共查询到20条相似文献,搜索用时 15 毫秒
1.
目的:探讨RNA干扰技术沉默survivin基因表达对人乳腺癌细胞增殖和凋亡的影响.方法:设计并合成靶向survivin的小分子干扰RNA(siRNA),在脂质体(lipidosome)的介导下转染人乳腺癌细胞株MCF-7,荧光倒置显微镜观察细胞的转染效率; MTT(四甲基偶氮唑盐)比色法分析检测各组癌细胞的存活率;流式细胞计数检测各组癌细胞周期和凋亡指数;Western Blot方法观察对MCF-7细胞survivin mRNA和蛋白质表达的抑制效率.结果:Survivin-siRNA能有效封闭survivin基因的表达,使survivin的mRNA相对水平明显降低(P<0.05);Survivin-siRNA能明显抑制细胞增殖(P<0.05);Survivin-siRNA使细胞G2/M期细胞百分比明显增加,S期的细胞百分比显著减少(P<0.05).结论:利用RNA干扰技术沉默survivin基因的表达可以显著抑制MCF-7细胞的增殖,并在一定程度上诱导其自发凋亡,靶向survivin的RNA干扰技术在乳腺癌的基因治疗中具有一定的研究价值. 相似文献
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利用siRNA阻抑survivin基因表达诱导乳腺癌细胞系MCF-7细胞凋亡的研究 总被引:5,自引:0,他引:5
背景与目的:survivin属IAP基因家族成员,表达于多种肿瘤组织中,能促使细胞逃避凋亡,并促进细胞的异常有丝分裂。本研究旨在探讨敲除survivin基因后,癌细胞的增殖和凋亡情况。方法:利用RNAi阻抑人乳腺癌细胞内survivin基因的表达,RT-PCR和Westernblot法分析survivin基因mRNA和蛋白的表达,MTT法检测细胞生长增殖抑制率,流式细胞术检测细胞凋亡率。结果:survivinsiRNA转染组,survivin基因表达水平与未转染组比较下调了64%。随着siRNA浓度的增加,细胞增殖抑制率逐渐增高,200nmol/L剂量组细胞增殖抑制率最高,可达60.9%。不同浓度的siRNA可不同程度地诱导细胞凋亡,200nmol/L剂量组凋亡率最高,可达29.0%。结论:survivinsiRNA有可能成为治疗乳腺癌的新药物。 相似文献
3.
Differential regulation of growth and invasiveness of MCF-7 breast cancer cells by antiestrogens 总被引:6,自引:0,他引:6
E W Thompson R Reich T B Shima A Albini J Graf G R Martin R B Dickson M E Lippman 《Cancer research》1988,48(23):6764-6768
Estrogen increases the ability of the estrogen-dependent MCF-7 human breast cancer cell line to both proliferate and invade through an artificial basement membrane. In studying the response of MCF-7 cells to various antiestrogens, we found that 4-hydroxytamoxifen and tamoxifen inhibited cell proliferation but increased their invasiveness. In contrast, the structurally unrelated benzothiophene antiestrogens, LY117018 and LY156758, were potent antiproliferative agents which did not stimulate invasiveness. The differential effects of these antiestrogenic agents on invasion correlated with changes in production of collagenase IV, while no significant change was seen in the chemotactic activity of the cells. Invasiveness was increased by 17 beta-estradiol or 4-hydroxytamoxifen after a few hours of treatment and was rapidly lost when 17 beta-estradiol was withdrawn. Stimulation of invasiveness with 17 beta-estradiol was blocked by the antiestrogen, LY117018. Cells from the MDA-MB-231 line which lacks estrogen receptors were not affected by estrogen or antiestrogen in terms of proliferation or invasion. These studies indicate that the invasiveness of MCF-7 cells is regulated by antiestrogens through the estrogen receptor and may be mediated by collagenase IV activity. Antiestrogens which reduce both the proliferation and invasiveness of these cells may be interesting new candidates for clinical application. 相似文献
4.
Colin K. W. Watts Kimberley J. E. Sweeney Andrea Warlters Elizabeth A. Musgrove Robert L. Sutherland 《Breast cancer research and treatment》1994,31(1):95-105
The molecular mechanisms by which antiestrogens inhibit breast cancer cell proliferation are not well understood. Using cultured breast cancer cell lines, we studied the effects of antiestrogens on proliferation and cell cycle progression and used this information to select candidate cell cycle regulatory genes that are potential targets for antiestrogens. Under estrogen- and serum-free conditions antiestrogens inhibited proliferation of MCF-7 cells stimulated with insulin. Cells were blocked at a point in G1 phase. These effects are comparable with those in serum- and estrogen-containing medium and were also seen to a lesser degree in nude mice bearing MCF-7 tumors. Similar observations with other peptide mitogens suggest that the process inhibited by antiestrogens is common to estrogen and growth factor activated pathways. Other studies have identified G1 cyclins as potential targets for growth factor and steroid hormone/steroid antagonist regulation of breast epithelial cell proliferation. In MCF-7 cells growing in the presence of fetal calf serum, cyclin D1 mRNA was rapidly down-regulated by steroidal and nonsteroidal antiestrogens by an apparently estrogen receptor mediated mechanism. Cyclin D1 gene expression was maximally inhibited before effects on entry into S phase and inhibition was therefore not merely a consequence of changes in cell cycle progression. Together with data on the effects of antiestrogens in serum-free conditions [1], these results suggest down-regulation of cyclin D1 by antiestrogens may be a general phenomenon in estrogen receptor-positive breast cancer cells, independent of culture conditions and class of antiestrogen. These observations are compatible with the hypothesis that reductions in cyclin D1 levels may mediate in part the action of antiestrogens in blocking entry of cells into S phase. 相似文献
5.
目的观察靶向survivin的siRNA联合三苯氧胺对乳腺癌MCF-7细胞增殖和凋亡的影响.方法构建靶向survivin的siRNA真核表达载体,采用lipofec-tamineTM2000转染乳腺癌MCF-7细胞,采用MTT法检测靶向survivin的siRNA联合三苯氧胺对MCF-7细胞增殖的影响;采用流式细胞术检测靶向survivin的siRNA联合三苯氧胺诱导MCF-7细胞凋亡的作用.结果靶向survivin的siRNA和三苯氧胺均可抑制MCF-7细胞增殖并诱导其一定程度的凋亡;当联合应用靶向survivin的siRNA和三苯氧胺时,上述作用得到显著加强(P<0.05).结论利用RNA干扰技术阻断survivin基因的表达可以显著增强MCF-7细胞对三苯氧胺的敏感性,靶向survivin的RNA干扰技术在乳腺癌的内分泌治疗中具有重要的潜在价值. 相似文献
6.
目的:观察靶向survivin的siRNA联合三苯氧胺对乳腺癌MCF-7细胞增殖和凋亡的影响.方法:构建靶向survivin的siRNA真核表达载体,采用lipofec-tamineTM2000转染乳腺癌MCF-7细胞,采用MTT法检测靶向survivin的siRNA联合三苯氧胺对MCF-7细胞增殖的影响;采用流式细胞术检测靶向survivin的siRNA联合三苯氧胺诱导MCF-7细胞凋亡的作用.结果:靶向survivin的siRNA和三苯氧胺均可抑制MCF-7细胞增殖并诱导其一定程度的凋亡;当联合应用靶向survivin的siRNA和三苯氧胺时,上述作用得到显著加强(P<0.05).结论:利用RNA干扰技术阻断survivin基因的表达可以显著增强MCF-7细胞对三苯氧胺的敏感性,靶向survivin的RNA干扰技术在乳腺癌的内分泌治疗中具有重要的潜在价值. 相似文献
7.
目的 探讨在乳腺癌细胞系MCF-7中下调S期激酶相关蛋白(Skp2)表达诱导细胞凋亡及其机制。方法 应用RNAi方法在体外下调乳腺癌细胞系MCF-7中Skp2的表达水平,48小时后表阿霉素处理细胞,TUNEL和Hoechst 33258染色检测凋亡,Western Blot检测细胞周期调控相关因子及凋亡相关蛋白表达情况,研究其机制。结果 下调MCF-7中Skp2表达水平后,乳腺癌细胞凋亡增加。下调Skp2使p27、p21和CyclinE蛋白表达水平升高。表阿霉素处理MCF-7细胞后,Skp2蛋白水平下调。Skp2 siRNA与表阿霉素有协同诱导凋亡的作用,p53蛋白水平升高。结论 p27、p21和CyclinE在通过下调Skp2诱导的凋亡中发挥作用。Skp2 siRNA和表阿霉素协同诱导细胞凋亡,与p53依赖的凋亡途径有关。Skp2可能是乳腺癌治疗的靶点。 相似文献
8.
靶向survivin的siRNA抑制乳腺癌MCF-7细胞增殖并诱导其凋亡 总被引:11,自引:2,他引:11
目的 观察靶向survivin的siRNA对乳腺癌细胞增殖和凋亡的影响。方法构建靶向survivin的siRNA真核表达载体,采用Lipofectamine^TM2000转染乳腺癌细胞MCF-7,采用半定量RTPCR、免疫组化技术检测转染前后MCF-7细胞survivin基因表达的变化;采用MTT法检测对MCF-7细胞增殖的抑制作用;采用TUNEL法检测诱导MCF-7细胞凋亡的作用。结果靶向survivin的序列特异性siRNA可以高效抑制MCF-7细胞survivin基因的表达,在mRNA水平其表达抑制率为64.9%;在蛋白质水平其表达抑制率为79.7%;转染靶向survivin的siRNA真核表达载体可以显著抑制MCF-7细胞的增殖,细胞重新接种24、48h后,其增殖抑制率分别为31.6%和33.0%;转染后24h可以诱导12.9%的细胞凋亡。结论利用RNA干扰技术阻断survivin基因的表达可以显著抑制MCF-7细胞的增殖,并在一定程度上诱导其自发凋亡,靶向survivin的RNA干扰技术在乳腺癌的基因治疗中具有一定的价值。 相似文献
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10.
Apogossypolone抑制乳腺癌MCF-7细胞增殖及诱导细胞凋亡 总被引:3,自引:0,他引:3
目的:研究Apogossypolone(ApoG2)对人乳腺癌细胞系MCF-7体外的增殖抑制和诱导凋亡作用.方法:采用MTT法和平板克隆形成实验检测ApoG2作用于MCF-7细胞后,对其体外增殖和细胞克隆形成能力的影响;Hoechst 33258荧光染色法观察ApoG2(20μmol/L)作用48 h后,细胞凋亡的形态学变化;FCM检测ApoG2作用于MCF-7细胞后的细胞凋亡率以及细胞周期的变化.结果:ApoG2药物在浓度为5μmol/L、作用14 d后就能对MCF-7细胞增殖起到明显的抑制作用,且呈剂量和时间依赖性;平板克隆实验表明,经不同浓度(5、10和20 μmol/L)ApoG2作用14 d后,对MCF-7细胞克隆形成能力呈现明显的抑制作用,且同样呈剂量和时间依赖性.Hoechst 33258荧光染色结果显示,ApoG2与MCF-7细胞共培养72 h后,细胞呈现明显的核固缩和碎裂、染色质凝集和凋亡小体形成等细胞凋亡现象.FCM检测结果显示,20 μmol/LApoG2作用72 h后,细胞凋亡率较对照组升高,而且S期和G2/M期细胞比例较对照组升高(P<0.05).结论:ApoG2能够抑制人乳腺癌细胞系MCF-7的增殖和克隆形成,阻滞细胞周期,诱导细胞凋亡. 相似文献
11.
《Annals of oncology》2010,21(2):263-268
BackgroundIn order to study the anticancer effects and cellular apoptosis pathways induced by daidzein.Materials and methodsWe used the human MCF-7 breast cancer cell line as a model and examined the apoptosis by Hoechst–propidium iodide staining fluorescence imaging and flow cytometry.ResultsOur data indicated that daidzein induces antiproliferative effects in a concentration- and time-dependent manner. We demonstrated that daidzein-induced apoptosis in MCF-7 cells was initiated by the generation of reactive oxygen species (ROS). Furthermore, we showed that this daidzein-induced ROS generation was accompanied by disruption of mitochondrial transmembrane potential, down-regulation of bcl-2, and up-regulation of bax, which led to the release of cytochrome C from the mitochondria into the cytosol, which, in turn, resulted in the activation of caspase-9 and caspase-7, and ultimately in cell death. The induction of the mitochondrial caspase-dependent pathway was confirmed by pretreatment with pan-caspase inhibitor z-VAD-fmk and antioxidant N-acetyl-L-cysteine.ConclusionAccordingly, daidzein could induce breast cancer cell apoptosis through the mitochondrial caspase-dependent cell death pathway. 相似文献
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13.
存活素siRNA表达质粒的构建及其对MCF-7细胞细胞周期和增殖的调控 总被引:13,自引:2,他引:13
背景与目的:存活素特异地在肿瘤组织中过表达,许多报道表明抑制它的功能对肿瘤治疗有利。RNA干扰被证明是一项能有效而特异地抑制基因表达的新技术。本研究的目的在于构建存活素基因的小分子干扰RNA(siRNA)表达质粒,并检测该质粒在乳腺癌细胞中的生物学功能。方法:应用含有u6启动子的mu6pro载体构建存活素siRNA质粒,RT-PCR和Western blot法检测该质粒转染MCF-7细胞后存活素表达的变化,流式细胞仪和MTT法分别检测其对细胞周期和对细胞增殖的影响。结果:存活素siRNA质粒明显下调MCF-7细胞中存活素的表达,阻断细胞周期在G1期,显著抑制细胞增殖,促进细胞凋亡。结论:通过RNA干扰,存活素的表达在mRNA和蛋白质水平上被下调。 相似文献
14.
目的:研究抑制促分裂原活化蛋白激酶/细胞外信号调节激酶-激酶3(mitogen—activated protein/extraeellular signal regulated kinase—kinase3,MEKK3)基因表达促进TRAIL诱导乳腺癌MCF-7细胞凋亡的作用,寻找乳腺癌临床治疗新策略。方法:应用MTT法检测人TRAIL对MCF-7细胞生长的抑制作用。合成MEKK3-siRNA,应用脂质体介导MEKK3-siRNA转染人人乳腺癌细胞MCF-7,以RT,PCR和Westernblotting法检测MCF-7细胞MEKK3mRNA和蛋白的表达。应用MTr法和流式细胞术检测MEKK3-siRNA与TRAIL联合处理后MCF-7细胞的增殖和凋亡。结果:TRAIL具有抑制MCF-7细胞增殖作用,但其抑制作用较弱。MEKK3-siRNA转染后能有效而稳定地抑制MCF-7细胞中MEKK3mRNA和蛋白的表达(P〈0.01)。TRAIL与MEKK3-siRNA联合处理MCF-7细胞较TRAIL单独处理更明显地抑制细胞增殖活力(P〈0.05),更明显地增加细胞凋亡率(P〈0.01)。结论:siRNA沉默MEKK3基因能显著促进TRAIL对乳腺癌MCF-7细胞凋亡的诱导作用,为探讨乳腺癌治疗新方案提供了实验依据。 相似文献
15.
Acquired antiestrogen resistance in MCF-7 human breast cancer sublines is not accomplished by altered expression of receptors in the ErbB-family 总被引:3,自引:0,他引:3
Larsen SS Egeblad M Jäättelä M Lykkesfeldt AE 《Breast cancer research and treatment》1999,58(1):41-56
Development of acquired resistance against antiestrogen treatment is a serious problem in human breast cancer, and knowledge of alterations resulting in resistance is important for selection of further treatment. To mimic the clinical situation we have established a series of MCF-7 human breast cancer cell lines by long term treatment with the antiestrogens tamoxifen, ICI 164,384, and ICI 182,780. Common for these cell lines is a decreased expression of the estrogen receptor (ER). In human breast cancer, lack of response to endocrine therapy is often associated with decreased expression of the estrogen receptor and increased expression of epidermal growth factor receptor (EGFR) and/or HER-2/neu (ErbB-2). Our antiestrogen resistant cell lines did not express altered levels of EGFR, HER-2/neu, ErbB-3, or ErbB-4. Estrogen and antiestrogen regulation of HER-2/neu expression was essentially similar in parent and resistant MCF-7 cells. Treatment with antibodies to HER-2/neu (HerceptinTM) did not affect growth of MCF-7 cells or resistant cells, indicating that in this in vitro model system, acquired antiestrogen resistance does not emerge from activation of the HER-2/neu signaling pathway. In MCF-7 cells transfected with HER-2/neu and/or ErbB-3, overexpression alone did not result in resistance. However, addition of heregulinl-1 abolished the inhibitory activity of ICI 182,780 on both vector and HER-2/neu/ErbB-3 transfected MCF-7 cells, demonstrating that activation of the HER-2/neu receptor signaling pathway can override the growth inhibitory effect of ICI 182,780. 相似文献
16.
阿蒂莫耶番荔枝内酯诱导MCF—7/ADR细胞凋亡的研究 总被引:1,自引:1,他引:1
目的:探讨阿蒂莫耶番荔枝内酯对有多药抗药性的阿霉素的人乳腺癌细胞MCF-7/ADR凋亡的诱导作用,揭示其抗癌机制。方法;用MTT法观察药物对细胞的生长抑制作用,TUNEL法,流式细胞仪及光镜,电镜等技术研究癌细胞凋亡。结果;药物对癌细胞有明显生长抑制作用,IC50O 10.5mg/L;光1镜,电镜可见染色质浓缩,聚集核膜下,核碎裂等改变;流式细胞仪检测到凋亡峰;10mg/L药物作用48小时,TUN 相似文献
17.
Growth of the human breast cancer cell line MCF-7 is known to be inhibited both by antiestrogens such as 4-hydroxytamoxifen (OHTAM) and by retinoic acid (RA). Uncloned MCF-7 cells (UNC) and two cloned sublines, one sensitive to antiestrogens (E-3) and the other resistant to them (RR), were used in this study. Growth of UNC and E-3 was inhibited by either OHTAM (10(-7) M) or RA (10(-6) M), and this inhibition could not be overcome by the simultaneous addition of estradiol. Subline RR, which was originally selected for resistance to tamoxifen, was resistant to both OHTAM and RA as measured by either growth in culture or colony forming ability. RR was resistant to RA at all concentrations tested between 10(-9) M and 10(-6) M. The inhibition of uncloned MCF-7 cells by RA was dose dependent between 10(-9) M and 10(-6) M. Subline E-3, however, exhibited a mixed response to RA. At 10(-9) M and 10(-8) M, growth was stimulated, but at 10(-7) M and 10(-6) M it was inhibited. The level of estrogen receptor was measured in the same experiment by using a whole cell assay. In the uncloned MCF-7 cultures and in both the RR and E-3 sublines the level of estrogen receptor was increased between 50 and 200% by RA. The production of plasminogen activator by MCF-7 cells is stimulated by estrogen. RA had a dual effect on plasminogen activator production. In the absence of estrogen, RA inhibited production below the unstimulated level, but in cells stimulated by estrogen, RA increased plasminogen activator production. The results reported here support possible interactions between the mechanisms by which cells respond to estrogen, antiestrogens, and retinoids. 相似文献
18.
Sunyoung Park Sun Young Yoon Kyung-Eun Kim Ha Reum Lee Dae Young Hur Hyunkeun Song Daejin Kim Sa Ik Bang Dae-Ho Cho 《Cancer letters》2009
Interleukin-18 (IL-18) has recently been shown to have a pro-cancer effect in various cancers. Increased serum levels of both IL-18 and transferrin in cancer patients are correlated with its malignancy. However, the relationship between transferrin and IL-18 is not well understood. Here, we show that exogenous transferrin enhanced breast cancer cell proliferation and the proliferation rate was reduced when endogenous transferrin expression was inhibited by transferrin siRNA. The expression of endogenous transferrin was found to be regulated by IL-18. Furthermore, we found that the MAPK pathway is involved in IL-18-induced transferrin production. In conclusion, IL-18 is suggested as an inducer of endogenous transferrin expression in breast cancer cells. 相似文献
19.
目的 观察不同浓度瘦素对人类乳腺癌细胞MCF-7增殖及凋亡的影响,探讨瘦素在乳腺癌发生及发展中的作用。方法 采用四甲基偶氮唑蓝(MTT)比色法检测不同浓度瘦素对体外培养的MCF-7细胞生长的增殖作用,流式细胞术分析细胞周期的分布, Annexin V-FITC/PI染色法检测细胞凋亡。结果 MTT比色法结果显示:不同浓度瘦素作用24、48、72 h能够促进MCF-7细胞增殖,浓度与时间不存在交互作用(F=0.919,P=0.523),浓度及时间各自的主效应均差异有统计学意义(F=12.699,P=0.000; F=647.881,P=0.000)。多重比较结果显示:200 ng/ml及400 ng/ml瘦素组与对照组相比差异有统计学意义(P=0.007;P=0.000),不同时间段之间均差异有统计学意义(P=0.000)。流式细胞术分析显示100、400 ng/ml瘦素作用48 h后,G0/G1期细胞比例下降14.42 %(F=10.464,P=0.044),S期比例分别上升7.57 %、22.19 %(F=47.361,P=0.005),G2/M期变化差异无统计学意义(F=1.77,P=0.311)。未发现瘦素对MCF-7细胞早期凋亡的抑制作用。结论 瘦素能够促进乳腺癌细胞MCF-7的增殖并改变生长周期,但不会抑制凋亡,提示瘦素可能在乳腺癌的发展中发挥促进作用。 相似文献
20.
Down-regulation of P-glycoprotein expression in MDR breast cancer cell MCF-7/ADR by honokiol 总被引:8,自引:0,他引:8
P-glycoprotein accounts for the most intrinsic and acquired cancer multidrug resistance. To inhibit the expression of P-glycoprotein is one of the effective ways to reverse cancer drug resistance. Honokiol, a naturally occurring compound, has been demonstrated to combat cancer through mechanisms including inhibition of angiogenesis and induction of apoptosis. Here, we show that honokiol down-regulated the expression of P-glycoprotein at mRNA and protein levels in MCF-7/ADR, a human breast MDR cancer cell line. The down-regulation of P-glycoprotein was accompanied with a partial recovery of the intracellular drug accumulation, and of the sensitivities toward adriamycin. This study reveals a novel function of honokiol as an anti-cancer agent. 相似文献