目的探讨血管内皮生长因子(VEGF)对三阴性乳腺癌肿瘤干细胞的调控作用并探索其作用机制。方法通过Oncomine数据库、UALCAN数据库及Human Protein Atlas数据库分别验证VEGF在乳腺癌中的表达情况,分析VEGF表达与不同分子亚型的关系,利用...目的探讨血管内皮生长因子(VEGF)对三阴性乳腺癌肿瘤干细胞的调控作用并探索其作用机制。方法通过Oncomine数据库、UALCAN数据库及Human Protein Atlas数据库分别验证VEGF在乳腺癌中的表达情况,分析VEGF表达与不同分子亚型的关系,利用在线数据库分析VEGF的表达情况与乳腺癌预后的关系。选取三阴性乳腺癌MDA-MB-231细胞,加入外源性hVEGF165通过体外微球形成实验观察体外成球能力,并利用Western blot、RT-qPCR等方法检测肿瘤干细胞相关标志物CD44、c Myc、Nanog、ALDH1的表达以及相关通路的激活情况。结果利用Oncomine、UALCAN、HPA数据库分析显示,VEGF在乳腺癌中表达较癌旁组织增高(P<0.0001),其表达与分子亚型相关,三阴性乳腺癌中VEGF表达显著升高。Kaplan-Meier Plotter数据库分析结果提示高表达VEGF的乳腺癌患者预后更差,总生存期OS缩短,差异具有统计学意义(P<0.0001)。与VEGF低表达患者相比,VEGF高表达的乳腺癌患者无进展生存期、无远处转移生存期以及复发后生存期均显著缩短,差异具有统计学意义(P<0.0001)。体外实验发现加入外源性的hVEGF165后三阴性乳腺癌MDA-MB-231细胞的成球能力显著增强,差异具有统计学意义(P=0.0029)。Western blotting、RT-qPCR结果表明乳腺癌MDA-MB-231微球体中VEGF/NRP-1及肿瘤干细胞相关标志物CD44、Nanog、c-Myc的表达量显著升高。加入外源性的hVEGF165促进了肿瘤干细胞相关标志物CD44、c-Myc、Nanog及ALDH1的表达,而CD24的表达量则显著下降,上述作用具有时间依赖性。Western blotting实验提示加入外源性的hVEGF165能够激活三阴性乳腺癌细胞ERK/MAPK通路。结论VEGF可能通过激活ERK/MAPK通路促进了三阴性乳腺癌肿瘤干细胞的形成。展开更多
BACKGROUND Breast cancer is a common malignant tumor that seriously threatens women’s health.Breast cancer stem cell(CSC)-like cell population may be the main factor for breast cancer metastasis.Therefore,targeted th...BACKGROUND Breast cancer is a common malignant tumor that seriously threatens women’s health.Breast cancer stem cell(CSC)-like cell population may be the main factor for breast cancer metastasis.Therefore,targeted therapy for CSCs has great potential significance.Hypoxia-inducible factor is a transcription factor widely expressed in tumors.Studies have shown that down-regulation of the hypoxia signaling pathway inhibits tumor stem cell self-renewal and increases the sensitivity of stem cells to radiotherapy and chemotherapy mediated by hypoxiainducible factor-2α(HIF-2α).However,the specific mechanism remains unclear and further research is necessary.AIM To investigate the effect of HIF-2αdown-regulation on stem cell markers,microsphere formation,and apoptosis in breast cancer cell line MDA-MB-231 under hypoxia and its possible mechanism.METHODS Immunohistochemistry was used to detect the expression of HIF-2αand CD44 in triple-negative breast cancer(TNBC)and non-TNBC tissues.Double-labeling immunofluorescence was applied to detect the co-expression of HIF-2αand CD44 in MDA-MB-231 cells and MCF-7 cells.HIF-2αwas silenced by RNA interference,and the expression of CD44 and transfection efficiency were detected by real-time fluorescent quantitative PCR.Further,flow cytometry,TdT-mediated X-dUTP nick end labeling,and mammosphere formation assays were used to evaluate the effect of HIF-2αon CSCs and apoptosis.The possible mechanisms were analyzed by Western blot.RESULTS The results of immunohistochemistry showed that HIF-2αwas highly expressed in both TNBC and non-TNBC,while the expression of CD44 in different molecular types of breast cancer cells was different.In in vitro experiments,it was found that HIF-2αand CD44 were expressed almost in the same cell.Compared with hypoxia+negative-sequence control,HIF-2αsmall interfering ribonucleic acid transfection can lower the expression of HIF-2αand CD44 mRNA(P<0.05),increase the percentage of apoptotic cells(P<0.05),and resulted in a reduction of CD44+/CD24−po展开更多
目的鉴定乳腺癌干细胞系MCF-7中是否包含肿瘤干细胞相关的SP(side popu lation)亚群。方法制备MCF-7细胞悬液,经Hoechst33342和PI染色(其中一组同时加入拮抗剂维拉帕米作为对照),流式细胞仪分析SP亚群。另取贴壁生长的MCF-7细胞,用Hoech...目的鉴定乳腺癌干细胞系MCF-7中是否包含肿瘤干细胞相关的SP(side popu lation)亚群。方法制备MCF-7细胞悬液,经Hoechst33342和PI染色(其中一组同时加入拮抗剂维拉帕米作为对照),流式细胞仪分析SP亚群。另取贴壁生长的MCF-7细胞,用Hoechst33342和羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)原位染色,荧光显微镜下观察细胞染色情况。结果SP亚群占MCF-7细胞的4%,维拉帕米阻断后减少为0.5%。荧光显微镜下见Hoechst33342阳性细胞约占95%。结论人乳腺癌细胞系MCF-7中存在SP亚群,即提示乳腺癌干细胞的存在;维拉帕米可抑制染料外排而减少SP细胞的比例。展开更多
Cancer stem cells （CSCs）, a subpopulation of cancer cells with ability of initiating tumorigenesis, exist in many kinds of tumors including breast cancer. Cancer stem cells contribute to treatment resistance and rel...Cancer stem cells （CSCs）, a subpopulation of cancer cells with ability of initiating tumorigenesis, exist in many kinds of tumors including breast cancer. Cancer stem cells contribute to treatment resistance and relapse. Conventional treatments only kill differentiated cancer cells, but spare CSCs. Combining conventional treatments with therapeutic drugs targeting to CSCs will eradicate cancer cells more efficiently. Studying the molecular mechanisms of CSCs regulation is essential for developing new therapeutic strategies. Growing evidences showed CSCs are regulated by non-coding RNA （ncRNA） including microRNAs and long non-coding RNAs （IncRNAs）, and histone-modifiers, such as let- 7, miR-93, miR-100, HOTAIR, Bmi-1 and EZH2. Herein we review the roles of microRNAs, IncRNAs and histone- modifiers especially Polycomb family proteins in regulating breast cancer stem cells （BCSCs）.展开更多
This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells （BCSCs）. BCSCs were enriched by using serum-free me- dium and knocked out of N...This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells （BCSCs）. BCSCs were enriched by using serum-free me- dium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction （RT-PCR） and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and fl0w cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a prom- ising therapeutical approach for breast cancer.展开更多
MicroR NAs(miR NAs) are small non-coding RNAs generated by a two-step complex process and are post transcriptional negative regulators of their target m RNAs. Dysregulation of many of these miR NAs has been associated...MicroR NAs(miR NAs) are small non-coding RNAs generated by a two-step complex process and are post transcriptional negative regulators of their target m RNAs. Dysregulation of many of these miR NAs has been associated with tumorigenesis in various cancers including breast cancer. Aberrantly high expression of specific mi RNAs in breast cancer cells is demonstrated to be linked with inhibition of tumor suppressor genes and promote tumorigenesis. They are classified as oncogenic miR NAs. However, the tumor suppressor miR NAs are downregulated in breast cancer cells, since their major targets are oncogenic m RNAs. Understanding mechanism of action of specific miR NAs in breast cancer cells can be utilized to develop newer anti-cancer therapies. Recently, newer techniques are also developed to detect abundance of specific miR NA in the blood plasma samples and can be used in early diagnosis or prognosis in breast cancer. In this review article, we have discussed several mi RNAs dysregulated in breast cancer and their therapeutic potential.展开更多
Signaling within the tumor microenvironment has a critical role in cancer initiation and progression. Adipocytes, one of the major components of the breast microenvironment,have been shown to provide pro-tumorigenic s...Signaling within the tumor microenvironment has a critical role in cancer initiation and progression. Adipocytes, one of the major components of the breast microenvironment,have been shown to provide pro-tumorigenic signals that promote cancer cell proliferation and invasiveness in vitro and tumorigenicity in vivo. Adipocyte secreted factors such as leptin and interleukin-6(IL-6) have a paracrine effect on breast cancer cells. In adipocyte-adjacent breast cancer cells, the leptin and IL-6 signaling pathways activate janus kinase 2/signal transducer and activatorof transcription 5, promoting the epithelial-mesenchymal transition, and upregulating stemness regulators such as Notch, Wnt and the Sex determining region Y-box 2/octamer binding transcription factor 4/Nanog signaling axis. In this review we will summarize the major signaling pathways that regulate cancer stem cells in breast cancer and describe the effects that adipocyte secreted IL-6 and leptin have on breast cancer stem cell signaling. Finally we will introduce a new potential treatment paradigm of inhibiting the adipocyte-breast cancer cell signaling via targeting the IL-6 or leptin pathways.展开更多
Breast cancer remains a leading cause of morbidity and mortality in women mainly because of the propensity of primary breast tumors to metastasize. It is composed of heterogeneous cell populations with different biolo...Breast cancer remains a leading cause of morbidity and mortality in women mainly because of the propensity of primary breast tumors to metastasize. It is composed of heterogeneous cell populations with different biological properties. Breast cancer-initiating cells have been recently identified in breast carcinoma as CD44+/CD24-/low cells, which display stem cell like properties. In the present study, we have isolated breast cancer stem cells from non-metastasis tumor tissue, which is presently at passage 18 and designated as human Breast Cancer Mesenchymal Stem Cells (hBCMSCs) line. These cells showed spindle shaped morphology and formed mammos-pheres as well as pluripotency clones indicating their stem cell nature. Molecular marker study confirmed mesenchymal nature as well as pluripotency, plasticity and oncogenicity of these cells. The hBCMSCs cell line may likely contain a heterogeneous population of malignant cells. Interestingly, we also found that these cells exhibit BRCA 2 mutation, which was found in Indian population. Overall, this study revealed that hBCMSCs cell line may represent a suitable in vitro model to study the mechanism of breast cancer which further leads to an identification of molecular targets for future breast cancer targeted therapy.展开更多
背景:乳腺癌是女性最常见的恶性肿瘤之一,其发病率逐年上升且趋于年轻化,严重威胁女性生命健康,因此建立一个能够准确模拟体内肿瘤情况的理想乳腺癌模型显得尤为重要。类器官与体内来源的组织或器官高度相似,是一种新型的体外三维培养模...背景:乳腺癌是女性最常见的恶性肿瘤之一,其发病率逐年上升且趋于年轻化,严重威胁女性生命健康,因此建立一个能够准确模拟体内肿瘤情况的理想乳腺癌模型显得尤为重要。类器官与体内来源的组织或器官高度相似,是一种新型的体外三维培养模型,近年来在各种恶性肿瘤的研究中备受研究人员的青睐。目的:综述乳腺癌类器官的研究进展及应用,以期为乳腺癌的个性化治疗提供新的研究途径。方法:第一作者以“类器官、乳腺癌类器官、肿瘤类器官、乳腺球、3D培养”“organoid,breast cancer organoids,cancer organoids,mammosphere,three-dimensional culture”为检索词,检索1980年1月至2020年2月CNKI、万方、PubMed数据库相关文献,文献类型不限,排除与文章研究目的无关及重复性的文献,最终纳入66篇文献进行综述分析。结果与结论:文章简要介绍了类器官技术,回顾了构建乳腺癌类器官模型的过程,总结了乳腺癌类器官最新研究进展及应用。乳腺癌类器官在疾病建模、肿瘤发生机制、药物筛选等方面有广泛应用前景,为乳腺癌研究和治疗提供了可靠的模型,特别是为乳腺癌个性化治疗开辟了新的视野。展开更多
文摘BACKGROUND Breast cancer is a common malignant tumor that seriously threatens women’s health.Breast cancer stem cell(CSC)-like cell population may be the main factor for breast cancer metastasis.Therefore,targeted therapy for CSCs has great potential significance.Hypoxia-inducible factor is a transcription factor widely expressed in tumors.Studies have shown that down-regulation of the hypoxia signaling pathway inhibits tumor stem cell self-renewal and increases the sensitivity of stem cells to radiotherapy and chemotherapy mediated by hypoxiainducible factor-2α(HIF-2α).However,the specific mechanism remains unclear and further research is necessary.AIM To investigate the effect of HIF-2αdown-regulation on stem cell markers,microsphere formation,and apoptosis in breast cancer cell line MDA-MB-231 under hypoxia and its possible mechanism.METHODS Immunohistochemistry was used to detect the expression of HIF-2αand CD44 in triple-negative breast cancer(TNBC)and non-TNBC tissues.Double-labeling immunofluorescence was applied to detect the co-expression of HIF-2αand CD44 in MDA-MB-231 cells and MCF-7 cells.HIF-2αwas silenced by RNA interference,and the expression of CD44 and transfection efficiency were detected by real-time fluorescent quantitative PCR.Further,flow cytometry,TdT-mediated X-dUTP nick end labeling,and mammosphere formation assays were used to evaluate the effect of HIF-2αon CSCs and apoptosis.The possible mechanisms were analyzed by Western blot.RESULTS The results of immunohistochemistry showed that HIF-2αwas highly expressed in both TNBC and non-TNBC,while the expression of CD44 in different molecular types of breast cancer cells was different.In in vitro experiments,it was found that HIF-2αand CD44 were expressed almost in the same cell.Compared with hypoxia+negative-sequence control,HIF-2αsmall interfering ribonucleic acid transfection can lower the expression of HIF-2αand CD44 mRNA(P<0.05),increase the percentage of apoptotic cells(P<0.05),and resulted in a reduction of CD44+/CD24−po
文摘目的鉴定乳腺癌干细胞系MCF-7中是否包含肿瘤干细胞相关的SP(side popu lation)亚群。方法制备MCF-7细胞悬液,经Hoechst33342和PI染色(其中一组同时加入拮抗剂维拉帕米作为对照),流式细胞仪分析SP亚群。另取贴壁生长的MCF-7细胞,用Hoechst33342和羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)原位染色,荧光显微镜下观察细胞染色情况。结果SP亚群占MCF-7细胞的4%,维拉帕米阻断后减少为0.5%。荧光显微镜下见Hoechst33342阳性细胞约占95%。结论人乳腺癌细胞系MCF-7中存在SP亚群,即提示乳腺癌干细胞的存在;维拉帕米可抑制染料外排而减少SP细胞的比例。
文摘Cancer stem cells （CSCs）, a subpopulation of cancer cells with ability of initiating tumorigenesis, exist in many kinds of tumors including breast cancer. Cancer stem cells contribute to treatment resistance and relapse. Conventional treatments only kill differentiated cancer cells, but spare CSCs. Combining conventional treatments with therapeutic drugs targeting to CSCs will eradicate cancer cells more efficiently. Studying the molecular mechanisms of CSCs regulation is essential for developing new therapeutic strategies. Growing evidences showed CSCs are regulated by non-coding RNA （ncRNA） including microRNAs and long non-coding RNAs （IncRNAs）, and histone-modifiers, such as let- 7, miR-93, miR-100, HOTAIR, Bmi-1 and EZH2. Herein we review the roles of microRNAs, IncRNAs and histone- modifiers especially Polycomb family proteins in regulating breast cancer stem cells （BCSCs）.
基金supported by the National Nature Science Foundation of China(No.30972935)
文摘This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells （BCSCs）. BCSCs were enriched by using serum-free me- dium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction （RT-PCR） and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and fl0w cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a prom- ising therapeutical approach for breast cancer.
基金Supported by In part supported by the American Cancer Society Research Award,No.RSG-10-067-01-TBENIH grant to Chen H,No.1R01CA178386
文摘MicroR NAs(miR NAs) are small non-coding RNAs generated by a two-step complex process and are post transcriptional negative regulators of their target m RNAs. Dysregulation of many of these miR NAs has been associated with tumorigenesis in various cancers including breast cancer. Aberrantly high expression of specific mi RNAs in breast cancer cells is demonstrated to be linked with inhibition of tumor suppressor genes and promote tumorigenesis. They are classified as oncogenic miR NAs. However, the tumor suppressor miR NAs are downregulated in breast cancer cells, since their major targets are oncogenic m RNAs. Understanding mechanism of action of specific miR NAs in breast cancer cells can be utilized to develop newer anti-cancer therapies. Recently, newer techniques are also developed to detect abundance of specific miR NA in the blood plasma samples and can be used in early diagnosis or prognosis in breast cancer. In this review article, we have discussed several mi RNAs dysregulated in breast cancer and their therapeutic potential.
文摘Signaling within the tumor microenvironment has a critical role in cancer initiation and progression. Adipocytes, one of the major components of the breast microenvironment,have been shown to provide pro-tumorigenic signals that promote cancer cell proliferation and invasiveness in vitro and tumorigenicity in vivo. Adipocyte secreted factors such as leptin and interleukin-6(IL-6) have a paracrine effect on breast cancer cells. In adipocyte-adjacent breast cancer cells, the leptin and IL-6 signaling pathways activate janus kinase 2/signal transducer and activatorof transcription 5, promoting the epithelial-mesenchymal transition, and upregulating stemness regulators such as Notch, Wnt and the Sex determining region Y-box 2/octamer binding transcription factor 4/Nanog signaling axis. In this review we will summarize the major signaling pathways that regulate cancer stem cells in breast cancer and describe the effects that adipocyte secreted IL-6 and leptin have on breast cancer stem cell signaling. Finally we will introduce a new potential treatment paradigm of inhibiting the adipocyte-breast cancer cell signaling via targeting the IL-6 or leptin pathways.
文摘Breast cancer remains a leading cause of morbidity and mortality in women mainly because of the propensity of primary breast tumors to metastasize. It is composed of heterogeneous cell populations with different biological properties. Breast cancer-initiating cells have been recently identified in breast carcinoma as CD44+/CD24-/low cells, which display stem cell like properties. In the present study, we have isolated breast cancer stem cells from non-metastasis tumor tissue, which is presently at passage 18 and designated as human Breast Cancer Mesenchymal Stem Cells (hBCMSCs) line. These cells showed spindle shaped morphology and formed mammos-pheres as well as pluripotency clones indicating their stem cell nature. Molecular marker study confirmed mesenchymal nature as well as pluripotency, plasticity and oncogenicity of these cells. The hBCMSCs cell line may likely contain a heterogeneous population of malignant cells. Interestingly, we also found that these cells exhibit BRCA 2 mutation, which was found in Indian population. Overall, this study revealed that hBCMSCs cell line may represent a suitable in vitro model to study the mechanism of breast cancer which further leads to an identification of molecular targets for future breast cancer targeted therapy.
文摘背景:乳腺癌是女性最常见的恶性肿瘤之一,其发病率逐年上升且趋于年轻化,严重威胁女性生命健康,因此建立一个能够准确模拟体内肿瘤情况的理想乳腺癌模型显得尤为重要。类器官与体内来源的组织或器官高度相似,是一种新型的体外三维培养模型,近年来在各种恶性肿瘤的研究中备受研究人员的青睐。目的:综述乳腺癌类器官的研究进展及应用,以期为乳腺癌的个性化治疗提供新的研究途径。方法:第一作者以“类器官、乳腺癌类器官、肿瘤类器官、乳腺球、3D培养”“organoid,breast cancer organoids,cancer organoids,mammosphere,three-dimensional culture”为检索词,检索1980年1月至2020年2月CNKI、万方、PubMed数据库相关文献,文献类型不限,排除与文章研究目的无关及重复性的文献,最终纳入66篇文献进行综述分析。结果与结论:文章简要介绍了类器官技术,回顾了构建乳腺癌类器官模型的过程,总结了乳腺癌类器官最新研究进展及应用。乳腺癌类器官在疾病建模、肿瘤发生机制、药物筛选等方面有广泛应用前景,为乳腺癌研究和治疗提供了可靠的模型,特别是为乳腺癌个性化治疗开辟了新的视野。
