激素受体(PR)、人表皮生长因子受体(HER)2基因表达方面尤为突出,这也是这两个群体治疗策略的根本不同所在.现用SP法检测乳腺癌ER、PR表达、FISH法检测HER2状态[2009年NCCN指南指出荧光原位杂交(FISH)是检测HER2状态的金标准],总结≤35岁和≥65岁的乳腺癌患者发病情况、ER、PR、HER2表达状态的差异性及预后状况.     

体内外检测乙型肝炎DNA疫苗诱生小鼠特异性CTL活性的比较研究

目的：了解体内外两种方法检测乙型肝炎DNA疫苗诱生BALB／c小鼠特异性细胞毒性T淋巴细胞（CTL）活性的特点，比较其异同，建立与体外法互为补充，且更加直观、简便的活体CTL活性检测模型。方法：用HBV-S真核表达质粒pVAX1／S对SP2／0细胞进行转染，经鉴定后作为目的靶细胞（稳定转染并表达HBV主蛋白，命名为SP2／0-S细胞）；对照靶细胞为稳定转染pVAX1空载质粒的细胞（命名为SP2／0-P细胞）备用。活体诱生实验：用目的或对照靶细胞分别使小鼠荷瘤，观察免疫及非免疫小鼠在各靶细胞负荷下的生存时间与肿瘤的生长情况，设置不同组合做同期对照观察，小鼠生存率的差异可以提示体内特异性CTL是否存在。体外诱生实验：采用经典CTL活性检测方法，LDH释放法成品药盒。SP2／0-S细胞与SP2／0-P细胞作为靶细胞，免疫鼠与非免疫鼠的脾细胞为效应细胞，按照不同效／靶比共孵育后以酶标仪检测LDH的释放活性。结果：活体诱生实验：免疫小鼠存活时间明显长于对照小鼠（P〈0．05）。体外诱生实验：免疫鼠CTL活性在彬靶比为50／1时出现最大杀伤活性，为36．9％，明显优于对照组（P〈0．05）。结论：体内外两种方法均能够检测到乙型肝炎DNA疫苗诱生BALB／c小鼠特异性CTL活性，实验中发现体外LDH释放法CTL活性并不高，但是总体上仍然能够反映CTL活性。而活体诱生实验操作相对简便，表现更为直观，可望成为与经典方法相互补充的新型实验模型。     

HER-2/neu基因在非小细胞肺癌中的表达及其临床意义

目的探讨HER-2/neu基因在非小细胞肺癌中的表达及其临床意义。方法应用免疫组化法检测90份非小细胞肺癌组织及20份正常肺组织HER-2/neu基因的表达。结果 HER-2/neu基因在90份非小细胞肺癌组织中过度表达率为32.2%(29/90),20份正常肺组织中无过度表达。非小细胞肺癌的HER-2/neu基因的过度表达与年龄、性别、分化程度均无相关性(P均>0.05),与肿瘤的病理类型、淋巴结转移、临床分期、生存期相关(P均<0.05)。结论检测非小细胞肺癌组织中HER-2/neu基因的表达,有助于判断其预后,也为抗HER-2/neu基因的分子靶向治疗应用于非小细胞肺癌中提供理论依据。     

IL-2基因转导小鼠肝癌细胞的实验研究

用重组人白细胞介素-2(IL-2)含信号肽cDNA的缺陷型逆转录病毒,将IL-2基因转导入小鼠H_(22)肝癌细胞株,经PCR和RT-PCR检测证实了转导后的细胞DNA内确有NeoR和IL-2基因的存在的表达。分别用光镜和电镜观察了经IL-2基因转录的H_(22)细胞体外形态,MTT法检测细胞体外增殖能力以及皮下接种H_(22)-IL-2细胞后观察肿瘤生长能力。结果表明经IL-2基因转导的鼠肝癌细胞体外增殖能力没有明显变化,但体内致瘤性却显著下降,丝裂霉素处理后作为瘤苗接种能抑制其后野生型H_(22)细胞的再次攻击。本研究为制备新型瘤苗打下了基础。     

HER2阳性乳腺癌靶向治疗的研究进展

［摘要］　乳腺癌是全球女性发病率较高的恶性肿瘤,其中人类表皮生长因子受体2（HER2）阳性乳腺癌病例占总乳腺癌病例的20%～30%。与其他分子分型的乳腺癌比,HER2阳性乳腺癌预后更差,更易复发和转移。近年来,HER2阳性乳腺癌治疗备受关注,针对抗HER2的靶向药物能显著改善HER2阳性乳腺癌患者的疗效。该文对HER2阳性乳腺癌靶向治疗的研究进展作一综述。     

Pten loss promotes MAPK pathway dependency in HER2/neu breast carcinomas

Loss of the tumor suppressor gene PTEN is implicated in breast cancer progression and resistance to targeted therapies, and is thought to promote tumorigenesis by activating PI3K signaling. In a transgenic model of breast cancer, Pten suppression using a tetracycline-regulatable short hairpin (sh)RNA cooperates with human epidermal growth factor receptor 2 (HER2/neu), leading to aggressive and metastatic disease with elevated signaling through PI3K and, surprisingly, the mitogen-activated protein kinase (MAPK) pathway. Restoring Pten function is sufficient to down-regulate both PI3K and MAPK signaling and triggers dramatic tumor regression. Pharmacologic inhibition of MAPK signaling produces similar effects to Pten restoration, suggesting that the MAPK pathway contributes to the maintenance of advanced breast cancers harboring Pten loss.The PTEN (phosphatase and tensin homolog) tumor suppressor gene is mutated or silenced in a wide range of tumor types (1). PTEN encodes a phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase that counters the action of the phosphatidylinositol 3-kinases (PI3Ks), which otherwise transmit growth factor signals from receptor tyrosine kinases to downstream mediators such as the AKT family of serine/threonine-specific protein kinases (2). AKT, in turn, activates a series of downstream effectors that promote cellular proliferation and survival. Consequently, PTEN loss leads to hyperactivation of the PI3K pathway, and it is widely believed that this is the primary mechanism whereby PTEN loss drives tumorigenesis (3). Although cross-talk and feedback signaling makes the situation more complex (4), this molecular framework provides a strong rationale to target PI3K pathway components in PTEN-deficient tumors, and indeed, a variety of small-molecule antagonists with such activities are currently in clinical trials (5, 6).Deregulation of the PI3K pathway is common in breast cancer and most frequently occurs through mutations in phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) (7). By contrast, PTEN mutation or loss is less frequent at diagnosis but instead is associated with disease progression (8, 9). For example, PTEN inactivation often arises in oncogenic receptor tyrosine kinase human epidermal growth factor receptor 2 (HER2/neu) amplified tumors in patients who acquire resistance to the HER2/neu targeting agent trastuzumab (10–14). Similarly, PTEN mutations were recently reported in a patient harboring PIK3CA mutations that developed resistance to the PI3Kα inhibitor BYL719 (15). Thus, PTEN inactivation occurs in advanced disease in patients with poor prognosis, defining a breast cancer subtype for which there is an unmet clinical need.Studies using mouse models have confirmed the importance of PI3K signaling in breast cancer (16). Transgenic mice that overexpress mutant PIK3CA in conjunction with HER2/neu recapitulate resistance to anti-HER2/neu therapies (17), and conditionally overexpressed mutant PIK3CA in the mammary gland gives rise to tumors at long latency that regress upon oncogene withdrawal (18). Although these observations contribute to the rationale for targeting PI3K pathway components in breast cancer, they use a model in which mutant PIK3CA is expressed at unphysiological levels and serves as the initiating event. Furthermore, studies using conditional knockout mice indicate that deregulation of the endogenous PI3Ks indirectly through Pten inactivation can promote advanced disease in combination with HER2/neu (19, 20). Still, whether sustained PTEN inactivation is needed for the maintenance of advanced cancers remains unknown. Resolving this issue may reveal cellular dependencies and, as such, instruct the clinical use of molecularly targeted agents attacking the PTEN network. In this study, we explore the impact of genetic and pharmacologic manipulation of the Pten pathway in breast cancer. Unexpectedly, our results reveal that Pten loss is required to maintain advanced disease by enhancing signaling through both the PI3K and mitogen-activated protein kinase (MAPK) cascades.     

The epidemiology of Her-2/neu and P53 in breast cancer

Breast cancer is an etiologically heterogeneous disease with marked geographical variations. Joint consideration of the relationship between specific molecular alterations and known or suspected epidemiologic risk factors for this disease should help distinguish subgroups of women that are at elevated risk of developing breast cancer. In this article, we present a comprehensive literature review of the etiologic and prognostic roles of Her-2/neu and P53 among women. In addition, we discuss the advantages and limitations of using biomarkers in epidemiological studies. We conclude that more research is needed to understand the complex relationships between genetic alterations and etiologic risk factors for breast cancer.     

Cardiotoxicity in HER2-positive breast cancer patients

Heart Failure Reviews - Due to the recent advances in diagnosis and management of patients with HER2-positive breast cancer, especially through novel HER2-targeted agents, cardiotoxicity becomes an...     

早期HR阳性/HER2阳性乳腺癌的治疗选择与思考

［摘要］　激素受体（HR）阳性/人表皮生长因子受体2（HER2）阳性乳腺癌在分子功能、生物过程、信号通路、临床行为、治疗敏感性和内在生物学方面与其他分子亚型乳腺癌存在差异,因此被认为是乳腺癌分子分型中较为特殊的一类。目前,HR阳性/HER2阳性乳腺癌在治疗策略的选择上存在较多的争议和不确定性。如何选择疗效更好、安全性更高、可及性更强的治疗策略,是目前及将来较长一段时间内需要探索的问题。该文将通过新辅助和辅助治疗阶段多个临床研究中HR阳性/HER2阳性乳腺癌患者的相关数据进行分析,从数据中找寻HR阳性/HER2阳性乳腺癌患者的特殊性,为制定更适合的个体化治疗策略提供依据。