Mechanism of lapatinib-induced cell tolerance from the perspective of metabolic changes in ErbB2 positive breast cancer cells

目的研究酪氨酸激酶受体ErbB2/ErbB1的小分子抑制剂lapatinib（GW2974）对ErbB2/ErbB1阳性乳腺癌细胞代谢的影响，探讨lapatinib对代谢的调控机制。为揭示lapatinib对ErbB2/ErbB1阳性乳腺癌细胞凋亡诱导与细胞耐受机制提供一定的研究依据，并为寻找新的联合给药靶点与方案克服其耐药现状提供积极线索。 方法实验选择乳腺癌细胞株BT474（ErbB2+）、SKBR3（ErbB2+）与MDA-MB-231（ErbB1+），设置Lapatinib浓度梯度与时间梯度组。利用Real-timePCR检测mRNA表达；WesternBlot方式检测代谢调控信号通...Objective: To investigate the effect of lapatinib on the metabolism of ErbB1/ErbB2 positive breast cancer cells, a small molecule inhibitor of the tyrosine kinase receptor of ErbB2/ErbB1, and to explore the metabolic regulation mechanisms of lapatinib effects. At the same time, to provide insight to reveal the mechnism of the apoptosis lapatinib induced of ErbB2/ErbB1 positive breast cancer cells...学位：理学硕士院系专业：医学院_药理学学号：2452011115334

High-precision measurements of Fe and Zn isotopic ratios in marine sediments

介绍海洋沉积物fE和zn同位素化学前处理及测定方法,报道南海西部夏季上升流区两个沉积物柱样的fE和zn同位素组成。样品采用Hf+HnO3+HClO4常压消解,经脱盐后,转化为氯化物形式并经离子交换柱分离纯化后,用多接收器等离子体质谱法测定fE和zn同位素比值。该前处理方法可以快捷地实现海洋沉积物的消解、有机质的去除和海盐脱离;结合相关测试流程,可获得较高的δ56fE(0.10‰,2Sd)和δ66zn分析精度(0.11‰,2Sd)。两个沉积物柱样的δ56fE值(相对于IrMM-014)和δ66zn值(相对于JMC3-0749C)随深度变化不明显,两柱之间也无明显差异。总体上,南海西部上升流区1~2 kA以来的沉积物δ56fE值(0.04‰~0.20‰)和δ66zn值(0.12‰~0.30‰)与已报道的黄土和气溶胶、火成岩以及大部分海洋沉积物接近,明显高于静海相海洋沉积物的δ56fE值。We present an optimized chemical separation and purification procedure as well as an analytical method for measuring Fe and Zn isotopic composition in marine sediments.We also report Fe and Zn isotopic compositions in two sediment cores with an age of 1-2 ka from the summer upwelling region in the western South China Sea.Samples were dissolved with an acid solution(HF + HNO3 + HClO4) under normal pressure,followed by desalination and transformation to chloride matrix.After separation and purification,Fe and Zn isotopic ratios were determined with a Nu Plasma HR MC-ICPMS.Results demonstrate that this chemical preparation can rapidly dissolve marine sediments,remove organic matter,and,in combination with this analytical technique,we achieved high analytical precisions of 0.10‰ for δ56Fe(2SD) and of 0.11‰ for δ66Zn(2SD).These two sediment cores had nearly invariant δ56Fe and δ66Zn values throughout the sediment cores.Moreover,δ56Fe(0.04‰-0.20‰) and δ66Zn(0.12‰-0.30‰) from these 1-2 ka marine sediments are overall close to those reported for loesses and aerosols,igneous rocks and other marine sediments worldwide,but much higher than those deposited in euxinic environments.国家自然科学基金创新群体项目(40921001;40521003);国土资源部公益性行业研究专项经费项目(201011027

酪氨酸激酶抑制药GW2974诱导乳腺癌细胞BT474耐药的代谢机制研究

目的通过比较酪氨酸激酶抑制药GW2974给药后母本细胞株BT474与耐药株r BT474之间代谢相关因子mRNA表达的差异,探讨r BT474可能的耐药机制。方法收集对数生长期r BT474和BT474细胞,接种于96孔板,加入含有GW2974的培养基,分别于孵育0,12,24和48 h后,通过噻唑蓝法检测BT474与r BT474细胞的增殖情况。用逆转录实时定量-聚合酶链反应检测12种相关代谢因子葡萄糖转运体4(GLUT4)、2,6-二磷酸果糖激酶(PFK-2)、丙酮酸激酶2(PKM2)、乳酸脱氢酶(LDHA)、丙酮酸羧化酶(PC)、6-磷酸果糖激酶(G-6-PD)、脂肪酸合成酶(FASN)、脂酰肉碱转移酶1 (CPT1A)、葡萄糖调节蛋白75(GRP75)、电压依赖性阴离子通道蛋白1 (VDAC1)、葡萄糖调节蛋白78(GRP78)和钙网蛋白(CALR)的mRNA在BT474细胞和r BT474细胞中给药前后的表达水平。结果给予GW2974后,母本细胞BT474中糖脂代谢相关因子GLUT4、PFK-2、PKM2、LDHA、PC、G-6-PD、FASN和CPT1A的mRNA表达水平均明显下调,线粒体应激因子GRP75、VDAC1与内质网应激因子CALR表达水平均明显下调,内质网应激因子GRP78表达明显上调。而在耐药株r BT474中给药后这12种代谢相关因子的mRNA表达水平均明显上调。结论耐药株r BT474可能通过调控糖脂代谢以及细胞应激等过程使细胞耐受性增强。福建省自然科学基金面上项目(2012J01417