In Vitro Effects of Calcium Fructoborate on fMLP-stimulated Human Neutrophil Granulocytes

Discovery of naturally occurring boron complexes with organic compounds containing hydroxyl groups, sugars, and polysaccharides, adenosine-5-phosphate, pyridoxine, riboflavin, dehydroascorbic acid, and pyridine nucleotides led to the reassessment of the biochemical role of boron. Boron’s anti-inflammatory actions were claimed but not yet demonstrated. This study investigated the effects of calcium fructoborate (CF) on the human polymorphonuclear neutrophils (PMN) that play a central role in the inflammatory response. Our results demonstrated that CF exposure induced a dose-dependent decrease in cell viability. Treatment of PMN cells, for 24 h, with 22,500 μM CF led to a decrease in cell viability by 61.1%, an inhibition of respiratory burst by 92.9% in the case of fMLP-stimulated cells, a diminution of intracellular level of superoxide anion with 59.3%, and a stimulation of superoxide dismutase activity by 72% in unstimulated PMN cells. Altogether, these results suggest the antioxidant and anti-inflammatory properties of CF.     

Experimental and Theoretical Studies of Calcium Fructoborate

Calcium fructoborate samples of composition Ca(C₆H₁₀O₆BO)₂·3.5H₂O were characterized by chemical analysis, infrared and Raman spectroscopy, and thermoanalytical (thermogravimetric and differential thermal analysis) data. Theoretical studies, using density functional theory, were made for seven different structural models of the fructoborate moiety, and the most stable structure could be derived from these calculations. The results of the theoretical study also allow improving the assignment of the vibrational spectra of the compound.     

二甲双胍联合阿霉素应用对人乳腺癌细胞增殖和凋亡的影响

目的：观察二甲双胍联合阿霉素应用对人乳腺癌细胞MDA-MB-231增殖和凋亡的影响。方法：MTT法分别检测二甲双胍、阿霉素和二甲双胍联合阿霉素对MDA-MB-231细胞生长的抑制作用;平板克隆实验检测二甲双胍联合阿霉素对MDA-MB-231细胞克隆形成能力的影响;流式细胞仪检测二甲双胍联合阿霉素对MDA-MB-231细胞凋亡的影响。结果：二甲双胍和阿霉素分别对MDA-MB-231细胞生长有抑制作用,二甲双胍联合阿霉素应用对MDA-MB-231细胞生长的抑制作用更加显著,并且随着药物浓度的增加而增加;二甲双胍联合阿霉素应用与单药相比能够明显降低MDA-MB-231细胞克隆形成率,并且促进细胞凋亡。结论：二甲双胍联合阿霉素应用与单药相比能够显著抑制人乳腺癌细胞MDA-MB-231细胞的增值,促进其凋亡,可见两药联用对肿瘤细胞的杀伤具有协同性。     

Distinct Regulation of Cytoplasmic Calcium Signals and Cell Death Pathways by Different Plasma Membrane Calcium ATPase Isoforms in MDA-MB-231 Breast Cancer Cells

Plasma membrane calcium ATPases (PMCAs) actively extrude Ca(2+) from the cell and are essential components in maintaining intracellular Ca(2+) homeostasis. There are four PMCA isoforms (PMCA1-4), and alternative splicing of the PMCA genes creates a suite of calcium efflux pumps. The role of these different PMCA isoforms in the control of calcium-regulated cell death pathways and the significance of the expression of multiple isoforms of PMCA in the same cell type are not well understood. In these studies, we assessed the impact of PMCA1 and PMCA4 silencing on cytoplasmic free Ca(2+) signals and cell viability in MDA-MB-231 breast cancer cells. The PMCA1 isoform was the predominant regulator of global Ca(2+) signals in MDA-MB-231 cells. PMCA4 played only a minor role in the regulation of bulk cytosolic Ca(2+), which was more evident at higher Ca(2+) loads. Although PMCA1 or PMCA4 knockdown alone had no effect on MDA-MB-231 cell viability, silencing of these isoforms had distinct consequences on caspase-independent (ionomycin) and -dependent (ABT-263) cell death. PMCA1 knockdown augmented necrosis mediated by the Ca(2+) ionophore ionomycin, whereas apoptosis mediated by the Bcl-2 inhibitor ABT-263 was enhanced by PMCA4 silencing. PMCA4 silencing was also associated with an inhibition of NFκB nuclear translocation, and an NFκB inhibitor phenocopied the effects of PMCA4 silencing in promoting ABT-263-induced cell death. This study demonstrates distinct roles for PMCA1 and PMCA4 in the regulation of calcium signaling and cell death pathways despite the widespread distribution of these two isoforms. The targeting of some PMCA isoforms may enhance the effectiveness of therapies that act through the promotion of cell death pathways in cancer cells.     

鞘氨醇激酶1参与依托泊苷抑制人乳腺癌MDA-MB-231细胞增殖、迁移的研究

目的利用抑制乳腺癌MDA-MB-231细胞中SK-1基因表达,结合依托泊苷对细胞增殖的影响,研究乳腺癌的治疗新方法。方法将依托泊苷分别处理野生型及SK-1敲除型MDA-MB-231细胞,^3H-TdR掺入法分析细胞增殖,Transwell法分析细胞迁移,Western印迹检测SK-1蛋白表达及细胞周期检验点相关信号因子的蛋白表达,RT-PCR检测细胞内SK-1的mRNA表达量。结果依托泊苷在较高剂量时,MDA-MB-231细胞存活率明显下降,但依托泊苷却呈浓度依赖性促进乳腺癌细胞SK-1 mRNA及蛋白水平表达,将SK-1敲除,细胞迁移率下降,而且可以增强G1期各抑癌基因的激活或高表达,使细胞周期阻滞。结论SK-1基因敲除有效增强肿瘤细胞对化疗药物的敏感性。     

亲骨转移乳腺癌细胞MDA-MB-231BO成瘤性的初步研究

目的通过比较亲骨转移乳腺癌细胞（MDA-MB-231BO）和亲代乳腺癌细胞（MDA-MB-231）的生长曲线和致瘤性,初步探讨MDA-MB-231BO细胞的生物学特性。方法MTT法测定两种细胞的生长曲线,并将两种乳腺癌细胞接种于裸鼠腋窝处皮下,建立乳腺癌细胞异种移植瘤动物模型,30 d后处死裸鼠,肿瘤组织及相关脏器官做病理检查。结果MTT法测得MDA-MB-231BO细胞生长速率高于MDA-MB-231细胞。接种两种乳腺癌细胞的裸鼠均长出肿瘤,成瘤率为100%。病理检查符合人乳腺癌细胞特征,MDA-MB-231BO组瘤体体积明显大于MDA-MB-231组（P〈0.05）。结论MDA-MB-231BO细胞生长速率高于MDA-MB-231细胞,而且MDA-MB-231BO在裸鼠体内的致瘤性强于MDA-MB-231。     

微流控芯片对乳腺癌细胞MDA-MB-231的捕获及再培养研究

目的:利用实验室构建的微流控芯片对乳腺癌细胞(MDA-MB-231)进行捕获,提高捕获率并保证细胞活性,实现再培养。抗肿瘤药物阿霉素处理正常培养和再培养的细胞,分析细胞内的基因表达变化。方法:对微流控芯片进行基底修饰,利用MUC1抗原与抗体特异性结合捕获肿瘤细胞,优化捕获条件提高捕获率。对微流控芯片捕获的细胞进行分离、收集和再培养。用1μmol/L阿霉素对正常培养和再培养的细胞分别孵育24h,然后提取RNA并逆转录合成c DNA。选择乳腺癌细胞中高表达及与肿瘤转移相关的基因FN1、ITGA6和LAMB3设计引物,以c DNA为模板分别进行RT-PCR扩增,对琼脂糖凝胶电泳结果进行灰度分析。结果:经MUC1抗体修饰的微流控芯片能有效地捕获肿瘤细胞,捕获率达80%±3%,释放率约98%,细胞释放后存活率高实现再培养。阿霉素对正常培养和再培养的乳腺癌细胞中FN1、ITGA6和LAMB3的基因表达均有抑制作用。结论:MUC1抗体修饰的微流控芯片能有效捕获乳腺癌细胞并实现再培养,捕获前后细胞内基因表达无显著差异,均能产生药物敏感性。     

C-藻蓝蛋白对乳腺癌细胞的抗癌作用研究

C-藻蓝蛋白具有抗癌、抗氧化、抗炎活性等多种功能,然而其对乳腺癌细胞的抗癌作用及机制尚不明确。本研究应用不同浓度(0~500μg/mL)的C-藻蓝蛋白处理人乳腺癌细胞系MDA-MB-468。研究显示,C-藻蓝蛋白以剂量依赖性方式抑制MDA-MB-468细胞的增殖并降低细胞的菌落形成能力。C-藻蓝蛋白通过上调了Fas和cleaved-caspase 3的表达并下调Bcl-2的表达来诱导细胞凋亡。C-藻蓝蛋白处理以剂量依赖性方式显著降低COX-2的表达并抑制细胞迁移能力。C-藻环蛋白以剂量依赖性方式促进p38 MAPK和JNK的磷酸化,p38 MAPK和JNK抑制剂处理可显著抑制C-藻红蛋白诱导的细胞死亡。本研究表明C-藻蓝蛋白能够抑制三阴性乳腺癌MDA-MB-468细胞的增殖,促进细胞凋亡,抑制细胞迁移能力。C-藻蓝蛋白的抗癌机制可能与激活p38 MAPK和JNK信号传导有关。     

The Effects of Boric Acid and Gibberellic Acid on the Growth of Fiber of Phragmites cormmuhis Trin

Water-cultured young plants of Phragmites communis Trin. were used in this experiment. They were treated with boric acid and gibberellic acid, separately or in combination, by method of foliage spraying. The results are summarized as follows: 1. In comparation with the controls, the length of the fibre cells was remarkably increased following the treatment with boric acid at concentrations ranging from 1 to 200 μmol/l. The optimum concentration of boric acid was found to be 100 μmol/l. Gibberellic acid alone at the concentration of 50 μmol/l also enhanced the length of the fibre cells. However, the best result was obtained in the group treated with a combination of boric acid and gibmberellic acid at a ratio of 200:50 μmol/l. A maximum increasement of fibre length and a greater ratio of length to width were observed in this experimental group. 2. Microscopic study indicates that the increase in both the number of the fibre cells and ,the thickness of the fibre layer are apparent in histological preparation. 3. In addition, The authors have also found that the incorporation of H3-glucose into the fibre cell wall was greatly enhanced by such combination treatment. 4. In conclusion, it has been demonstrated clearly that the boric acid and the gibberellic acid, when used in combination, produces an augmentative effect on both the growth and the development of the fibre cells of Phragmites communis Trin.     

MicroRNA-221/222对乳腺癌MDA-MB-231/DOX细胞阿霉素耐药性的影响

目的:探讨微小RNA-221/222(miR-221/222)对乳腺癌MDA-MB-231/阿霉素(DOX)细胞DOX耐药性的影响。方法:采用脂质体法转染miR-221/222抑制物(miR-221/222 inhibitor)至MDA-MB-231/DOX细胞内(Inhibitor组),同时设立空白对照组和转染无关序列的阴性对照组,采用实时荧光定量PCR (qRT-PCR)检测MDA-MB-231细胞株及MDA-MB-231/DOX细胞株的miR-221/222表达水平及转染效率;CCK-8法检测转染48 h后MDA-MB-231/DOX细胞对DOX药物敏感性的变化;流式细胞术(FCM)检测转染MDA-MB-231/DOX细胞的细胞凋亡率;蛋白免疫印迹实验(WB)检测转染后MDA-MB-231/DOX细胞内促凋亡蛋白p53上调凋亡调控因子(PUMA),Bcl2蛋白修饰因子(BMF)以及细胞周期蛋白激酶抑制因子p27(p27Kip1)的表达情况。结果:MDA-MB-231/DOX细胞中的miR-221/222表达水平高于亲本MDA-MB-231细胞(P0.05);MDA-MB-231/DOX细胞转染miR-221/222 inhibitor 96 h后,miR-221/222的表达水平低于空白对照组和阴性对照组(P0.05);与空白对照组相比,MDA-MB-231/DOX细胞转染miR-221/222 inhibitor 48h后,DOX继续处理48 h后,细胞的凋亡率明显升高,且细胞内的促凋亡蛋白PUMA,BMF以及p27Kip1的表达均增加(P0.05);DOX对inhibitor组耐药细胞的半数抑制浓度(IC50)显著低于空白对照组细胞及阴性对照组(P0.05)。结论:miR-221/222能够增加MDA-MB-231/DOX细胞对DOX的耐药性,这可能与下调促凋亡蛋白的表达有关;降低miR-221/222水平可诱导MDA-MB-231/DOX凋亡,并且上调促凋亡蛋白的表达,从而部分逆转MDA-MB-231/DOX对DOX的耐药性。     

UHRF1基因在乳腺癌循环肿瘤细胞中的表达研究

外周血液中乳腺循环癌肿瘤细胞的生物表征与转移性乳腺癌的严重程度密切相关,本研究的目的在于结合体外细胞实验探讨UHRF1基因对乳腺癌进展的意义。多重RNA原位分析乳腺癌循环肿瘤细胞(circulating tumor cells,CTCs)中UHRF1的表达;MTT法检测UHRF1基因转染对正常乳腺细胞增殖的影响;蛋白免疫印迹检测UHRF1基因转染对正常乳腺细胞中Bax蛋白和Bcl-2蛋白的影响;Caspase-3检测试剂盒检测正常乳腺细胞中Caspase-3活性;Transwell侵袭实验和划痕愈合实验检测UHRF1基因转染对正常乳腺细胞侵袭和迁移能力的影响。研究发现,UHRF1 RNA水平在乳腺癌循环肿瘤细胞中高表达;UHRF1基因增加正常乳腺细胞增殖率;UHRF1基因降低正常乳腺细胞中Caspase-3活性;UHRF1基因降低正常乳腺细胞中Bax蛋白的表达,增加Bcl-2蛋白的表达;UHRF1基因增强正常乳腺细胞侵袭和迁移能力。本研究初步说明,UHRF1可促进正常乳腺细胞增殖,抑制正常乳腺细胞凋亡,增强正常乳腺细胞侵袭和迁移能力。     

丙戊酸协同顺铂抑制乳腺癌和结直肠癌细胞生长

摘要 目的：探究丙戊酸（Valproic acid, VPA）协同顺铂抑制乳腺癌和结直肠癌细胞增殖。方法：首先使用Western blot 检测 VPA 对Acetyl-Histone H3蛋白水平的影响,使用Cell Counting Kit-8（CCK-8）法检测 VPA 对乳腺癌和结直肠癌细胞的细胞活力的影响。其次单药顺铂、VPA 和联合用药处理乳腺癌细胞 MDA-MB-231 和结直肠癌细胞 HCT-15,使用 IncuCyte 动态检测细胞生长过程和生长终点。结果：发现VPA 可抑制组蛋白去乙酰化酶的功能,升高Acetyl-Histone H3的蛋白水平,VPA 可抑制乳腺癌细胞和结直肠癌细胞增殖,且对 VPA 的药物敏感性相似;顺铂和 VPA 连用后可显著抑制乳腺癌和结直肠癌细胞增殖和活力。结论：本文发现 VPA 抑制组蛋白去乙酰化酶发挥抑制乳腺癌和结直肠癌细胞生长的新机制,并可以与顺铂连用提高抗肿瘤效果和药物敏感性,为同时患有癫痫和肿瘤的人群提供新的治疗思路。     

RNA干涉介导的Plk1沉默对乳腺癌细胞恶性生物表型的影响

目的：研究乳腺癌细胞中丝／苏氨酸蛋白激酶Plk1基因表达下调后对其恶性生物表型的影响。方法：利用pSitencer4．1-CMVneo质粒,分别构建针对Plk1基因的RNA干涉载体（pSilencer4．1-shPlk1）,利用脂质体Lipofectamine2000转染MCF-7细胞,G418筛选稳定的转染细胞系。半定量RT—PCR和Western blot分别检测Plk1基因mRNA和蛋白表达,MTT和克隆形成试验检测细胞增殖活性的变化,流式细胞仪分析细胞周期和凋亡的变化,最后分析MCF-7细胞对紫杉类药物（紫杉醇和多西他赛）化疗敏感性的变化。结果：成功筛选了稳定转染细胞系（MCF-7／shPlk1和MCF-7／shcontro1）。同MCF-7／shPlk1细胞相比,MCF-7／shPtkl细胞中Plk1基因mRNA和蛋白表达水平分别下调65．8％和74．4％（P〈0．05）。同MCF-7／shcontrol,MCF-7tshPlk1细胞增殖速度显著抑制,到第5天时抑制率达到44．9±3．2％（P〈0．05）。同时,MCF-7／shPlk1细胞的克隆形成能力显著降低（P〈0．01）流式细胞仪技术分析细胞周期结果表明：MCF-7／shPlk1细胞的G2／M期细胞比例显著增加了21．1±4．1％,而S期细胞比例则显著降低了（18．5±3．1％;P〈0．05）。流式细胞仪技术分析细胞凋亡结果表明：MCF-7／shPlk1细胞的凋亡率约显著增加了13．1±213％（P〈0．05）,同时还发现：MCF-7／shPlk1细胞中激活的caspase-3蛋白显著增加,Bcl-2蛋白显著降低,而Bax蛋白则显著增加。结论：RNA干涉载体能特异性下调乳腺癌细胞中Plk1基因的表达,从而抑制乳腺癌细胞的增殖和体外克隆形成能力,同时诱导乳腺癌细胞的G2／M期阻滞和细胞凋亡率显著增加。因此,靶向Plk1基因的生物治疗有望成为未来临床乳腺癌的一个重要的辅助治疗策略.     

The Role of Kif4A in Doxorubicin-Induced Apoptosis in Breast Cancer Cells

This study was to investigate the mechanism and role of Kif4A in doxorubicin-induced apoptosis in breast cancer. Using two human breast cancer cell lines MCF-7 (with wild-type p53) and MDA-MB-231 (with mutant p53), we quantitated the expression levels of kinesin super-family protein 4A (Kif4A) and poly (ADP-ribose) Polymerase-1 (PARP-1) by Western blot after doxorubicin treatment and examined the apoptosis by flow cytometry after treatment with doxorubicin and PARP-1 inhibitor, 3-Aminobenzamide (3-ABA). Our results showed that doxorubicin treatment could induce the apoptosis of MCF-7 and MDA-MB-231 cells, the down-regulation of Kif4A and upregulation of poly(ADP-ribose) (PAR). The activity of PARP-1 or PARP-1 activation was significantly elevated by doxorubicin treatment in dose- and time-dependent manners (P < 0.05), while doxorubicin treatment only slightly elevated the level of cleaved fragments of PARP-1 (P > 0.05). We further demonstrated that overexpression of Kif4A could reduce the level of PAR and significantly increase apoptosis. The effect of doxorubicin on apoptosis was more profound in MCF-7 cells compared with MDA-MB-231 cells (P < 0.05). Taken together, our results suggest that the novel role of Kif4A in doxorubicin-induced apoptosis in breast cancer cells is achieved by inhibiting the activity of PARP-1.     

Experimental Study on the Estrogen-Like Effect of Boric Acid

There are now considerable evidences that boric acid has reproductive and developmental toxicity, but it is uncertain whether such toxicity is caused by estrogen-like effect. Our objective is to determine the estrogen-like effect of boric acid. Proliferation assay of MCF-7 human breast cancer cells, uterotrophic assay, measure assay of the estradiol (E₂), proliferation assay of mucous membrane cells, and assay of estrogen receptor were conducted in this study. Boric acid could increase the weight of uterus of ovariectomized SD rats and the height of epithelium cells of mucous membrane, enhance the expression of the proliferating cell nucleus antigen, and reduce the density of estrogen receptors. However, boric acid could not affect the level of estradiol in serum and stimulate the proliferation of MCF-7 human breast cancer cells. In this study, boric acid exhibited the estrogen-like effect in vivo.     

Fatty Acid Synthase Mediates the Epithelial-Mesenchymal Transition of Breast Cancer Cells

This study aimed to investigate the role of fatty acid synthase (FASN) in the epithelial-mesenchymal transition (EMT) of breast cancer cells. MCF-7 cells and MCF-7 cells overexpressing mitogen-activated protein kinase 5 (MCF-7-MEK5) were used in this study. MCF-7-MEK5 cells showed stable EMT characterized by increased vimentin and decreased E-cadherin expression. An In vivo animal model was established using the orthotopic injection of MCF-7 or MCF-7-MEK5 cells. Real-time quantitative PCR and western blotting were used to detect the expression levels of FASN and its downstream proteins liver fatty acid-binding protein (L-FABP) and VEGF/VEGFR-2 in both in vitro and in vivo models (nude mouse tumor tissues). In MCF-7-MEK5 cells, significantly increased expression of FASN was associated with increased levels of L-FABP and VEGF/VEGFR-2. Cerulenin inhibited MCF-7-MEK5 cell migration and EMT, and reduced FASN expression and down-stream proteins L-FABP, VEGF, and VEGFR-2. MCF-7-MEK5 cells showed higher sensitivity to Cerulenin than MCF-7 cells. Immunofluorescence revealed an increase of co-localization of FASN with VEGF on the cell membrane and with L-FABP within MCF-7-MEK5 cells. Immunohistochemistry further showed that increased percentage of FASN-positive cells in the tumor tissue was associated with increased percentages of L-FABP- and VEGF-positive cells and the Cerulenin treatment could reverse the effect. Altogether, our results suggest that FASN is essential to EMT possibly through regulating L-FABP, VEGF and VEGFR-2. This study provides a theoretical basis and potential strategy for effective suppression of malignant cells with EMT.     

SOX7基因启动子甲基化水平对乳腺癌MDA-MB-231细胞株体外迁移和侵袭的影响

目的:探讨乳腺癌MDA-MB-231细胞中,Y性别决定区基因7(SOX7)基因启动子甲基化水平对细胞的体外迁移和侵袭的影响。方法:脂质体转染pcDNA3.0-DNA甲基转移酶3a(DNMT3a)质粒至MDA-MB-231细胞中,并于24h、48h及72h后,采用蛋白质免疫印迹实验(WB)检测细胞内DNMT3a蛋白表达水平;甲基化特异性定量PCR(Q-MSP)检测DNMT3a处理组、5-aza-C处理组及对照(Control)组MDA-MB-231细胞中的SOX7基因启动子DNA甲基化水平;实时荧光定量PCR(qRT-PCR)及WB实验检测各组MDA-MB-231细胞中的SOX7 m RNA和蛋白表达水平;细胞划痕实验及细胞侵袭实验检测各组MDA-MB-231细胞的迁移和侵袭能力。结果:pcDNA3.0-DNMT3a质粒转染MDA-MB-231细胞24h时,细胞内的DNMT3a蛋白表达水平最高。DNMT3a能够显著提高SOX7基因启动子DNA甲基化水平,而5-aza-C则抑制了SOX7基因启动子DNA甲基化水平(P0.05)。与Control组相比,DNMT3a处理组的MDA-MB-231细胞中,SOX7的m RNA及蛋白表达水平均明显下降,而5-aza-C处理组SOX7的m RNA及蛋白表达水平均明显增加(P0.05)。与Control组相比,DNMT3a处理组的MDA-MB-231细胞的迁移和侵袭能力均显著增强(P0.05),而5-aza-C处理组的MDA-MB-231细胞的迁移和侵袭能力变化不大(P0.05)。结论:在恶性肿瘤中,SOX7低表达表受其基因启动子高甲基化调节,且乳腺癌MDA-MB-231细胞中低表达的SOX7能够影响细胞的外迁移和侵袭能力。     

抑制c-FLIP基因促进TRAIL诱导的乳腺癌细胞凋亡(英文)

目的:探讨抑制c-FLIP的表达对TRAIL诱导乳腺癌细胞MCF-7凋亡的影响。方法:重组腺病毒Ad-c-FLIP-siRNA和Ad-sTRAIL单独及联合感染对TRAIL耐药的乳腺癌细胞MCF-7,应用实时荧光定量聚合酶链反应(Real-time PCR)检测病毒感染后各组细胞内c-FLIP和TRAIL的mRNA表达变化;MTT法和结晶紫染色法检测MCF-7细胞活性,Hoechst 33258荧光染色检测各组细胞的凋亡情况。结果:与阴性对照组比较,c-FLIP-siRNA组和c-FLIP-siRNA+TRAIL组c-FLIP的mRNA相对表达量分别是(0.32±0.16)和(0.39±0.48)倍;TRAIL组和c-FLIP-siRNA+TRAIL组TRAIL的mRNA相对表达量分别是(96.21±1.54)和(87.33±1.66)倍;TRAIL组、c-FLIP-siRNA组及c-FLIP-siRNA+TRAIL组的抑制率(%)分别为(60.27±1.25)、(11.34±1.74)及(74.91±2.12)。对比阴性对照组的凋亡率(3.12±1.54),TRAIL组(12.79±2.46)和c-FLIP-siRNA+TRAIL组(25.50±3.17)组的凋亡率明显增高(P0.05),c-FLIP-siRNA组(6.85±2.82)的凋亡率变化不明显,差异无统计学意义(P0.05)。结论:siRNA抑制c-FLIP基因的表达能显著促进TRAIL对乳腺癌细胞MCF-7凋亡的诱导作用。     

抑制c-FLIP基因促进TRAIL诱导的乳腺癌细胞凋亡

目的：探讨抑制c-FLIP的表达对TRAIL诱导乳腺癌细胞MCF-7凋亡的影响。方法：重组腺病毒Ad-c-FLIP-siRNA和Ad-sTRAIL单独及联合感染对TRAIL耐药的乳腺癌细胞MCF-7,应用实时荧光定量聚合酶链反应（Real-time PCR）检测病毒感染后各组细胞内c-FLIP和TRAIL的mRNA表达变化;MTT法和结晶紫染色法检测MCF-7细胞活性,Hoechst 33258荧光染色检测各组细胞的凋亡情况。结果：与阴性对照组比较,c-FLIP-siRNA组和c-FLIP-siRNA＋TRAIL组c-FLIP的mRNA相对表达量分别是（0.32±0.16）和（0.39±0.48）倍;TRAIL组和c-FLIP-siRNA＋TRAIL组TRAIL的mRNA相对表达量分别是（96.21±1.54）和（87.33±1.66）倍;TRAIL组、c-FLIP-siRNA组及c-FLIP-siRNA＋TRAIL组的抑制率（%）分别为（60.27±1.25）、（11.34±1.74）及（74.91±2.12）。对比阴性对照组的凋亡率（3.12±1.54）,TRAIL组（12.79±2.46）和c-FLIP-siRNA＋TRAIL组（25.50±3.17）组的凋亡率明显增高（P〈0.05）,c-FLIP-siRNA组（6.85±2.82）的凋亡率变化不明显,差异无统计学意义（P〉0.05）。结论：siRNA抑制c-FLIP基因的表达能显著促进TRAIL对乳腺癌细胞MCF-7凋亡的诱导作用。     

人乳腺癌MDA-MB-231细胞裸鼠移植瘤模型的建立及生物学特性研究

目的-建立人乳腺癌MDA-MB-231细胞株裸小鼠模型,研究其生物学特性,观察MDA-MB-231乳腺癌细胞在移植前后的形态学变化。方法-将人乳腺癌细胞MDA-MB-231接种于裸鼠腋窝处皮下,每3天测量肿瘤大小,第30天处死小鼠。肿瘤组织及相关脏器送病理切片。皮下肿瘤组织细胞及细胞株培养HE染色。结果-肿瘤生长较快,成功率为72%,病理检查符合人乳腺癌细胞特征。肿瘤组织细胞及培养细胞形态学未见显著差异。结论-人乳腺癌细胞株MDA-MB-231裸小鼠模型建立方法较简便,细胞形态无明显差异,且保持了人乳腺癌的生物学特性。