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Cancer Stem-Like cells directly participate in vasculogenic mimicry channels in triple-negative breast cancer

Cancer Biology & Medicine 2019年第2期16卷 299-311页

作者： Huizhi Sun Nan Yao Siqi Cheng Linqi Li Shiqi Liu Zhao Yang Guanjie Shang Danfang Zhang Zhi Yao Department of Pathology Tianjin Medical University Tianjin 300070 China Department of Pathology General Hospital of Tianjin Medical University Tianjin 300070 China Department of Immunology Tianjin Medical University Tianjin 300070 China

Objective: Vasculogenic mimicry(VM) channels that are lined by tumor cells are a functional blood supply in malignant tumors.However, the role of VM-initiating cells remains poorly understood. Cancer Stem-Like cells(CSCs) are positively correlated with VM. In this study, triple-negative breast cancer(TNBC) enriched with CSCs was used to investigate the relationship between VM and CSCs.Methods: The expression of several CSC markers was detected by immunohistochemistry in 100 human breast cancer samples.The clinical significance of CSC markers and the relationship between VM, CSCs, breast cancer subtypes, and VM-associated proteins were analyzed. CD133+ and ALDH+ human and mouse TNBC cells were isolated by FACS to examine the ability of VM formation and the spatial relationship between VM and CSCs.Results: CSCs were associated with TNBC subtype and VM in human invasive breast cancer. CSCs in TNBC MDA-MB-231 cells formed more VM channels and expressed more molecules promoting VM than the non-TNBC MCF-7 cells in vitro. MDA-MB-231 cells that encircled VM channels on Matrigel expressed CD133. Moreover, CSCs were located near VM channels in the 3D reconstructed blood supply system in human TNBC grafts. The CD133+ and ALDH+ cells isolated from TA2 mouse breast cancer formed more VM channels in vivo.Conclusions: CSCs line VM channels directly. Additionally, CSCs provide more VM-related molecules to synergize VM formation. The signaling pathways that control CSC differentiation may also be potential treatment targets for TNBC.

关键词： Vasculogenic mimicry triple-negative breast cancer cancer Stem-Like cells ALDH1 CD 133

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Hallmarks in colorectal cancer:Angiogenesis and cancer Stem-Like cells

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World Journal of Gastroenterology 2014年第15期20卷 4189-4196页

作者： Muriel Mathonnet Aurelie Perraud Niki Christou Hussein Akil Carole Melin Serge Battu Marie-Odile Jauberteau Yves Denizot Department of Digestive Surgery Limoges University Hospital87042 LimogesFrance Homéostasie Cellulaire at Pathologies Faculty of MedicineUniversity of Limoges87025 LimogesFrance Institut 145 GEIST UMR CNRS 7276Faculty of MedicineUniversity of Limoges87025 LimogesFrance

Carcinogenesis is a multistep process that requires the accumulation of various genetic and epigenetic aberrations to drive the progressive malignant transformation of normal human cells.Two major hallmarks of carcinogenesis that have been described are angiogenesis and the stem cell characteristic of limitless replicative potential.These properties have been targeted over the past decade in the development of therapeutic treatments for colorectal cancer(CRC),one of the most commonly diagnosed and lethal cancers worldwide.The treatment of solid tumor cancers such as CRC has been challenging due to the heterogeneity of the tumor itself and the chemoresistance of the malignant cells.Furthermore,the same microenvironment that maintains the pool of intestinal stem cells that contribute to the continuous renewal of the intestinal epithelia also provides the necessary conditions for proliferative growth of cancer Stem-Like cells.These cancer Stem-Like cells are responsible for the resistance to therapy and cancer recurrence,though they represent less than 2.5%of the tumor mass.The stromal environment surrounding the tumor cells,referred to as the tumor niche,also supports angiogenesis,which supplies the oxygen and nutrients needed for tumor development.Anti-angiogenic therapy,such as with bevacizumab,a monoclonal antibody against vascular-endothelial growth factor,significantly prolongs the survival of metastatic CRC patients.However,such treatments are not completely curative,and a large proportion of patient tumors retain chemoresistance or show recurrence.This article reviews the current knowledge regarding the molecular phenotype of CRC cancer cells,as well as discusses the mechanisms contributing to their maintenance.Future personalized therapeutic approaches that are based on the interaction of the carcinogenic hallmarks,namely angiogenic and proliferative attributes,could improve survival and decrease adverse effects induced by unnecessary chemotherapy.

关键词： Colon cancer stem cell Cancer Stem-Like cell Tumor

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Xiaotan Sanjie decoction attenuates tumor angiogenesis by manipulating Notch-1-regulated proliferation of gastric cancer Stem-Like cells

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World Journal of Gastroenterology 2014年第36期20卷 13105-13118页

作者： Bing Yan Long Liu Ying Zhao Li-Juan Xiu Da-Zhi Sun Xuan Liu Ye Lu Jun Shi Yin-Cheng Zhang Yong-Jin Li Xiao-Wei Wang Yu-Qi Zhou Shou-Han Feng Can Lv Pin-Kang Wei Zhi-Feng Qin Department of Traditional Chinese Medicine Changzheng Hospital the Second Military Medical University Department of Traditional Chinese Medicine Changhai Hospital the Second Military Medical University

AIM: To determine the underlying mechanisms of action and influence of Xiaotan Sanjie(XTSJ) decoction on gastric cancer Stem-Like cells(GCSCs). METHODS: The gastric cancer cell line MKN-45 line was selected and sorted by FACS using the cancer stem cell marker CD44; the stemness of these cells was checked in our previous study. In an in vitro study, the expression of Notch-1, Hes1, Vascular endothelial growth factor(VEGF), and Ki-67 in both CD44-positive gastric cancer Stem-Like cells(GCSCs) and CD44-negative cells was measured by Western blot. The effect of XTSJ serum on cell viability and on the above markers was measured by MTT assay and Western blot, respectively. In an in vivo study, the ability to induce angiogenesis and maintenance of GCSCs in CD44-positive-MKN-45- and CD44-negative-engrafted mice were detected by immunohistochemical staining using markers for CD34 and CD44, respectively. The role of XTSJ decoction in regulating the expression of Notch-1, Hes1, VEGF and Ki-67 was measured by Western blot and real-time polymerase chain reaction.RESULTS: CD44+ GCSCs showed more cell proliferation and VEGF secretion than CD44-negative cells in vitro, which were accompanied by the high expression of Notch-1 and Hes1 and positively associated with tumor growth(GCSCs vs CD44-negative cells, 2.72 ± 0.25 vs 1.46 ± 0.16, P < 0.05) and microvessel density(MVD)(GCSCs vs CD44-negative cells, 8.15 ± 0.42 vs 3.83 ± 0.49, P < 0.001) in vivo. XTSJ decoction inhibited the viability of both cell types in a dose-dependent manner in vitro. Specifically, a significant difference in the medium-(82.87% ± 6.53%) and high-dose XTSJ groups(77.43% ± 7.34%) was detected at 24 h in the CD44+ GCSCs group compared with the saline group(95.42% ± 5.76%) and the low-dose XTSJ group(90.74% ± 6.57%)(P < 0.05). However, the efficacy of XTSJ decoction was reduced in the CD44- groups; significant differences were only detected in the high-dose XTSJ group at 48 h(78.57% ± 6.94%) and 72 h(72.12% ± 7.68%) when compared with the other CD44- groups(P

关键词： Gastric cancer Stem-Like cells Xiaotan Sanjie deco

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MiR-210 expression reverses radioresistance of Stem-Like cells of oesophageal squamous cell carcinoma

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世界临床肿瘤学杂志：英文版 2014年第5期5卷 1068-1077页

作者： Xin Chen Jia Guo Ru-Xing Xi Yu-Wei Chang Fei-Yang Pan Xiao-Zhi Zhang Radiation Department First Affiliated Hospital of Medical School of Xi'an Jiaotong University School of Medicine Queen's University Kingston K7L3N6 Ontario Canada

AIM: To investigate the expression of miR-210 and the role it plays in the cell cycle to regulate radioresistance in oesophageal squamous cell carcinoma(ESCC). METHODS: Mi R-210 expression was evaluated in 37 pairs of ESCC tissues and matched para-tumorous normal oesophageal tissues from surgical patients who had not received neoadjuvant therapy, and in the cells of two novel radioresistant cell lines, TE-1R and Eca-109 R, using quantitative reverse transcription-polymerase chain reaction(q RT-PCR). The transient up-regulation of mi R-210 expression in TE-1R and Eca-109 R cells was studied using liposomes and was confirmed using qR T-PCR. The rate of cell survival after a series of radio-treatment doses was evaluated using the cloneformation assay. Flow cytometry was used to detect the changes to the cell cycle patterns due to radiation treatment. RT-PCR and Western blot were used to detect the expression of ataxia telangiectasia mutated(ATM) and DNA dependent protein kinase(DNA-PKcs) after irradiation, and the cell sphere formation assay was used to evaluate the proliferative ability of the cancer Stem-Like cells.RESULTS: The level of mi R-210 expression was significantly decreased, by 21.3% to 97.2%, with the average being 39.2% ± 16.1%, in the ESCC tissues of most patients(81.1%, 30 of 37 vs patients with high mi R-210 expression, P 0.05). Early local recurrences(< 18 mo, n = 19) after radiotherapy were significantly related with low miR-210 expression(n = 13, P < 0.05). The level of mi R-210 was decreased by approximately 73%(vs TE-1, 0.27 ± 0.10, P < 0.01) in the established radioresistant TE-IR cell line and by 52%(vs Eca-109, 0.48 ± 0.17, P < 0.05) in the corresponding Eca-109 R line. Transient transfection with a mi R-210 precursor increased the level of mi R-210 expression, leading to a significant increase in cel

关键词： MiR-210 Oesophageal squamous cell carcinoma Radiation resistance Cell cycle arrest Stem-Like cells

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Carbon-Ion Beams Efficiently Induce Cell Killing in X-Ray Resistant Human Squamous Tongue Cancer Cells

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International Journal of Medical Physics, Clinical Engineering and Radiation Oncology 2014年第3期3卷 133-142页

作者： Akihisa Takahashi Hongyu Ma Akiko Nakagawa Yukari Yoshida Tatsuaki Kanai Tatsuya Ohno Yoshikazu Kuwahara Manabu Fukumoto Takashi Nakano Advanced Scientific Research Leaders Development Unit Gunma University Gunma Japan Department of Pathology IDAC Tohoku University Sendai Japan Department of Radiation Oncology Gunma University Graduate School of Medicine Gunma Japan Gunma University Heavy Ion Medical Center Gunma Japan

In order to see whether carbon ion (C-ion) beams have a biological advantage over X-rays, studies were designed to examine the effects of C-ion beams on radiosensitivity in X-ray resistant cells. Clinically relevant X-ray resistant SAS-R cells derived from human tongue cancer SAS cells were used. The cells were exposed to X-rays or Spread-Out Bragg peak (SOBP) beam C-ions. Cell survival was measured using a modified high-density survival assay. Cell survival signaling and cell death signaling were analyzed using flow cytometry. The cells were labeled with putative cancer stem cell markers such as CD44 and CD326. SAS-R cells were 1.6 times more radioresistant than SAS cells after exposure to X-rays. Cell survival was similar in each cell line after exposure to C-ion beams. SAS-R cells displayed enhanced cell survival signaling when compared to SAS cells under normal conditions. On the other hand, the phosphorylation of AKT-related proteins decreased and polycaspase activities were enhanced when cells were irradiated with C-ion beams in both cell lines. More CD44 and CD326 positive cells were seen in SAS-R cells than in SAS cells. Moreover, the marker positive cell numbers significantly decreased after exposure to C-ion beams when compared to X-rays at iso-survival doses in SAS-R cells. C-ion beams efficiently induced cell killing in X-ray resistant cells which displayed activated cell survival signaling and contained more numerous cancer Stem-Like cells.

关键词： Carbon-Ion Beams X-Ray Resistant Cells Cell Survival and Death Signaling Cancer Stem-Like Cells

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Discovery and characterization of novel small-molecule inhibitors targeting nicotinamide phosphoribosyltransferase

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中国药理学通报 2015年第B11期31卷 197-197页

作者： ZAHNGSai-long XUTian-ying MIAOChao-yu DepartmentofPharmacology SecondMilitaryMedicalUniversityShanghai200433

Aim Nicotinamide phosphoribosyltransferase （NAMPT） plays an important role in cardiocerebro-vascu- lar physiopathological process. It is also a promising anticancer target. It is highly desirable to discover novel NAMPT inhibitors as anticancer drug candidates and understand their action mode. Methods We carried out a high throughput screening system on a chemical library of 24434 small-molecules. Anti-proliferative activity were further studied on active compounds. Isothermal titration calorimetry and cellular thermal shift assay were used to confirm the target specificity. Molecular modeling and site-directed mutagenesis studies were taken to investigate the binding mode of NAMPT inhibitor. Results Using high throughput screening system targeting NAMPT, we ob- tained a potent NAMPT inhibitor MS0 （China Patent ZL201110447488.9 ） with excellent in vitro activity （IC50 = 9.87 ± 1.15 nmol · L^-1 ） and anti-proliferative activity against multiple human cancer cell lines including Stem-Like cancer cells. Structure-activity relationship studies yielded several highly effective analogues. These inhibitors spe- cifically bound NAMPT, rather than downstream NMNAT. We provided the first chemical case using cellular ther- mal shift assay to explain the difference between in vitro and cellular activity; MS7 showed best in vitro activity （ IC50 = 0.93 ± 0.29 nmol · L^-1 ） but worst cellular activity due to poor target engagement in living cells. Site-di- rected mutagenesis studies identified important residues for NAMPT catalytic activity and inhibitor binding. Con- clusions The present study provides a class of novel NAMPT inhibitors for future development of anticancer a- gents. Our findings also contribute to deep understanding the action mode of NAMPT inhibitors and NAMPT basic research in cardiocerebro-vascular system.

关键词： NAMPT inhibitor high throughput screening cancer Stem-Like cancer cells structure activity rela-tionship binding mode

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