甘草酸对人乳腺癌细胞增殖抑制和诱导凋亡作用的研究

目的　探讨甘草酸对人乳腺癌细胞 (MCF 7)增殖抑制和诱导凋亡的作用 ,并探讨凋亡发生与细胞内Ca2 浓度( [Ca2 ]i)变化的关系。方法　 2 .5～ 12 .5mmol/L甘草酸处理MCF 7细胞 2 4h ,采用MTT比色法测定细胞增殖能力。 5 .0mmol/L、7.5mmol/L和 10 .0mmol/L甘草酸处理MCF 7细胞 2 4h ,用末端脱氧核苷转移酶介导dUTP末端标记法和AnnexinV流式细胞仪法检测凋亡细胞。 7.5mmol/L甘草酸处理MCF 7细胞 2 4h ,采用Fura 2荧光负载方法测定 [Ca2 ]i的变化。结果　甘草酸从 5 .0mmol/L浓度起对MCF 7细胞的增殖抑制率显著升高 (P <0 .0 1) ,呈剂量依赖性 ;半增殖抑制浓度 (IC50 )为 15 .8mmol/L。 7.5mmol/L和 10 .0mmol/L甘草酸使细胞凋亡率显著升高 (P <0 .0 5和P <0 .0 1)。甘草酸处理组的 [Ca2 ]i明显低于对照组 (P <0 .0 5 )。结论　甘草酸具有抑制MCF 7细胞增殖和诱导凋亡的作用 ;其诱导细胞凋亡可能与细胞内Ca2 水平下调有关     

18β-甘草次酸诱导人乳腺癌细胞凋亡及其细胞内Ca2+水平的变化

背景与目的：18-甘草次酸是甘草的重要成分，近年来的研究发现18-甘草次酸具有抑制人淋巴细胞性白血病、肝癌和肺癌的细胞增殖。本研究探讨18-甘草次酸对人乳腺癌MCF-7细胞诱导凋亡的作用。目前研究已证实细胞内游离Ca^2＋浓度（[Ca^2＋]i）的动态变化在诱发细胞凋亡过程的多个环节中起重要作用，因此本研究也探讨由18-甘草次酸诱导的MCF-7细胞凋亡发生与[Ca^2＋]i变化的关系。方法：用50～250μmol／L浓度梯度的18B-甘草次酸处理MCF-7细胞24h，用MTT比色法测定细胞增殖能力。100μmol／L和150pomol／L188-甘草次酸处理细胞24h，用末端脱氧核苷酸转移酶介导dUTP末端标记法、Annexin V流式细胞仪法和单细胞凝胶电泳法检测凋亡细胞。150μmol／L 18β-甘草次酸处理细胞24h，用Fura-2荧光负载方法测定[Ca^2＋]i的变化。分别用100μmol／LBAPTA-AM和0．5mmol／LEGTA与150μmol／L 18β-甘草次酸联合处理MCF-7细胞24h，用单细胞凝胶电泳法检测凋亡细胞。结果：从100μmol／L 18β-甘草次酸浓度起对MCF-7细胞的增殖抑制率显著升高（P〈0．01和P〈0．05），呈剂量依赖性，半增殖抑制浓度（IC50）为234．33μmol／L。100μmol／L和150μmol／L 18β-甘草次酸使细胞凋亡率显著升高（P〈0．01和P〈0．05）；18B-甘草次酸处理组的[Ca^2＋]i也明显高于对照组（P〈0．05）。18β-甘草次酸与BAPTA-AM联合处理组和18 β-甘草次酸与EGTA联合处理组的凋亡率均明显低于单纯的150μmol／L 18β-甘草次酸处理组（P〈0．05和P〈0．01）。结论：18-甘草次酸具有抑制MCF-7细胞增殖和诱导其凋亡的作用，而18-甘草次酸诱导MCF-7细胞凋亡的作用在抑制该细胞增殖方面起主要作用。18-甘草次酸诱导MCF-7细胞凋亡依赖于细胞内Ca^2＋水平上捌，而细胞外Ca^2＋内流是导致细胞内Ca^2＋水平上调的原因之一。     

18β-甘草次酸诱导人乳腺癌细胞凋亡及与细胞内Ca~(2+)释放的关系(英文)

目的探讨18β-甘草次酸对人乳腺癌细胞(MCF-7)增殖抑制和诱导凋亡的作用,并探讨凋亡诱发与细胞内 Ca~(2+)浓度([Ca~(2+)]i)变化的关系。方法 50～250 μmol/L 浓度梯度的18β-甘草次酸处理 MCF-7细胞24小时,用 MTF 比色法测定细胞增殖能力。100 μmol/L,150μmol/L 和200 μmol/L 18β-甘草次酸处理细胞24小时,用末端脱氧核苷转移酶介导 dUTP 末端标记法和 Annexin V 流式细胞仪法检测凋亡细胞。150 μmol/L 18β-甘草次酸处理细胞24小时,用 Fura-2荧光负载方法测定[Ca~(2+)]i 的变化。结果从100 μmol/L 18β-甘草次酸浓度起对 MCF-7细胞的增殖抑制率显著升高(P<0.05和 P<0.01),呈剂量依赖性,半增殖抑制浓度(IC_(50))为234.33 μmol/L.100 μmol/L、150 μmol/L 和200 μmol/L18β-甘草次酸使细胞凋亡率显著升高(P<0.01);18β-甘草次酸处理组的[Ca~(2+)]i 也明显高于对照组(P<0.05)。结论 18β-甘草次酸具有抑制 MCF-7细胞增殖和诱导凋亡的作用,其诱导凋亡可能依赖于细胞内 Ca~(2+)水平上调。     

18β-甘草次酸诱导人乳腺癌细胞凋亡及与细胞内Ca~(2 )释放的关系(英文)

目的 探讨18β-甘草次酸对人乳腺癌细胞(MCF-7)增殖抑制和诱导凋亡的作用,并探讨凋亡诱发与细胞内Ca2 浓度([Ca2 ]i)变化的关系。方法 50-250 μmol/L浓度梯度的18β-甘草次酸处理MCF-7细胞24小时,用MTT比色法测定细胞增殖能力。100 μmol/L,150μmol/L和200 μmol/L 18β-甘草次酸处理细胞24小时,用末端脱氧核苷转移酶介导dUTP末端标记法和Annexin V流式细胞仪法检测调亡细胞。150μmol/L 18β-甘草次酸处理细胞24小时,用Fura-2荧光负载方法测定[ca2 ]i的变化。结果 从100 μmol/L 18β-甘草次酸浓度起对MCF-7细胞的增殖抑制率显著升高(P<0.05和P<0.01),呈剂量依赖性,半增殖抑制浓度(IC50)为234.33 μmol/L。100 μmol/L、150 μmol/L和200 μmol/L18β-甘草次酸使细胞凋亡率显著升高(P<0.01);18β-甘草次酸处理组的[Ca2 ]i也明显高于对照组(P<0.05)。结论18β-甘草次酸具有抑制MCF-7细胞增殖和诱导凋亡的作用,其诱导凋亡可能依赖于细胞内Ca2 水平上调。     

Ca2+在鼻咽癌细胞凋亡性容积减小中的作用

 目的探讨钙信号途径在凋亡性细胞容积减小中的作用。方法用凋亡诱导剂顺铂诱导低分化鼻咽癌CNE-2Z细胞发生凋亡性细胞容积减小（AVD）,采用Q500MC软件控制定时拍摄固定视野活细胞图像,Scion Image图像分析软件检测细胞容积变化。结果细胞外灌流顺铂后细胞体积减小,10 min后细胞体积减小差异有统计学意义。去除灌流液中的Ca²⁺或用钙通道阻断剂Nifedipine阻断钙通道后,延缓顺铂诱导的细胞容积减小的发生,但不能防止细胞缩小过程的出现,50 min后,Nifedipine组和无Ca²⁺灌流液组与顺铂对照组AVD水平均无差异。结论细胞外Ca²⁺内流可能在触发细胞凋亡早期细胞容积减小中起重要作用。     

三氧化二砷诱导雌激素受体阴性乳腺癌细胞株MDA-MB-435s凋亡和bcl-2、bax表达关系的研究

目的 观察不同浓度三氧化二砷(As2O3)诱导人乳腺癌细胞系MDA-MB-435s凋亡的情况及对bcl-2和bax表达的影响,初步探讨了其诱导凋亡的途径.方法 不同浓度As2O3体外作用于MDA-MB-435s细胞24h后,采用TUNEL染色法和DNA梯状电泳法检测细胞凋亡情况,应用免疫组化法检测细胞中bcl-2和bax蛋白表达情况.结果 As2O3作用后,DNA琼脂糖凝胶电泳法呈现凋亡DNA梯状条带;As2O3作用组的凋亡指数明显高于阴性对照组(P＜0.05);bcl-2表达下调,bax表达上调.结论 As2O3能诱导MDA-MB-435s细胞凋亡,上调bcl-2蛋白表达、下调bax蛋白表达是其诱导凋亡的途径之一.     

基因促进TRAIL诱导乳腺癌MCF 7细胞的凋亡

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/extracellular signal regulated kinasekinase 3,MEKK 3)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转染入人乳腺癌细胞MCF7，以RTPCR和Western blotting法检测MCF7细胞MEKK 3 mRNA和蛋白的表达。应用MTT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 MEKK3 mRNA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。     

基因转染LL-37/hCAP-18对人乳腺癌细胞凋亡的影响

目的：探讨基因转染人源抗菌肽LL-37/hCAP-18对人乳腺癌细胞MCF-7增殖、凋亡的影响。方法：将含人LL-37/hCAP-18的真核表达质粒瞬时转染人MCF-7细胞,48h后,MTT比色检测MCF-7细胞的增殖,流式细胞术检测MCF-7细胞周期及凋亡,细胞免疫组化染色检测MCF-7细胞中Bax、Bcl-2的表达。结果：重组基因瞬时转染MCF-7细胞48h后,与各对照组相比,肿瘤细胞的生长增殖受到显著抑制（P〈0.05）;转染重组基因组肿瘤细胞凋亡率显著高于各对照组（P〈0.05）,但细胞周期无明显变化;与对照组相比,转染重组基因pEGFP-c1-LL-37的MCF-7细胞中Bcl-2表达显著下降（P〈0.05）,Bax由阴性表达转为表达明显升高。结论：LL-37/hCAP-18可以通过上调促凋亡因子Bax、下调抗凋亡因子Bcl-2诱导肿瘤细胞凋亡,从而抑制乳腺癌细胞MCF-7的增殖。     

多西紫杉醇诱导人乳腺癌MCF-7细胞凋亡及相关基因表达的影响

目的探讨多西紫杉醇(docetaxcel)诱导体外培养人乳腺癌MCF-7细胞凋亡的作用及凋亡的分子机制。方法通过MTT比色法分析多西紫杉醇对人乳腺癌MCF-7细胞增殖的影响;应用光镜、电镜观察细胞形态学的改变;Annexin-V/PI双染流式细胞术检测肿瘤细胞周期动力学和凋亡诱导率的影响;免疫组化方法检测凋亡相关基因bax、bc1-2的表达变化。结果多西紫杉醇可明显抑制人乳腺癌MCF-7细胞的增殖,并可导致其形态学的改变;随着加药时间的增长,G2/M期细胞增多,G2/M期细胞和S期细胞的比值也随剂量呈递增关系;加药(IC50)48h后早期凋亡细胞明显增多,同时可上调bax和下调bc1-2的蛋白表达量。结论多西紫杉醇可明显抑制MCF-7肿瘤细胞的生长,诱导细胞凋亡。其分子机制可能与激活bax和抑制bc1-2等凋亡调控基因有关。     

沙棘籽渣黄酮诱导人乳腺癌细胞凋亡相关基因的表达谱变化

背景与目的:研究显示,沙棘含有多种黄酮类化合物,具有抗氧化、防辐射等作用。cDNA基因表达谱芯片技术具有高通量、高效、快捷的特点,本实验拟利用cDNA基因表达谱芯片研究沙棘籽渣黄酮(flavonoidsfromseedresiduesofHippophaerhamnoidesL.,FHR)诱导人乳腺癌细胞Bcap鄄37凋亡相关基因的表达谱变化。方法:抽提FHR作用前后的Bcap鄄37细胞总RNA,经逆转录合成荧光分子(Cy3/Cy5)标记的cDNA探针,与含有13824个cDNA基因的人14K基因表达谱芯片杂交,利用Genespring软件分析实验组和对照组之间差异表达的基因,对所获得的基因进行生物信息学分析。结果:实验组与对照组中表达上调的基因有305条,表达下调的基因有361条。差异表达的与细胞凋亡相关的基因有32条,占芯片基因总数的0.23%,其中表达上调的有25条(平均Ratio值为3.071),下调的有7条(平均Ratio值为0.418)。生物信息学分析表明,FHR在对Bcap鄄37细胞作用的过程中,该32条差异表达基因与Bcap鄄37细胞凋亡有关,其中包括CTNNB1、TSSC3、IGFBP4、IGFBP6、GADD34、TNFRSF10B、Caspase鄄9和PCNA等。结论:FHR诱导Bcap鄄37细胞凋亡的机制是由多基因协同,并通过胞内和胞外信号转导途径共同调节完成。     

Tamoxifen-Induced Increases in Cytoplasmic Free Ca2+ Levels in Human Breast Cancer Cells

Tamoxifen has been shown to increase cytoplasmic free Ca²⁺ levels [Ca²⁺]_i in renal tubular cells and bladder cancer cells, and to alter Ca²⁺ signaling in MCF-7 breast cancer cells. The present study examined the effect of tamoxifen on [Ca²⁺]_i in ZR-75-1 human breast cancer cells using fura-2 as an indicator. Tamoxifen increased [Ca²⁺]_i at a concentration above 2M with an EC₅₀ of 5M. Removing extracellular Ca²⁺ reduced the response by 48±2%. In Ca²⁺-free medium, after tamoxifen-induced [Ca²⁺]_i increased had returned to baseline, adding 3mM Ca²⁺ increased [Ca²⁺]_i in a concentration-dependent manner. Further, pretreatment with 10M tamoxifen abolished the [Ca²⁺]_i increase induced by 1M thapsigargin (an endoplasmic reticulum Ca²⁺ pump inhibitor); and conversely, pretreatment with thapsigargin prevented tamoxifen from releasing more Ca²⁺. Tamoxifen (10M)-induced Ca²⁺ release was not changed by inhibiting phospholipase C activity with 2M U73122. Trypan blue exclusion assay revealed that tamoxifen (1–10M) did not alter viability after 1min of incubation, but killed 10% of cells after 3–10min of incubation. Together, this study shows that tamoxifen (>2M) induced a significant, immediate increase in [Ca²⁺]_i in ZR-75-1 breast cancer cells. Tamoxifen acted by releasing Ca²⁺ from the endoplasmic reticulum Ca²⁺ stores in a manner independent of phospholipase C activity, and by inducing Ca²⁺ entry from extracellular medium. Tamoxifen may be of mild cytotoxicity after acute exposure.     

Apoptosis of Human Ovarian Cancer Cells Induced by Paris Chinensis Dioscin via a Ca2+-Mediated Mitochondrion Pathway

Background: Study of the mechanisms of apoptosis in tumor cells is an important field of tumor therapy andcancer molecular biology. Apoptosis triggered by activation of the mitochondrial-dependent caspase pathwayrepresents the main programmed cell death mechanism. The mitochondrial-dependent apoptosis pathway isactivated by various intracellular stresses that induce permeabilization of the mitochondrial membrane, leadingto cytochrome C release. This study was to investigate the anti-tumor effects of Dioscin from traditional Chineseanti-snake venom medicine Paris chinensis (PCD) and correlated mechanisms regarding apoptosis in humanovarian cancer SKOV3 cells. Methods: Cell viability was analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Cell apoptosis was evaluated by flow cytometry and Laser ScanningConfocal Microscope (LSCM) using Annexin-V/PI staining. Intracellular calcium ions were detected usingfluorescence microscopy. The expression of apoptosis-related proteins cytochrome C and caspase-3 was measuredby immunohistochemical staining. Results: PCD had an anti-proliferation effect on human ovarian cancerSKOV3 cells in a dose- and time-dependent manner. After treatment with PCD, the apoptotic rate significantlyincreased, and accompanied with the increased levels of caspase-3 and cytochrome C protein in SKOV3 cells.Morphological changes typical of apoptosis were also observed with LSCM by Annexin V/PI staining. Moreover,intracellular calcium accumulation occurred in PCD-treated cells. Conclusions: The molecular determinants ofinhibition of cell proliferation as well as apoptosis of PCD may be associated with the activation of Ca2+-relatedm itochondrion pathway in SKOV3 cells.     

吉非替尼对人乳腺癌细胞增殖和凋亡的影响

[目的]探讨Iressa单药及联合化疗对乳腺癌细胞MCF-7及MCF-7/ADR的抑制作用。[方法]用MTT法测定两种细胞系在应用Iressa及联合化疗药物后的细胞抑制率。利用流式细胞技术检测细胞周期分布。[结果]光镜观察到,MCF-7及MCF-7/ADR在分别加入Iressa及TE后细胞形态不规则,细胞内结构消失。MTT数据显示MCF-7和MCF-7/ADR细胞的抑制率与Iressa浓度成正相关,随Iressa浓度升高,抑制作用越明显;联合TE与单用Iressa相比不能提高对乳腺癌细胞MCF-7的细胞抑制率(P>0.05),与单用TE比能明显提高对乳腺癌细胞MCF-7的细胞抑制率(P<0.01);但联合用药与单用比均能明显提高对MCF-7/ADR的细胞抑制率(P<0.01)。流式细胞术分析显示经Iressa处理24h MCF-7和MCF-7/ADR细胞G0～G1期细胞比例明显升高,且随浓度升高而升高明显。[结论]靶向药物Iressa对MCF-7及MCF-7/ADR细胞均有抑制作用,联合应用对MCF-7和MCF-7/ADR的抑制无协同作用,但考虑Iressa对MCF-7/ADR有逆转耐药的作用。联合TE中的给药顺序对细胞的抑制率无明显影响。     

熊果酸对人乳腺癌细胞增殖、凋亡和细胞内游离Ca2+的影响

[目的]探讨熊果酸对人乳腺癌细胞(MCF-7)增殖抑制和诱导凋亡的作用,以及凋亡发生与细胞内Ca2 浓度([Ca2 ]i)变化的关系.[方法]5～40μmol/L浓度梯度的熊果酸处理MCF-7细胞24小时,用MTT比色法测定细胞增殖能力.10μmol/L,20μmol/L和30μmol/L熊果酸处理细胞24小时,用末端脱氧核苷转移酶介导dUTP末端标记法和Annexin V流式细胞仪法检测凋亡细胞.20μmol/L熊果酸处理细胞24小时,用Fura-2荧光负载方法测定[Ca2 ]i的变化.[结果]从20μmol/L熊果酸浓度起对MCF-7细胞的增殖抑制率显著升高(P＜0.01),呈剂量依赖性,半增殖抑制浓度(IC50)为36.18μmol/L.20μmol/L和30μmol/L熊果酸使细胞凋亡率显著升高(P＜0.01).熊果酸处理组的[Ca2 ]i明显高于对照组(P＜0.05).[结论]熊果酸具有抑制MCF-7细胞增殖和诱导凋亡的作用;其诱导凋亡可能依赖于细胞内Ca2 水平上调.     

Silibinin Enhances Ultraviolet B-Induced Apoptosis in MCF-7 Human Breast Cancer Cells

Purpose

Chemotherapies for breast cancer generally have strong cellular cytotoxicity and severe side effects. Thus, significant emphasis has been placed on combinations of naturally occurring chemopreventive agents. Silibinin is a major bioactive flavonolignan extracted from milk thistle with chemopreventive activity in various organs including the skin, prostate, and breast. However, the mechanism underlying the inhibitory action of silibinin in breast cancer has not been completely elucidated. Therefore, we investigated the effect of silibinin in MCF-7 human breast cancer cells and determined whether silibinin enhances ultraviolet (UV) B-induced apoptosis.

Methods

The effects of silibinin on MCF-7 cell viability were determined using the MTT assay. The effect of silibinin on PARP cleavage, as the hallmark of apoptotic cell death, and p53 protein expression in MCF-7 cells was analyzed using Western blot. The effect of silibinin on UVB-induced apoptosis in MCF-7 cells was analyzed by flow cytometry.

Results

A dose- and time-dependent reduction in viability was observed in MCF-7 cells treated with silibinin. Silibinin strongly induced apoptotic cell death in MCF-7 cells, and induction of apoptosis was associated with increased p53 expression. Moreover, silibinin enhanced UVB-induced apoptosis in MCF-7 cells.

Conclusion

Silibinin induced a loss of cell viability and apoptotic cell death in MCF-7 cells. Furthermore, the combination of silibinin and UVB resulted in an additive effect on apoptosis in MCF-7 cells. These results suggest that silibinin might be an important supplemental agent for treating patients with breast cancer.     

Fentanyl Inhibits Tumorigenesis from Human Breast Stem Cells by Inducing Apoptosis

Fentanyl is an opioid analgesic that it is widely used in cancer patients. Since there have been reports of effects of analgesic medications on the recurrence and development of resistance to treatment, influences of of fentanyl on MCF-7 and HEK293 cells were evaluated. Cell viability and apoptosis were assessed by MTT assay and flow cytometry, respectively. Gene expression analysis was performed by quantitative real-time PCR assay for the Oct4, Sox2 and Nanog genes as stem cell markers and Bax, Bcl2, and p53 genes as apoptosis markers. MTT assay results showed that fentanyl significantly inhibited the growth of MCF-7 cells in a dose-and time-dependent manner while significantly increasing apoptosis. In contrast, decrease was noted in HEK-293 cells. In MCF-7 derived cancer stem cells, fentanyl treatment decreased the expression of Bax, Bcl2, Oct4, Sox2, Nanog genes when compared to untreated cells. In HEK-293 stem cells, decrease was noted for Sox2, Nanog and Bax, but increase for Oct4. Our study supports an antitumor role of fentanyl by inducing apoptosis and reducing numbers of cancer stem cells in the MCF-7 human breast adenocarcinoma line.     

骨形成蛋白-2(BMP-2)与雌激素敏感性的人类乳腺癌的关系

目的：研究BMP-2(Bone Morphogenetic Protein-2，骨形成蛋白-2)在乳腺癌细胞系中的表达，探讨其对乳腺癌细胞的作用机制，评估其在乳腺癌基因治疗上的作用。方法：采用分子生物学方法（Molecular Biology）检测6种人类乳腺癌细胞系中BMP-2的表达，并分析对乳腺癌细胞生长的影响及其与治疗的关系。结果：BMP-2在所有人类乳腺癌细胞系中表达，其抑制乳腺癌细胞的生长，主要抑制于细胞周期的G1期，其抑制作用与p21蛋白的诱导有关。结论：BMP-2是对雌激素敏感的人类乳腺癌细胞的强有力抑制剂，这有可能成为在分子水平上治疗乳腺癌的新方法。     

乳腺肿瘤基质金属蛋白酶2表达与肥大细胞的关系

[目的]探讨乳腺肿瘤中基质金属蛋白酶2(matrix meta]]oproteinase2,MMP-2)表达及其与肥大细胞浸润的关系.[方法]采用甲苯胺蓝染色显示55例良恶性乳腺肿瘤组织的肥大细胞,免疫组化EnVision法检测肿瘤细胞MMP-2以及类胰蛋白酶(tryptase)的表达.[结果]乳腺癌间质成纤维细胞MMP-2的反应程度明显高于癌细胞.浸润性小叶癌和浸润性导管癌MMP-2的阳性率达86.7%,明显高于恶性程度较低的黏液腺癌和髓样癌(20.0%)以及乳腺的良性肿瘤(13.3%).局部有转移的乳腺癌组织中MMP-2的表达程度明显高于无转移组.乳腺肿瘤间质中肥大细胞的数量以及tryptase的表达量与MMP-2相似,但是与MMP-2的表达量并无明显的相关性.[结论]MMP-2的表达是乳腺癌一个预后指标,而肥大细胞的增多可能是肿瘤纤维间质形成的相伴现象.     

Isocryptotanshinone Induced Apoptosis and Activated MAPK Signaling in Human Breast Cancer MCF-7 Cells

Purpose

Isocryptotanshinone (ICTS) is a natural bioactive product that is isolated from the roots of the widely used medical herb Salvia miltiorrhiza. However, few reports exist on the mechanisms underlying the therapeutic effects of ICTS. Here, we report that ICTS has anticancer activity and describe the mechanism underlying this effect.

Methods

The antiproliferative effect of ICTS was determined using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and clonogenic assays. The effect of ICTS on the cell cycle was measured using flow cytometry. Apoptosis was determined by Hoechst 33342 staining, DNA fragmentation assays, and Western blotting for apoptotic proteins. Finally, the effect of ICTS on mitogen-activated protein kinases (MAPKs) was determined by Western blotting.

Results

ICTS significantly inhibited proliferation of MCF-7 and MDA-MB-231 human breast cancer cells, HepG2 human liver cancer cells, and A549 human lung cancer cells in vitro. Among the tested cell lines, MCF-7 cells showed the highest sensitivity to ICTS. ICTS significantly inhibited colony formation by MCF-7 cells. Furthermore, exposure of MCF-7 cells to ICTS induced cell cycle arrest at the G1 phase and decreased mitochondrial membrane potential. Hoechst 33342 staining and Western blot analysis for apoptotic proteins suggested that ICTS induced apoptosis in MCF-7 cells. In addition, ICTS activated MAPK signaling in MCF-7 cells by inducing time- and concentration-dependent phosphorylation of JNK, ERK, and p38 MAPK.

Conclusion

Our results suggest that ICTS inhibited MCF-7 cell proliferation by inducing apoptosis and activating MAPK signaling pathways.