三氧化二砷诱导人横纹肌肉瘤RD细胞株凋亡的实验研究

[目的]研究三氧化二砷(As2O3)体外抑制人胚胎型横纹肌肉瘤细胞株RD(RD细胞株)增殖及诱导凋亡的作用. [方法]以不同浓度的As2O3处理RD细胞株,用四甲基偶氮唑蓝(MTT)法测定细胞增殖情况,流式细胞仪检测细胞的DNA含量及凋亡率. [结果]4～8 μmol/L hs2O3能以时间剂量依赖的方式抑制RD细胞株的增殖,并诱导其凋亡,在流式细胞仪DNA线条图上出现典型的亚二倍体“凋亡峰”,细胞周期阻滞于G0/G1期,S期和G2/M期细胞减少;低浓度As2O3(＜2μmol/L)则有轻微促增殖作用,细胞周期向S期和G2/M期集中,G0/G1期细胞减少. [结论]As2O3浓度≥4μmol/L时,可明显抑制RD细胞株的生长并促进细胞凋亡,其临床应用还需进一步研究.     

甲基硒酸对乳腺癌细胞体外凋亡和细胞周期的影响

目的:观察甲基硒酸(MSA)对乳腺癌细胞系MCF-7凋亡和细胞周期的影响。方法:以不同浓度的MSA作用乳腺癌细胞系MCF-7,在不同时间内,观察不同浓度MSA对乳腺癌细胞凋亡率和细胞周期的影响。流式细胞术检测细胞凋亡率和细胞周期。结果:①MSA可诱导乳腺癌细胞体外凋亡,随着药物浓度及作用时间的增加,乳腺癌细胞凋亡率逐渐增加。②MSA可改变乳腺癌的细胞周期,将乳腺癌细胞周期阻断在G1期,并且减少S期细胞的比例。结论:MSA可诱导乳腺癌细胞体外凋亡、调控乳腺癌细胞周期。MSA有望成为预防和治疗乳腺癌的一种新方法。     

2-甲氧雌二醇诱导人子宫内膜癌细胞株凋亡的实验研究

目的:探讨2-甲氧雌二醇(2-ME)对人子宫内膜癌细胞株KLE细胞凋亡的影响。方法:选用人子宫内膜癌细胞株KLE进行体外培养,实验组加入不同浓度2-ME,对照组不含2-ME。用电子显微镜(电镜)观察细胞形态变化;流式细胞仪(FCM)观察细胞的凋亡率及细胞周期的变化,以及免疫组织化学方法检测P53和Bcl-2的表达强度。结果:2-ME作用后G0/G1期细胞增加,并伴随G0/G1期细胞的增加,出现细胞凋亡峰和凋亡率的升高(P<0.05)。电镜下观察到KLE细胞染色体边集、核固缩、凋亡小体,P53和bc1-2阳性表达率明显下降。结论:2-ME对人子宫内膜癌KLE细胞株有诱导凋亡作用。     

三氧化二砷联合化疗诱导肝癌HepG2细胞凋亡作用机制研究

目的探讨三氧化二砷（As2O3）联合化疗诱导肝癌HepG2细胞凋亡的作用机制。方法体外培养人肝癌HepG2细胞株，分别给予As2O3和顺铂单独或联合处理，用细胞免疫组化酶法检测Bax、Bcl-2、Fas蛋白的表达情况；采用流式细胞仪观察人肝癌HepG2细胞株的细胞周期变化，细胞DNA含量的分布，测定不同作用时间后端粒酶活性的变化情况。结果As2O3，组肝癌HepG2细胞的Bax和Fas基因的表达明显增加，Bcl-2基因的表达明显降低；在DNA直方图上可见在G1期细胞前凋亡细胞呈现典型特征性的亚二倍体凋亡峰，使S期细胞减少，将细胞生长周期阻滞于G2/M期，联合用药组比单独用药组作用明显增强。结论As2O3及联合化疗能诱导肝癌细胞凋亡，其主要机制与增强Bax、Fas的表达，下调Bcl-2的表达，将细胞生长周期阻滞于G2/M期，阻止细胞的有丝分裂，抑制端粒酶活性有关。     

家蝇幼虫抗菌蛋白诱导黑色素瘤A375细胞凋亡的探讨

目的 观察家蝇幼虫抗菌蛋白对黑色素瘤A375细胞凋亡的诱导，探讨其抗肿瘤作用机制。方法 流式细胞仪测定家蝇幼虫抗菌蛋白对细胞周期和细胞凋亡的影响；HE染色法观察细胞凋亡形态，并测定细胞凋亡指数（AI）。结果 在0．10％组A375的G／M期升高，S期降低。0．50％组G0／G1期、G2／M期和AI／PI（增殖指数）显著增加，S期及PI明显减少。2．50％组S期、PI、AI／PI及细胞凋亡率（Ap）显著增高，G0／G1期大大减少。12．50％组G2／M期、PI、AI／PI及Ap显著增高，G0／G1期和S期明显下降。结论 家蝇幼虫抗菌蛋白对黑色素瘤A375细胞有明显的抑杀作用；家蝇幼虫抗菌物质可能系通过细胞周期阻滞以及诱导细胞凋亡而抑制黑色素瘤A375细胞生长。     

染料木质素对人乳腺癌细胞BCaP-37凋亡的诱导作用

采用荧光显微镜观察了染料木质素诱导体外培养的人体乳腺髓样癌细胞株BCaP 37凋亡的形态学特征 ,并用流式细胞仪检测了染料木质素对乳腺癌细胞株细胞周期、凋亡率和bcl 2基因表达的影响。结果显示经丫啶橙染色后 ,荧光显微镜下可见到乳腺癌细胞出现细胞凋亡的特征性形态学改变 ;染料木质素能阻断细胞由G1 期向S期移行 ,使G0 -G1 期细胞数目明显增多 ,并能诱导细胞发生凋亡 ,呈现剂量 效应关系 ;bcl 2基因表达量与染料木质素浓度呈负相关。故染料木质素可通过抑制bcl 2蛋白表达诱导细胞凋亡     

高温联合化疗药对人乳腺癌细胞杀伤作用研究

目的:探讨高温联合化疗药物诱导人乳腺癌细胞株(MDA-MB-435S)凋亡和细胞周期各时相变化的分子机理。方法:应用43℃水浴箱高温诱导MDA-MB-435S细胞株,并联合使用5-Fu进行细胞杀伤,通过MTT比色法和流式细胞仪检测细胞凋亡率和细胞周期各时相的变化。结果;高温组、高温联合5-Fu组细胞凋亡率及细胞周期时相变化与对照组差异显著(P<0.01)。结论:高温联合5-Fu可提高乳腺癌细胞凋亡率、可抑制乳腺细胞周期G1期向S期的进程,S期细胞减少、G2M期细胞相对增高。     

ω-3多不饱和脂肪酸对人胃癌BGC-823细胞周期及凋亡的影响

目的 利用人胃癌BGC-823细胞体外培养实验，观察不同浓度ω-3多不饱和脂肪酸（EPA、DHA）对肿瘤细胞周期及凋亡的影响。方法 采用MTT法、凋亡形态学检查和流式细胞检测肿瘤细胞周期及凋亡情况。结果 30和45μg／ml的EPA或DHA可显著降低肿瘤细胞存活率（P〈0．001）；Hoechst33258染色和透射电镜可见细胞凋亡现象；流式细胞检测bcl-2基因蛋白表达明显下降（P〈0．05），G0／G1细胞明显增加，G2／M和S期细胞明显减少（P〈0．05）。结论 ω-3多不饱和脂肪酸可诱导人胃癌BGC-823细胞凋亡和阻滞细胞在G0／G1期，细胞增殖受到抑制。     

三氧化二砷联合化疗诱导肝癌HepG2细胞凋亡作用机制研究

目的探讨三氧化二砷(As2O3)联合化疗诱导肝癌HepG2细胞凋亡的作用机制。方法体外培养人肝癌HepG2细胞株,分别给予As2O3和顺铂单独或联合处理,用细胞免疫组化酶法检测Bax、Bcl-2、Fas蛋白的表达情况;采用流式细胞仪观察人肝癌HepG2细胞株的细胞周期变化,细胞DNA含量的分布,测定不同作用时间后端粒酶活性的变化情况。结果As2O3组肝癌HepG2细胞的Bax和Fas基因的表达明显增加,Bcl-2基因的表达明显降低;在DNA直方图上可见在G1期细胞前凋亡细胞呈现典型特征性的亚二倍体凋亡峰,使S期细胞减少,将细胞生长周期阻滞于G2/M期,联合用药组比单独用药组作用明显增强。结论As2O3及联合化疗能诱导肝癌细胞凋亡,其主要机制与增强Bax、Fas的表达,下调Bcl-2的表达,将细胞生长周期阻滞于G2/M期,阻止细胞的有丝分裂,抑制端粒酶活性有关。     

TAM和MPA联合用药对卵巢癌顺铂耐药细胞株SKOV-3/DDP生长的影响

目的:探讨三苯氧胺(tamoxifen,TAM)和甲孕酮(medroxyprogesterone,MPA)联合用药对顺铂耐药的卵巢癌细胞株SKOV-3／DDP生长影响的机制。方法:用四甲基偶氮唑蓝(MTT)法、电子显微镜及流式细胞仪测定用药前后SKOV-3／DDP细胞生长、形态学、周期及凋亡率的变化。结果:低浓度的TAM和MPA联合用药组对SKOV-3／DDP细胞生长无影响。高浓度用药组可抑制卵巢癌细胞生长,电子显微镜可观察到细胞凋亡的形态学变化,1×10-6~1×10-4mol／L的TAM和MPA联合用药组在抑制细胞增殖的同时尚可诱导细胞凋亡,其细胞凋亡率分别为4.06％、11.77％、19.3％,两两比较,差异均有显著性(P<0.05)。结论:TAM和MPA联合用药既对顺铂耐药的卵巢癌细胞有杀灭作用,又因其较小的副作用及肯定的临床意义有可能成为卵巢癌有效的化疗辅助用药。     

FTY720对人乳腺癌MCF-7细胞、喉癌Hep-2细胞增殖的影响

目的:探讨FTY720抑制人乳腺癌MCF-7细胞和喉癌Hep-2细胞增殖作用。方法:将人乳腺癌MCF-7细胞和喉癌Hep-2细胞用不同浓度FTY720作用48 h,MTT法检测细胞活性,流式细胞术检测其对细胞周期及凋亡率的影响。结果:MTT结果显示,FTY720对乳腺癌MCF-7细胞和喉癌Hep-2细胞增殖均有抑制作用,而且对喉癌Hep-2细胞抑制作用的浓度依赖性高于对乳腺癌细胞的作用;流式细胞术检测结果显示,FTY720可将乳腺癌MCF-7细胞阻滞于G1期,将喉癌Hep-2细胞阻滞于G2/M期,并明显促进乳腺癌MCF-7细胞凋亡。结论:一定浓度的FTY720能明显抑制体外培养的乳腺癌和喉癌细胞增殖,调节其细胞周期,诱导乳腺癌MCF-7细胞凋亡。     

Effects of decabrominated diphenyl ether (PBDE-209) in regulation of growth and apoptosis of breast, ovarian, and cervical cancer cells

Background: Polybrominated diphenyl ethers (PBDEs), commonly used in building materials, electronics, plastics, polyurethane foams, and textiles, are health hazards found in the environment.Objective: In this study we investigated the effects of PBDE-209, a deca-PBDE, on the regulation of growth and apoptosis of breast, ovarian, and cervical cancer cells as well as the underlying protein alterations.Methods: We used MCF-7 and MCF-7/ADR (multidrug-resistant MCF-7) breast cancer cell lines, the HeLa cervical cancer cell line, the OVCAR-3 ovarian cancer cell line, and the normal CHO (Chinese hamster ovary) cell line to assess the effects of PBDE-209 using cell viability, immunofluorescence, and flow cytometric assays. Western blot assays were used to detect changes in protein expression. To assess the effects of PBDE-209 on apoptosis, we used the protein kinase Cα (PKCα) inhibitor Gö 6976, the extracellular signal-regulated kinase (ERK) inhibitor PD98059, and tamoxifen.Results: Our data indicate that PBDE-209 increased viability and proliferation of the tumor cell lines and in CHO cells in a dose- and time-dependent manner. PBDE-209 also altered cell cycle distribution by inducing the S phase or G₂/M phase. Furthermore, PBDE-209 partially suppressed tamoxifen-induced cell apoptosis in the breast cancer cell lines (MCF-7 and MCF-7/ADR) but suppressed Gö 6976- and PD98059-induced apoptosis in all cell lines. At the molecular level, PBDE-209 enhanced PKCα and ERK1/2 phosphorylation in the cell lines.Conclusions: Our data demonstrate that PBDE-209 is able to promote proliferation of various cancer cells from the female reproductive system and normal ovarian CHO cells. Furthermore, it reduced tamoxifen, PKCα, and ERK inhibition-induced apoptosis. Finally, PBDE-209 up-regulated phosphorylation of PKCα and ERK1/2 proteins in tumor cells and in CHO cells.     

岩大戟内酯B诱导乳腺癌MCF-7细胞凋亡的实验研究

目的：探讨岩大戟内酯B 对MCF-7 乳腺癌细胞生长抑制作用及其机理.方法：通过MTT 法观察各组细胞增殖能力;采用流式检测技术分析细胞周期;利用Annexin V/PI 双染检测细胞凋亡情况;电镜观察细胞形态.结果：随着岩大戟内酯B 浓度增加对MCF-7 的抑瘤率增高;G2/M 期细胞减少,S 期细胞增多,出现S 期阻滞;晚期凋亡和死亡的细胞增加;电镜显示加药后细胞出现核固缩,细胞器空泡变性,凋亡小体形成,甚至出现坏死.结论：岩大戟内酯B 对乳腺癌MCF-7 细胞生长明显抑制作用,并诱导肿瘤细胞凋亡,为临床开发新药提供理论依据.     

Anticancer activities of an anthocyanin-rich extract from black rice against breast cancer cells in vitro and in vivo

Anthocyanins widely present in human diet and have a variety of health effects. This study investigates the anticancer effects of an anthocyanin-rich extract from black rice (AEBR) on breast cancer cells in vitro and in vivo. AEBR reduced the viability of breast cancer cell lines MCF-7 (ER(+), HER2/neu(-)), MDA-MB-231 (ER(-), HER2/neu(-)), and MDA-MB-453 (ER(-), HER2/neu(+)) and induced apoptosis in MDA-MB-453 cells via the intrinsic pathway in vitro by activating caspase cascade, cleaving poly (ADP-ribose) polymerase (PARP), depolarizing mitochondrial membrane potential, and releasing cytochrome C. Oral administration of AEBR (100 mg/kg/day) to BALB/c nude mice bearing MDA-MB-453 cell xenografts significantly suppressed tumor growth and angiogenesis by suppressing the expression of angiogenesis factors MMP-9, MMP-2, and uPA in tumor tissue. Altogether, this study suggests the anticancer effects of AEBR against human breast cancer cells in vitro and in vivo by inducing apoptosis and suppressing angiogenesis.     

内分泌及化疗对乳腺肿瘤干细胞的实验研究

目的探讨研究内分泌及化疗两种治疗方式对ER＋乳腺癌患者中乳腺癌CD44＋CD24low细胞亚群的影响．以期推动ER‘乳腺癌的临床治疗。方法取ER’乳腺癌患者中乳腺癌细胞株MCF-7,进行体外培养,无水乙醇、雌二醇、以及雌二醇联合三苯氧胺的培养观察。随后选取雌二醇组中处于对数生长期的细胞,分别进行紫杉醇、吉西他滨以及阿霉素的培养观察。统计分析各组的分析结果。结果雌二醇培养组与联合培养组,细胞培养7d、10d以及15d后,细胞中CD44＋CD24-aow表现细胞所占的比例相比较或其分别与对照组比较均具有显著性差异（P〈O．05）。而紫杉醇、吉西他滨以及阿霉素培养组中,细胞中CD44＋CD24／Low表现细胞比例在培养1d后与对照组比较有显著性差异（P〈0．05）;在培养15d后与对照组比较无显著性差异（P〉0．05）。结论内分泌以及化疗对Elk＋乳腺癌细胞均有重大影响,对于ER＋乳腺癌根治具有重要意义,值得进一步研究探讨。     

维生素C和紫杉醇联合作用对乳腺癌细胞MCF-7增殖、细胞周期和凋亡的影响

目的研究维生素C(VC)与紫杉醇联合作用,对人乳癌MCF-7细胞增殖、细胞周期及凋亡的影响。方法 MTT法检测细胞增殖,激光共聚焦观察自噬,流式细胞术测定细胞周期分布和凋亡率变化。结果 VC单独作用,可促进MCF-7的增殖;紫杉醇单独作用及与VC联合作用均可抑制MCF-7的生长,并将细胞阻滞在G2期及诱导细胞凋亡;紫杉醇与VC联合作用对细胞增殖的影响,与二者的用药顺序有关,紫杉醇作用24后联合应用VC,可协同抑制MCF-7的增殖,并且,紫杉醇的IC50最低,即MCF-7对紫杉醇的敏感性最高。结论紫杉醇与VC联合作用对人乳腺癌MCF-7细胞增殖的影响,与二者的用药顺序有关,其作用的机制与影响细胞周期分布和诱导细胞凋亡有关。     

Growth inhibition and induction of apoptosis in MCF-7 breast cancer cells by fermented soy milk

The effect of a fermented soy milk product (FSP) on various human breast carcinoma cell lines was investigated, and it was shown to have a growth-inhibitory effect, especially on MCF-7 cells. Thus the MCF-7 cell line was used to study the mechanism of action. In female severe combined immune deficiency mice implanted with MCF-7 cells, pretreatment with FSP significantly inhibited tumor growth. The inhibitory effect of FSP on MCF-7 cells seemed to be caused by the additive effects of a wide variety of constituents. The active components of FSP are mainly in the water phase, and the lipid-soluble fraction, which includes the soy isoflavones such as genistein and daidzein, is relatively ineffective. A variety of methods were used to demonstrate that FSP caused apoptotic cell death in MCF-7 cells. FSP induced generation of reactive oxygen species (ROS). Growth inhibition and ROS generation induced by FSP could be inhibited by catalase and deferoxamine, indicating that the ROS production probably was the cause of this apoptotic cell death. This study suggests that FSP retards tumor growth in vivo and can trigger apoptosis in vitro. It may, therefore, be a potential nutritional supplement in chemotherapy.     

金雀异黄素的抗雌激素效应

应用雌激素耗尽的方法诱导乳腺癌细胞株MCF - 7和T47D发生凋亡 ,从而探讨金雀异黄素(genistein ,GS)对凋亡的影响作用。结果表明 ,同抗雌激素药物它莫西芬类似 ,3 2× 10 - 6 mol L及 96× 10 - 6 mol LGS可加剧雌激素耗尽所诱导的细胞凋亡作用 ,且这种效应存在着良好的剂量 -效应关系。结果提示GS对女性乳腺癌发病率的预防作用是通过其抗雌激素效应诱导细胞的凋亡而实现的     

Novel interactions of vitamin E and estrogen in breast cancer

The prevention of breast cancer through dietary modification is an active area of clinical and epidemiological research. It has been proposed that dietary supplementation of vitamin E may reduce a woman's risk of developing breast cancer. However, the exact mechanism remains unknown. alpha-Tocopherol is the most biologically active form of vitamin E. We investigated the effect of vitamin E (alpha-tocopherol) on breast cancer cell growth. A dose-dependent inhibition of cell proliferation was found in estrogen receptor (ER)-positive cells showing a potent suppression of growth at 100 microM vitamin E in MCF-7 (53%) and T47D (75%) cells. Vitamin E reduced significantly the response of both cell lines to estrogen (10 nM), and cell proliferation was decreased in MCF-7 and T47D cells by 69% and 84%, respectively. No growth inhibition was observed when cells were grown in the absence of estrogen. Vitamin E altered and decreased the growth inhibition induced by tamoxifen (10 microM) in MCF-7 (33%) and T47D (54%) cells. In addition, the immunostaining of ER of MCF-7 cells was reduced by 30% in the presence of vitamin E, suggesting an effect of vitamin E on the expression of ER. This provides evidence that vitamin E may inhibit ER-positive cell growth by altering the cellular response to estrogen.