Bone morphogenetic protein signaling in breast cancer progression

Breast cancer is the most prevalent type of cancer amongst women worldwide. The mortality rate for patients with early-stage breast cancer has been decreasing, however, the 5-year survival rate for patients with metastatic disease remains poor, currently at 27%. Here, we have reviewed the current understanding of the role of bone morphogenetic protein (BMP) signaling in breast cancer progression, and have highlighted the discordant results that are reported in different studies. We propose that some of these contradictory outcomes may result from signaling through either the canonical or non-canonical pathways in different cell lines and tumors, or from different tumor-stromal interactions that occur in vivo.     

体外骨形态发生蛋白4抑制乳腺癌细胞的增殖和迁移

目的:研究骨形态发生蛋白4(BMP4)对乳腺癌细胞MCF-7增殖、迁移和侵袭能力的影响.方法:将重组慢病毒Lv-BMP4及干扰慢病毒Lv-shBMP4感染MCF-7细胞,构建MCF-7/Lv-BMP4过表达组及MCF-7/Lv-shBMP4干扰实验组,同时设空白组(Blank)、慢病毒空载对照组(Lv-NC)、 过表达...     

骨形态发生蛋白9调控干细胞分化与骨再生

背景:骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)具有很强的促进干细胞成骨分化的潜能.大量研究表明,BMP9可在动物体内促进骨缺损的修复.目的:探究BM P9调控干细胞分化的机制,讨论影响BMP9促进干细胞成骨分化的潜在因素以及BMP9在骨组织工程中的应用前景.方法:在PubM...     

Bone morphogenetic protein 7 is widely overexpressed in primary breast cancer

Bone morphogenetic proteins (BMP) make up a family of extracellular signaling molecules that play a critical role in vertebrate development and both inhibit and stimulate growth in cancer cells. BMP7 was recently identified in our genomewide copy number and expression survey as being activated through amplification in breast cancer cell lines. In the present study, we further explored BMP7 gene copy number and expression changes in 22 breast cancer cell lines and 146 primary breast tumors. FISH analysis revealed that BMP7 copy number varied greatly from one cell line to another, with three cell lines showing extremely high-level amplification. Among primary tumors, BMP7 copy number was increased in 16% of the cases. BMP7 mRNA expression was determined in the cell lines and in a subset of 44 tumor samples by RT-PCR or quantitative real-time RT-PCR, respectively. Despite elevated mRNA levels in cancer cells, there was no significant association between copy number increase and mRNA expression, even though the highest expression was seen in cell lines and tumors with increased BMP7 copy number. Most interestingly, immunohistochemical analysis revealed BMP7 protein staining in all 11 breast cancer cell lines examined and strongly elevated BMP7 protein expression in 71.4% of the tumor samples as compared to normal mammary epithelium. Our results illustrate the frequent involvement of BMP7 alterations in breast cancer and especially highlight overexpression of the BMP7 protein in a very large fraction of primary breast tumors, thus suggesting a possible functional role for BMP7 in breast cancer development.     

骨形态发生蛋白9体外调节乳腺癌细胞与骨髓间充质干细胞的相互作用

 目的 观察骨形态发生蛋白9（BMP9）在共培养条件下对HS-5成骨分化及MDA-MB-231转移相关因子表达的影响。方法 采用Transwell小室间接共培养MDA-MB-231细胞与HS-5细胞,加入BMP9条件培养基,使用化学发光法、细胞化学染色法及茜素红S染色法分别检测HS-5细胞碱性磷酸酶（ALP）活性、ALP表达及细胞钙盐沉积的变化;应用RT-PCR及Western blot观察HS-5细胞骨钙素（OCN）、MDA-MB-231细胞甲状旁腺激素相关蛋白（PTH-rP）及白细胞介素6(IL-6)mRNA和蛋白表达的变化。结果 HS-5的ALP活性升高,并呈时间依赖性,第9天达最高,随后降低;ALP细胞化学染色结果进一步证实这一点;HS-5细胞钙盐沉积增多、OCN mRNA和蛋白的表达升高（P<0.05）;MDA-MB-231转移相关的PTH-rP及IL-6 mRNA和蛋白的表达下降（P<0.05）。结论 BMP9能够调节乳腺癌细胞与骨髓间充质干细胞的相互作用。     

Bone morphogenetic protein signaling regulates skin inflammation via modulating dendritic cell function

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Bone morphogenetic protein signaling regulates postnatal hair follicle differentiation and cycling

Hair follicle morphogenesis and cycling were examined in transgenic mice that overexpress the bone morphogenetic protein (BMP) inhibitor Noggin under the control of the neuron-specific enolase promoter. The Noggin transgene was misexpressed in the proximal portion of the hair follicle, primarily the matrix cells, apart from the usual expression in neurons. Transgene expression appeared only after induction of both the primary (tylotrich) and secondary (nontylotrich) pelage hair follicles had already occurred, thus allowing examination of the role of BMP signaling in follicles that had been induced normally in the presence of BMPs. The overexpression of Noggin in these animals resulted in a dramatic loss of hair postnatally. There was an apparently normal, but shortened period of postnatal hair follicle morphogenesis, followed by premature initiation of hair follicle cycling via entry into the first catagen transformation. This resulted in a complete loss of hair shafts from the nontylotrich hair follicles in these mice while the tylotrich hair follicles were normal. The onset of anagen of the first postnatal hair follicle cycle was also accelerated in the transgenic mice. Our results show that BMP signaling is specifically required for proper proliferation and differentiation during late morphogenesis of nontylotrich hair follicles and that inhibition of this signaling pathway may be one of the triggers for the onset of catagen when the follicles are in anagen and the onset of anagen when the follicles are in telogen. Ectopic sebocyte differentiation was another hallmark of the phenotype of these transgenic mice suggesting that BMP signaling may be an important determinant of lineage selection by common progenitor cells in the skin. BMPs likely promote a hair follicle-type differentiation pathway of keratinocytes while suppressing the sebaceous differentiation pathway of skin epithelium.     

人结肠癌细胞系SP细胞microRNA表达谱的初步分析

 目的：探讨结肠癌侧群（SP）细胞在结肠癌多药耐药性中的作用及其microRNA生物标志。方法：结肠癌SP细胞的分选采用流式细胞术,细胞活力的测定采用MTT法,microRNA表达谱的检测采用microRNA芯片,microRNA表达的验证采用实时荧光定量PCR。结果：（1）HCT-15、HT-29及LoVo结肠癌细胞系中SP细胞的比例分别为16.75%、13.02%及9.52%。（2） 化疗药（5-氟尿嘧啶、草酸铂及阿霉素）对3种结肠癌细胞系SP细胞的 IC50均明显高于对非SP细胞的IC50（P<0.05）。（3）microRNA芯片检测表明miR-5000-3p、miR-5009-3p及miR-552在3种结肠癌细胞系的SP细胞中表达均上调,实时荧光定量PCR亦证实此结果。结论：miR-5000-3p、miR-5009-3p及miR-552在结肠癌细胞系的SP细胞中表达上调,有可能成为结肠癌SP细胞的潜在microRNA生物标志。     

Metronidazole affects breast cancer cell lines

PurposeThe aim of our study was to evaluate the impact of metronidazole (MTZ) on cytotoxicity and DNA synthesis in MCF-7 (estrogen receptor positive) and MDA-MB-231 (estrogen receptor negative) breast cancer cell lines.Material/MethodsToxicity of MTZ was determined by MTT test. MCF-7 and MDA-MB-231 cells were incubated with metronidazole used in different concentrations for 24, 48 and 72 hours. The effect of MTZ on DNA synthesis was measured as [3H]-thymidine incorporation.ResultsWe showed that MTZ in concentration 250 μg/ml significantly increases the growth of MCF-7 cell lines after 24 hours of incubation, but it reduces cell viability in concentrations 1 and 10 μg/ml 72 hours after the drug application. Significant increase of MDA-MB-231 cell viability was obtained in MTZ concentration of 250 μg/ml after 24 and 72 hours. The increase of [3H]-thymidine incorporation in MCF-7 cell line treated with MTZ in concentration 250 μg/ml was statistically significant after 24 hours. Great suppression of cell proliferation was obtained in MDA-MB-231 breast cell line after application of the following concentrations of MTZ: 0.1 μg/ml (after 24 hours) and 0.1, 10, 50, 250 μg/ml (after 72h).ConclusionsWe found that metronidazole exerts different dose- and time- dependent effects on human breast cancer cell lines characterized by presence or absence of estrogen receptors. We suggest that these discrepancies may be influenced by the estrogen signaling.     

Effect of histamine on the expression of metalloproteinases and cell adhesion in breast cancer cell lines

Inflammation Research -     

Y14和Upf1在人乳腺癌细胞表达增强

目的 探讨Y14和Upf1在人乳腺癌细胞株和人乳腺癌组织表达情况及其意义。方法 应用免疫细胞化学、激光共聚焦方法测定Y14和Upf1在乳腺癌细胞株MCF-7、ZR-75-30、T47D、MDA-MB-435s、MDA-MB-453、MDA-MB-231与乳腺上皮细胞株HBL-100中的表达情况。结果 ⑴人乳腺癌细胞株中Y14和Upf1的表达均明显高于乳腺上皮细胞株（P＜0.05）。⑵在人乳腺癌细胞中,MDA-MB-231细胞株的Y14和Upf1表达明显高于MCF-7细胞（P＜0.05）。⑶Y14和Upf1在人乳腺癌组织表达增强（P＜0.05）。结论 Y14和Upf1在人乳腺癌细胞和人乳腺癌组织中的表达增强。     

Botulinum neurotoxin type A inhibits synaptic vesicle 2 expression in breast cancer cell lines

Aim: It is known that botulinum neurotoxin type A (BoNTA) improves some kinds of cancer (e.g. prostate) and that synaptic vesicle glycoprotein 2 (SV2) is the molecular target of this neurotoxin. Besides having potential therapeutic value, this glycoprotein has recently been proposed as a molecular marker for several types of cancer. Although the mechanisms of cancer development and the improvement found with botulinum treatment are not well understood, the formation of the botulinum-SV2 complex may influence the presence and distribution of SV2 and the function of vesicles. To date, there are no reports on the possible effect of botulinum on breast cancer of unknown causes, which have a great impact on women’s health. Thus we determined the presence of SV2 in three breast cancer cell lines and the alterations found with botulinum application. Materials and methods: With and without adding 10 units of botulinum, SV2 protein expression was determined by optical densitometry in T47D, MDA-MB-231 and MDA-MB-453 cell lines and the distribution of SV2 was observed with immunochemistry (hematoxylin staining). Results: The SV2 protein was abundant in the cancer cells herein tested, and maximally so in T47D. In all three cancer cell lines botulinum diminished SV2 expression, which was found mostly in the cell periphery. Conclusion: SV2 could be a molecular marker in breast cancer. Its expression and distribution is regulated by botulinum, suggesting an interesting control mechanism for SV2 expression and a possible alternative therapy. Further studies are needed in this sense.     

不同长度兔桡骨缺损区骨形态发生蛋白2的表达

背景：有研究表明在骨缺损的状态下,骨形态发生蛋白2的分布存在一定的规律。 目的：观察不同长度兔桡骨骨缺损部位骨形态发生蛋白2的表达。 方法：将48只新西兰大白兔随机分成2组,分别在麻醉下用线锯造成兔左前臂桡骨中段0.5和3.0 cm的骨缺损。 结果与结论：Western blot检测显示,0.5 cm骨缺损组骨缺损部位骨形态发生蛋白2蛋白表达量在损伤后1,3,4周呈逐渐增多的趋势,且每组均较损伤即刻明显增加(P < 0.05);3.0 cm骨缺损组骨缺损部位的骨形态发生蛋白2的相对表达量在损伤后3周达到高峰(P < 0.05),其峰值水平明显高于0.5 cm骨缺损组(P < 0.05),基本维持在较高水平。对骨痂的生成情况进行量化评估结果显示,损伤后3,4周,3.0 cm骨缺损组的骨痂生成量较0.5 cm骨缺损组明显减少(P < 0.05)。证实,兔桡骨3.0 cm的骨缺损部位骨形态发生蛋白2表达明显上调,但还不能使该长度的骨缺损自行愈合。     

Bone morphogenetic protein 4 inhibits liposaccharide‐induced inflammation in the airway

Bone morphogenetic protein 4 (BMP4) is a multifunctional growth factor that belongs to the TGF‐β superfamily. The role of BMP4 in lung diseases is not fully understood. Here, we demonstrate that BMP4 was upregulated in lungs undergoing lipopolysaccharide (LPS)‐induced inflammation, and in airway epithelial cells treated with LPS or TNF‐α. BMP4 mutant (BMP4^+/?) mice presented with more severe lung inflammation in response to LPS or Pseudomonas aeruginosa, and lower bacterial load compared with that in BMP4^+/+ mice. Knockdown of BMP4 by siRNA increased LPS and TNF‐α‐induced IL‐8 expression in 16HBE human airway epithelial cells and in primary human bronchial epithelial cells. Similarly, peritoneal macrophages from BMP4^+/? mice produced greater levels of TNF‐α and keratinocyte chemoattractant (KC) upon LPS treatment compared with cells from BMP4^+/+ mice. Administration of exogenous BMP4 attenuated the upregulation of TNF‐α, IL‐8, or KC induced by LPS and/or TNF‐α in airway epithelial cells, and peritoneal macrophages. Finally, partial deficiency of BMP4 in BMP4^+/? mice protected the animals from restrictive lung function reduction upon chronic LPS exposure. These results indicate that BMP4 plays an important anti‐inflammatory role, controlling the strength and facilitating the resolution of acute lung inflammation; yet, BMP4 also contributes to lung function impairment during chronic lung inflammation.     

Bone morphogenetic protein 2 induces the expression of cementum attachment protein in human periodontal ligament clones

Cementum is continuously formed during the lifetime of a tooth. The paravascular zones in the adult periodontal ligament (PL) comprise the progenitors for the fibroblastic (Fb) lineage and mineralized tissue-forming (MTF) cell lineages--the osteoblastic (Ob) and cementoblastic (Cb) lineages. Recent studies indicate that cementum attachment protein (CAP) is related to the differentiation of the Cb lineage and is instrumental in differentiating between the three periodontal cell lineages. The purpose of this study was to assess the effect of bone morphogenetic protein 2 (BMP2) on the expression of cementum attachment protein (CAP) and on the differentiation of cloned PL progenitors. The effect of BMP2 on CAP expression and on the differentiation of cloned Fb and MTF progenitors was tested by assessing the expression of alkaline phosphatase (ALP), CAP, and bone sialoprotein (BSP) by immunochemistry and by determining the CAP-binding capacity of these clones. Untreated Fb clones were negative for all tested markers and had low CAP-binding capacity. Untreated MTF clones had a high CAP-binding capacity and were positive for the three markers. BMP2 enhanced the CAP-binding potential of both Fb and MTF clones. BMP2 induced the expression of CAP, ALP, and BSP in the Fb clones and enhanced the expression of CAP and BSP in the MTF clones. These results indicate for the first time that BMP2 can recruit progenitors to the Cb lineage and regulate the differentiation of the Cb lineage by inducing and enhancing the expression of CAP, a cell lineage-specific regulator. Furthermore, the results suggest that the MTF and Fb lineages may originate from a common early progenitor cell.     

乳腺癌组织及细胞系中CCDC34的表达及其生物学功能

目的 探索乳腺癌组织及细胞系中CCDC34的表达情况及其对乳腺癌细胞增殖能力的影响。方法免疫组织化学染色方法检测34例乳腺癌组织及15例癌旁正常乳腺组织中CCDC34的表达和定位情况;利用Kaplan-Meier Plotter和The Human Protein Atlas数据库分析CCDC34与乳腺癌患者预后的相关性;Western blot检测人正常乳腺细胞MCF-10A及乳腺癌细胞SKBR-3、MCF-7、MDA-MB-453和MDA-MB-231中CCDC34的表达情况;在MCF-7和MDA-MB-231细胞中瞬时转染CCDC34,使用MTT的方法检测CCDC34对乳腺癌细胞增殖能力的影响。结果 CCDC34在乳腺癌组织的表达显著低于癌旁正常乳腺组织（P<0.01）;CCDC34在乳腺癌中的表达水平与患者的不良预后显著负相关（P<0.01）;CCDC34在乳腺癌细胞中的表达普遍低于人正常乳腺细胞MCF-10A(P<0.01);MTT检测结果显示,转染CCDC34显著抑制了乳腺癌细胞MCF-7(Luminal A)和MDA-MB-231(basal)细胞的增殖能力（P<0.05）。结论 乳腺癌组织及细胞系中CCDC34的表达普遍降低,CCDC34能够抑制乳腺癌细胞的增殖能力,并与乳腺癌患者的良好预后相关。     

脐带来源间充质干细胞在成脂培养条件下骨形态发生蛋白2的表达

背景：脐带间充质干细胞在成脂培养条件下成脂相关信号因子过氧化物酶体增殖体激活受体γ的表达水平增高,同时成骨相关信号因子骨形态发生蛋白的表达水平也增高。 目的：检测成脂诱导条件下脐带间充质干细胞的过氧化物酶体增殖体激活受体γ及骨形态发生蛋白2的表达水平变化,并对结果进行初步探讨。 方法：取脐带来源的间充质干细胞,以成脂诱导体系对细胞进行诱导分化培养,倒置显微镜下观察细胞体外培养生长情况;油红O染色法观察成脂现象,茜素红及von kossa染色观察是否有沉淀形成;荧光定量PCR方法检测成脂相关因子过氧化物酶体增殖体激活受体γ及骨形态发生蛋白2的表达变化情况。 结果与结论：获得脐带来源间充质干细胞,并成功进行了成脂诱导分化。荧光定量PCR检测发现在成脂诱导条件下脐带间充质干细胞的过氧化物酶体增殖体激活受体γ和骨形态发生蛋白2的表达水平全都呈升高趋势,但是前者对分化方向的调控起主要作用。     

乳腺癌发生模型MCF10 抑癌基因NOEY2启动子区甲基化及mRNA表达

DNA甲基化是常见的表观遗传学变化。越来越多的证据表明,抑癌基因启动子区CpG岛甲基化可使其转录沉默,从而解除其抑癌作用,导致肿瘤的发生和发展。NOEY2是近年来发现的抑癌基因,表达于正常乳腺导管上皮细胞和卵巢生发上皮细胞,但在乳腺癌和卵巢癌细胞中表达显著减少或不表达。在本实验中检测乳腺癌发生模型MCF10中不同阶段的细胞系NOEY2基因启动子区CpG岛Ⅰ甲基化状态,并与mRNA表达水平进行比较,以研究乳腺癌的发生机制和早期诊断。