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miR-888 in MCF-7 side population Sphere Cells Directly Targets E-cadherin

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Journal of Genetics and Genomics 2014年第1期41卷 35-42页

作者： Shengjian Huang Ming Cai Yinghui Zheng Longhai Zhou Qiang Wang Liangbiao Chen Key Laboratory of Molecular and Developmental Biology Institute of Genetics and Developmental BiologyChinese Academy of Sciences The Department of Urology of PLA 309 Hospital

side population （SP） cells are a small subset of cells isolated from a cultured cancer cell line that exhibit characteristics similar to those of cancer stem cells （CSCs）, such as high metastatic and tumorigenic potential. The molecular mechanisms that give rise to the malignant properties of SP cells are not clear. We isolated SP cells from the MCF-7 breast cancer cell line and profiled microRNA （miRNA） expression patterns between SP cell-derived spheroids and non-SP cells. SP spheroids were found to possess 42 up-regulated miRNAs and 27 down-regulated ones （above 5-fold changes）. One of the up-regulated miRNAs, miR-888 computationally predicted to participate in the adherens junction （AJ） pathway, was investigated. Over-expression of miR-888 in MCF-7 cells reduced the mRNA levels of all four AJ pathway genes （E-cadherin, ACTG1, PTPRTand CDC42） that were selected for testing, whereas knocking down miR-888 reversed the trends. Western blot and flow cytometric quantitation of the membrane E-cadherin levels showed the same trend of change under these treatments. Luciferase reporter assay showed E-cadherin is a direct target of miR-888. As a potential role in intercellular adhesiveness and maintenance of malignant tissue architecture, the results indicate that miR-888 is a repressor of the AJ pathway in MCF-7 cells and that up-regulation of miR-888 contributes to aggressiveness in MCF-7 SP cells.

关键词： side population Cancer stem cell MicroRNA Adherens junction Cancer metastasis

Identification and Characterization of side population Cells in Human Lung Adenocarcinoma SPC-A1 Cells

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Chinese Journal of Cancer Research 2010年第3期22卷 211-217页

作者： Yan-liang Zhu Long-bang Chen Jing-hua Wang Xin-yi Xia Department of Oncology Department of Clinical Laboratory Centre Jinling HospitalMedical College of Nanjing University Nanjing 210002 China

Objective：There has been an increasing interest in recent years in the role of stem *** an extensive understanding of their biology,a major role for stem cells in the malignant process has been proposed and the existence of cancer stem cells（CSCs） has been confirmed in hematopoietic malignancies and solid organ malignancies including brain cancer,breast,prostate,colon,and pancreatic *** cancer is the leading cause of cancer mortality in most large cities of *** is possible that lung cancer contains cancer stem cells responsible for its *** aim of this study is to identify,characterize and enrich the CSC population that drives and maintains lung adenocarcinoma growth and ***：side population（SP） cell analysis and sorting were applied on human lung adenocarcinoma cell line and an attempt to further enrich them by preliminary serum-free culture before fluorescence activated cell sorting（FACS） was *** cell properties of SP cells were evaluated by their proliferative index,colony-forming efficiency,tumorigenic potential,bi-differentiation capacity and the expression of common stem cell surface ***：Lung cancer cells could grow in a serum-free Medium（SFM） as non-adherent spheres similar to neurospheres or *** proportion of SP cells in cell spheres was significantly higher than that in cells grown as *** cells had a greater proliferative index,a higher colony-forming efficiency and a greater ability to form tumor in *** cells were both CCA positive and SP-C positive while non-SP cells were only SP-C *** cytometric analysis of cell phenotype showed that SP cells expressed CD133 and CD44,the common cell surface markers of cancer stem cells,while non-SP cells only expressed ***：SP cells existed in human lung adenocarcinoma cell lines and they could be further enriched by preliminary serum-free culture before FACS *** cells possessed the properties of cancer st

关键词： Cancer stem cells side population SPC-A1 cell line

Effect of Bcl-2 and Bax on survival of side population cells from hepatocellular carcinoma cells

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World Journal of Gastroenterology 2007年第45期13卷 6053-6059页

作者： Jing Fan Ren Li Rui Zhang Hai-Liang Liu Ning Zhang Fu-Qing Zhang Ke-Feng Dou Department of Hepatobiliary Surgery Xijing Hospital the Fourth Military Medical University Xi'an 710032 Shaanxi Province China Department of Biochemistry and Molecular Biology the Fourth Military Medical University Xi'an 710032 Shaanxi Province China

AIM： To understand the role and significance of side population （SP） cells from hepatocellular carcinoma （HCC） in hepatocarcinogenesis, development, relapse and metastasis, we simulated the denutrition conditions that cancer cells experience in clinical therapy, observed the different anti-apoptosis ability of SP cells and non-SP cells under such conditions, and established the possible effects of P53, Bcl-2 and Bax on survival of SP cells. METHODS： We used flow cytometry to analyze and sort the SP and non-SP cells in established HCC lines MHCC97 and hHCC. We evaluated cell proliferation by methyl thiazolyl tetrazolium （MTT） assay and investigated the expression of p53, bcl-2 and bax genes during denutrition, by RT-PCR and immunofluorescence staining. RESULTS： The percentage of SP cells in the two established HCC lines was 0.25% and 0.5%, respectively. SP cells had greater anti-apoptosis and proliferation ability than non-SP cells. Expression of Bcl-2 and Bax in SP and non-SP cells differed during denutrition. The former was up-regulated in SP cells, and the latter was up-regulated in non-SP cells. CONCLUSION： It may be that different upstream molecules acted and led to different expression levels of Bcl-2 and Bax in these two cell lines. There was a direct relationship between up-regulation of Bcl-2 and down-regulation of Bax and higher anti-apoptosis ability in SP cells. It may be that the existence and activity of SP cells are partly responsible for some of the clinical phenomena which are seen in HCC, such as relapse or metastasis. Further research on SP cells may have potential applications in the field of anticancer therapy.

关键词： side population Hepatocellular carcinoma Bax Bcl-2 Apoptosis

Distinct side population Cell Subtypes Have Different Stemness Levels in Human Ovarian Cancer Cells

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Current Medical Science 2021年第1期41卷 127-132页

作者： Yan-jie WENG Xiao-xiao ZHANG Xue WU Li-li GUO Chang-yu WANG Department of Obstetrics and Gynecology Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan430030China

The stemness of different side population(SP)cell subtypes in ovarian cancer cells was studied,and the heterogeneity of ovarian cancer stem cells was *** cisplatin-resistant human serous ovarian cancer cell line C13 was stained with the bisbenzimide Hoechst 33342.A flow cytometry-based fluorescence-activated sorting method was used to obtain lower-SP(LSP)cells,upper-SP(USP)cells,and non-SP cells(NSP)based on their sensitivity to the staining time and Hoechst dye *** sphere-forming capability,expression levels of stem cell markers,resistance to high concentrations of cisplatin,and subcutaneous tumorigenicity in NOD/SCID mice of the different cell subtypes were *** C13 cells contained SP cells with stemness characteristics,and the LSP cell subtype expressed higher levels of stem cell markers,had higher in vitro sphere-forming capability,higher cisplatin resistance and higher in vivo subcutaneous tumorigenesis than USP cells(P<0.05).NSP cells had no *** conclusion,different subtypes of ovarian cancer SP cells have different stemness levels,and ovarian cancer stem cells may be heterogeneous.

关键词： ovarian cancer cancer stem cells heterogeneity side population

Heterogeneity beyond tumor heterogeneity—SULF2 involvement in Wnt/β-catenin signaling activation in a heterogeneous side population of liver cancer cells

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BIOCELL 2023年第9期47卷 2037-2049页

作者： DONGYE YANG DONGDONG GUO YUNMEI PENG DONGMENG LIU YANQIU FU FEN SUN LISHI ZHOU JIAQI GUO LAIQING HUANG Division of Gastroenterology&Hepatology The University of Hong Kong-Shenzhen HospitalShenzhen518053China Shenzhen Key Laboratory of Gene and Antibody Therapy Center for Biotechnology and BiomedicineState Key Laboratory of Health Sciences and TechnologyState Key Laboratory of Chemical OncogenomicsPrecision Medicine and Healthcare Research CenterTsinghua-Berkeley Shenzhen Institute(TBSI)Shenzhen International Graduate SchoolTsinghua UniversityShenzhen518055China Department of Chemistry Tsinghua UniversityBeijing100084China

Introduction:Sulfatase 2(SULF2),an endogenous extracellular sulfatase,can remove 6-O-sulfate groups of glucosamine residues from heparan sulfate(HS)chains to modulate the Wnt/β-catenin signaling pathway,which plays an important role in both liver carcinogenesis and *** population(SP)cells are widely identified as stem-like cancer cells and are closely related to carcinoma metastasis,recurrence,and poor patient ***,the roles of SULF2 in SP cells of hepatomas are unclear,and the underlying mechanism is ***:This study aimed to compare the heterogeneity between SP cells and non-side population(NSP)cells derived from three different liver cancer cell lines and to elucidate the involvement of the SULF2-Wnt/β-catenin axis in liver cancer stem cells(CSCs)and its impact on the processes of carcinogenesis and ***:In this work,three different liver cancer SP cells(HepG2,Huh7,and PRC/PRL/5)were sorted by flow *** also examined the migration and invasion behaviors of SP and NSP *** determine if this high tumorigenic potential of SP cells is correlated to SULF2,qPCR,western blotting,and immunofluorescence analysis were *** also performed nude mouse xenograft experiments for in vivo ***:The results from the in vitro colony formation assay showed that SP cells exhibited a 2-fold higher colony formation efficiency compared to their NSP *** SP cells exhibited significantly higher potentials in terms of their migratory capacity and invasive ability compared to NSP *** found that higher expression of SULF2 in SP cells was associated with greater capabilities for clonogenicity,migration,and *** was also linked to higher activation of the Wnt/β-catenin signaling pathway via stimulation of key downstream factors,particularlyβ-catenin,c-Myc,and cyclin ***,a positive correlation between the upregulated SULF2 expression and tumorigenesis in the in vivo nude mous

关键词： SULF2 Sulfatase2 Liver cancer side population Wnt/β-catenin pathway

Identification and analysis of a minority fraction in a U87 cell line Do side-population cells represent bona fide stem cell-like cancer cells?

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Neural Regeneration Research 2009年第11期4卷 805-813页

作者： Changwei Gu Qingfang Sun Jiang Zhu Liuguan Bian Weiguo Zhao Jiankang Shen HengGao Dongxia Feng l lnternational Cooperation of Neuro-centre in Jiangyin People's HospitaI Jiangyin 214400 Jiangsu Province China Department of Neurosurgery Ruijin Hspitaal School of Medicine Shanghai Jiao Tong University Shanghai 200025 China Shanghai Institute of Hematology Ruijin Hospital Shanghai 200025 China

BACKGROUND： Overwhelming evidence suggests that tumor bulks are comprised of differentiated tumor cells and cancer stem cells （CSCs）. The stem cell-like side-population （SP） cells account for a minor fraction of the total tumor cells, yet are apparently the cells capable of tumor initiation, growth, maintenance, and recurrence. OBJECTIVE： To identify potential stem cell-like cancer cells in a U87 human brain glioma cell line on the basis of dye efflux, clone formation, and multi-drug resistance capacity. DESIGN, TIME AND SETTING： The cellular and molecular biology experiment was performed at the Laboratory of Shanghai Institute of Hematology and Laboratory of Shanghai Institute of Endocrinology in Ruijin Hospital; in vivo contrast observational animal trial was performed at Experimental Animal Center, School of Medicine, Shanghai Jiao Tong University from June 2007 to May 2008. MATERIALS： The U87 cell line was provided by the Shanghai Institute of Cancer Research, Chinese Academy of Science; DMEM/F12 （1 ： 1） and fetal bovine serum were purchased from Gibco Invitrogen, USA; human recombinant basic fibroblast growth factors were purchased from BD Bioscience, USA; Hoechst 33342, Verapamil, and methyl thiazolyl tetrazolium were purchased from Sigma, USA; phycoerythrin-labeled anti-human-CD133 was purchased from Milteny Biotec, Germany; SYBR PrimeScriptTM RT-PCR kit was purchased from TaKaRa Biotechnology, Dalian, China. METHODS： Monolayer cultured cells were harvested by 0.25% Trypsin-EDTA and suspended at a 1 ×10^6/mL dilution in PBS containing 2% FBS, and were stained with Hoechst 33342 dye, either alone or in combination with Verapamil. Following fluorescence-activated cell sorting, SP and non-SP subsets were cultivated with serum-containing （DMEM plus 10% fetal bovine serum） or serum-free culture medium [DMEM/F12 （1： 1）＋ 1× B27 supplement ＋ 10 ng/mL basic fibroblast growth factors ＋ 1× L-glutamine] to determine growth characteristics in vitro. Finally, single free

关键词： cancer stem cells tumor-initiating cells side population glioma CD133

Endometrial stem/progenitor cells: Properties, origins, and functions

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Genes & Diseases 2023年第3期10卷 931-947页

作者： In-Sun Hong Department of Health Sciences and Technology GAIHSTGachon UniversityIncheon 21999Republic of Korea Department of Molecular Medicine School of MedicineGachon UniversityIncheon 21999Republic of Korea

The endometrium is the inner mucosal lining of the uterus that undergoes extensive cyclic growth, regeneration, differentiation, and shedding throughout the menstrual cycle in response to steroid hormones. It repeatedly undergoes approximately 450 cycles of degeneration and regeneration in a woman's lifetime. Endometrial abnormalities can be associated with repeated embryo implantation failure, recurrent spontaneous abortion, and other physiological features responsible for female infertility. This significant regenerative capacity may occur as a result of tissue-resident stem cell populations within the endometrium. Indeed, the existence of endometrial stem cells was only observed in humans and rodents through several isolation and characterization methods in the last few years. Although endometrial stem cells share various biological characteristics with other types of mesenchymal stem cells, they also show some differences in phenotype, self-renewal, and multilineage differentiation potential. Extensive studies over many years on endometrial stem cells will provide new insights into the physiology and mechanisms underlying various gynaecological diseases related to endometrial abnormalities such as female infertility, endometriosis, and endometrial cancer. Here we summarized recent studies about cellular origins and biological characteristics of endometrial stem cells. We also reviewed various recent studies to improve our understanding of their physiological roles. Many preclinical studies on their potential therapeutic applications to various endometrial diseases that could lead to reproductive dysfunction were also reviewed.

关键词： CD146 LGR5 0CT-4 side population S0X9 SSEA-1 SUSD2 Wnt/β-catenin

Sox2 enhances the tumorigenicity and chemoresistance of cancer stem-like cells derived from gastric cancer

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The Journal of Biomedical Research 2012年第5期26卷 336-345页

作者： Tian Tian Yajie Zhang Shouyu Wang Jianwei Zhou Shan Xu Department of Cell Biology Cancer Center Nanjing Medical University Nanjing Jiangsu 210029 China Department of Molecular Cell Biology & Toxicology School of Public Health Nanjing Medical University Nanjing Jiangsu 210029 China

Gastric cancer stem-like cells（GCSCs） have been identified to possess the ability of self-renewal and tumor ***,the mechanisms involved remain largely ***,we isolated and characterized the GCSCs by side population（SP） sorting procedure and cultured sphere cells（SC） from human gastric cancer cell lines SGC-7901,BGC-823,MGC-803,HGC-27 and *** sorting and culture assay revealed that SP cells proliferated in an asymmetric division *** addition,SP cells exhibited a higher potential of spheroid colony formation and greater drug resistance than non-SP cells（NSP）.Moreover,the SC were found with enhanced capabilities of drug resistance in vitro and tumorigenicity in ***2 mRNA and protein was highly and significantly overex-pressed in the SP cells and ***,downregulation of Sox2 with siRNA obviously reduced spheroid colony formation and doxorubicin efflux,as well as increased apoptosis rate in sphere cells in vitro and suppressed tumori-genicity in *** results suggest that both SP cells and cultured SC enrich with GCSCs and that Sox2 plays a pivotal role in sustaining stem cell properties and might be a potential target for gastric cancer therapy.

关键词： side population gastric cancer stem-like cells CD44 Sox2 chemoresistance

Comparative study on the stem cell phenotypes of C6 cells under different culture conditions

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Chinese Medical Journal 2011年第19期124卷 3118-3126页

作者： ZHANG Suo-jun YE Fei XIE Rui-fan HU Feng WANG Bao-feng WAN Feng GUO Dong-sheng LEI Ting Department of Neurosurgery and Neuro-Oncology Molecular Biology Laboratory Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei 430030 China

Background Glioma stem cell （GSC） hypothesis posits that a subpopulation of cells within gliomas have true clonogenic and tumorigenic potential. Significantly, a more controversial correlate to GSC is that cells in different culture conditions might display distinct stem cell properties. Considering these possibilities, we applied an approach comparing stem cell characteristics of C6 glioma cells under different culture *** C6 cells were cultured under three different growth conditions, i.e., adherent growth in conventional 10% serum medium, non-adherent spheres growth in serum-free medium, as well as adherent growth on laminin-coated flask in serum-free medium. Growth characteristics were detected contrastively through neurosphere formation assay and cell cycle analysis. Markers were determined by immunofluorescence, relative-quantitative reverse transcription （RT）-PCR,Western blotting and flow cytometry. side population cells were analyzed via flow cytometry. Tumor models were detected by magnetic resonance imaging and hematoxylin ＆ eosin staining. Data analyses were performed with SPSS software （17.0）.Results C6 cells （C6-Adh, C6-SC-Sph and C6-SC-Adh） showed distinctive growth patterns and proliferation *** to suspending C6-SC-Sph, adherent C6-Adh and C6-SC-Adh displayed higher growth ratio. C6-SC-Sph and C6-SC-Adh showed enhanced capability of neurosphere formation and self-renewal. High side population ratio was detected in C6-SC-Sph and C6-SC-Adh. CD133 was not detected in all three kinds of cells. Conversely, Nestin and β-Ⅲ-tubulin were demonstrated positive, nonetheless with no statistical significance （P 〉0.05）. Interestingly, lower expression of glial fibrillary acidic protein was demonstrated in C6-SC-Sph and C6-SC-Adh. C6-Adh, C6-SC-Sph and C6-SC-Adh were all displayed in situ oncogenicity, while statistical difference of survival time was not *** C6 glioma cell line is endowed with some GSC phenotypes that

关键词： C6 glioma stem cell neurosphere CD133 side population laminin