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miR-612调控FOXP1影响白血病hl-60细胞的转移活性

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生物技术 2024年

作者：胡灵丹杜立坤刘影杜志强崔菁衡水市第二人民医院病理科衡水市第二人民医院血液科衡水市第二人民医院分子实验室

[目的]探究miR-612与FOXP1对急性髓系白血病(AML)细胞活性的影响。[方法]通过免疫组化实验分析正常组和AML组的骨髓样本中的FOXP1表达水平。将人急性髓系白血病细胞hl-60细胞分为4组:miR NC组、miR-612 mimic组、si NC组和siFOXP1组。通... 详细信息

[目的]探究miR-612与FOXP1对急性髓系白血病(AML)细胞活性的影响。[方法]通过免疫组化实验分析正常组和AML组的骨髓样本中的FOXP1表达水平。将人急性髓系白血病细胞hl-60细胞分为4组:miR NC组、miR-612 mimic组、si NC组和siFOXP1组。通过MTT实验检测细胞生长能力,实时荧光定量PCR实验检测miR-612和FOXP1表达水平,Transwell实验分析细胞迁移能力,蛋白免疫印迹实验分析FOXP1表达水平,流式细胞术分析细胞凋亡率。[结果]与正常组骨髓样本相比,AML组骨髓样本miR-612表达水平降低(Phl-60细胞的增殖能力和转移能力显著降低,凋亡率增加(Phl-60细胞的增殖能力和转移能力显著降低,凋亡率增加(Phl-60细胞的增殖能力和转移能力,并能促进hl-60细胞凋亡。此外,miR-612对hl-60细胞的这一作用与靶向抑制FOXP1表达有关。

关键词： miR-612 急性髓系白血病 FOXP1 转移侵袭 hl-60 凋亡增殖

全反式维甲酸诱导分化的hl-60在佛波酯刺激下产生中性粒细胞胞外诱捕网的特性

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生物医学工程学杂志 2022年第5期39卷 909-918页

作者：刘望方金花洪天添黄嘉祺赵柏松方颖吴建华林蒋国华南理工大学生物科学与工程学院广州510006 广东省人民医院广东省医学科学院医学研究部广州510080 广州市妇女儿童医疗中心麻醉科广州510623 广东省人民医院广东省医学科学院急诊科广州510080

中性粒细胞释放的胞外诱捕网(NETs)是一把双刃剑,了解NETs的形成机制对疾病治疗具有重要意义。然而,寿命短、个体差异大以及无法进行基因编辑使得中性粒细胞难以被用来破译NETs的形成。因此,寻找模式细胞替代中性粒细胞来研究NETs的形... 详细信息

中性粒细胞释放的胞外诱捕网(NETs)是一把双刃剑,了解NETs的形成机制对疾病治疗具有重要意义。然而,寿命短、个体差异大以及无法进行基因编辑使得中性粒细胞难以被用来破译NETs的形成。因此,寻找模式细胞替代中性粒细胞来研究NETs的形成机制是非常有必要的。在本研究中,我们使用不同浓度(0、0.1、1和10μmol/L)的全反式维甲酸(ATRA)对hl-60细胞进行不同天数的分化(1、3、5和7天)。通过检测细胞活力和细胞核形态,我们证实hl-60细胞经ATRA处理5天后可以分化为中性粒细胞样细胞(dhl-60)。使用免疫荧光染色检测NETs的形成,结果表明1μmol/L的ATRA分化5天得到的dhl-60细胞在佛波酯(PMA)刺激下产生的NETs与中性粒细胞产生的相当,且都不伴随组蛋白H3瓜氨酸化。此外,dhl-60细胞形成的NETs依赖于NADPH而不依赖于PAD4,这与中性粒细胞一致。这些结果表明,经1μmol/L ATRA分化5天的dhl-60细胞可作为中性粒细胞的模式细胞,用于NETs形成机制的研究。

关键词：中性粒细胞胞外诱捕网 hl-60 全反式维甲酸分化

Purification and Characterization of DNA Methylase From hl-60 Cells

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Science China Chemistry 1994年第4期37卷 430-436页

作者：何忠效王顺泰姚知行王秀奇 Department of Biology Beijing Normal University Beijing 100875 PRC State Key Laboratory of Macrobiomolecule Beijing 100101 PRC

A solid leukemia sarcoma has been successfully developed after subcutaneous inoculation of the cultured human promyelocytic leukemia cells (hl-60 cells) into unde mice. The solid leukemia sarcoma is a more plantiful source than the cultured cells for enzymatic study and its growing environment is closer to that of the human body than the cultured cells.We establish an efficient procedure of purifying hl-60 cells DNA methylase which includes: disruption of hl-60 cells by homogenization and sonication, removing the cell fragments and cellular particles by centrifuge and ultracentrifuge (105.000 g); removing endogenous DNA by streptomycin sulfate, salting out by (NH4)2SO4, ion exchange chromatography on DEAE-cellulose (DE-52), gel filtration over Sephadex G-100 column.The DNA methylase from hl-60 cells has been purified 204 fold by this procedure. The purified enzyme shows a single-band on PG-PAGE. A 479-kD molecular weight of this enzyme is measured by PG-PAGE. The enzyme properties of hl-60 DNA

关键词： hl-60 cells DNA methylase chromatography PG-PAGE.

A new 2-aminosteroid induces cellular differentiation and upregulates the expression of MafB and Egr-1 genes respectively in hl-60 and K562 leukemia cells

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Chinese Medical Journal 2005年第2期118卷 91-99页

作者： HEQun LIQiong YUANLin-bo HEJun ResearchlaboratoryofBloodPhysiology DepartmentofPhysiologyXiangyaSchoolofMedicineCentralSouthUniversityChangsha410078China

Background In previous work, we suggested that some 2aminosteroids inhibited proliferation and induced differentiation of both human and murine leukemia cells Here, we reported the actions of another new 2aminosteroid designated as H89712 on human leukemia cells Methods Cell colony counting and MTT assay were used to determine proliferation Cell morphology, histochemical staining, UV detection and cytometry were used to determine differentiation RTPCR was used to detect gene expression. Standard statistical method was used to analyze dataResults H89712 inhibited proliferation of hl60 leukemia cells and the inhibition percentage in MTT assay was 18% at the dose of 108 mol/L and 65% at the dose of 105 mol/L, respectively The inhibition for hl60 in colony assay was 23% at the dose of 108 mol/L and 96% at the dose of 105 mol/L, respectively H89712 also induced hl60 cells toward macrophagelike differentiation It was verified by flow cytometry that the percentage of positive CD14 expression in differentiated hl60 cells was about 9 times higher than that of the control at the dose of 108 mol/L and 20 times higher than that of the control at the dose of 105 mol/L respectively, and this action involved upregulation of MafB gene in hl60 leukemia cells On the other hand, H89712 inhibited proliferation of K562 leukemia cells and the inhibition of K562 leukemia cells in MTT assay was shown by 34% at the dose of 108 mol/L and 88% at the dose of 105 mol/L respectively The inhibition of K562 leukemia cells in colony assay was 53% at the dose of 108 mol/L and 100% at the dose of 105 mol/L respectively H89712 also induced K562 cells toward erythroidlike differentiation and it was verified by flow

关键词： 2-aminosteroid proliferation differentiation MafB Egr-1 hl-60 K562 leukemia

RECOMBINANT HUMAN G-CSF AND RETINOIC ACID IN SYNERGISTICALLY INDUCING GRANULOCYTE DIFFERENTIATION OF HUMAN PROMYELOCYTIC LEUKEMIC CELLS

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中华医学杂志(英文版) 1992年第9期105卷 707-712页

作者：黄非赵惠平高雪芝戴梅美范蕾蕾 Department of Hematology Affiliated Hospital Zhenjiang 212001 Department of Laboratory Medicine Affiliated Hospital Zhenjiang 212001 Department of Hematology Affiliated Hospital Nanjing Railway Medical College Nanjing 210009 Department of Hematology Affiliated Hospital Nanjing Railway Medical College Nanjing 210009Department of Hematology Affiliated Hospital Nanjing Railway Medical College Nanjing 210009

The effects of recombinant human G-CSF(rhG-CSF)and retinoic acid(RA)were studied on theproliferation and differentiation of hl-60 cells andhuman acute myeloid leukmic cells.Synergistic effecton granulocyte differentiation was observed whenhl-60 cells and primary acute promyelocyticleukemic cells were cocultured with RA plusrhG-CSF.rhG-CSF combined with RA increasedmore significantly the percentage of mature cells thanRA alone and greatly increased NBT reductionactivity(P<0.001).These results suggested thatproliferated effect of rhG-CSF on leukemic cells maybe important for inducing differentiation of myeloidleukemic cells.But this effect might expose the pa-tients to the risk of acute myeloblastic leukemia ifG-CSF was used alone.However,RA could not onlyrule out the latter situation but retain former merit aswell.The authors suggest that the combined use ofG-CSF with RA is probably a new approach to thetreatment of leukemia.

关键词： ACUTE MYELOID-LEUKEMIA INVITRO INDUCTION hl-60

The Effect of Curcumin on Mismatch Repair(MMR) Proteins hMSH2 and hMLH1 after Ultraviolet(UV) Irradiation on hl-60 Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2003年第2期23卷 124-126页

作者：陈燕吴裕丹陈文娟何静 The Institute of Hematology Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022

To understand the expression and effect of mismatch repair genes, hMSH2 and hMLH1, and to investigate anti-leukemic cell proliferation mechanism of curcumin, the levels of both genes were detected by multiple comparative RT-PCR. The protein of hMSH2 was determined by flow cy-tometry (FCM) and the gene mutation of hMSH2 and hMLH1 were detected by PCR-SSCP and mi-crosatellite instability assay. After UV irradiation, the gene expression of hMSH2 and hMLhl was not increased and showed no response. This phenomenon was not ascribed to gene mutation, because PCP-SSCP and microsatellite instability assay revealed no abnormal gel-shift band in both genes. After irradiation and addition of curcumin, the expression of hMSH2 mRNA increased and the cellular apoptotic rate also increased at the same time. The difference was statistically significant as compared with groups without addition of curcumin and control groups (P<0. 05). Our results suggested that when MMR system was inhibited by the same agents, curcumin can remove this suppression and switch to cellular apoptosis.

关键词： leukemia hl-60 mismatch repair gene curcumin

microRNA-181b targets MLK2 in hl-60 cells

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Science China(Life Sciences) 2010年第1期53卷 101-106页

作者： CHEN Hong1,CHEN Qun2,FANG Ming3 & MI Yan1 1 The First Affiliated Hospital of Harbin Medical University,Harbin 150101,China 2 Harbin Acheng District People’s Hospital,Harbin 150300,China 3 Graduate Department of Harbin Medical University,Harbin 150001,China The First Affiliated Hospital of Harbin Medical University Harbin China Harbin Acheng District People’s Hospital Harbin China Graduate department of Harbin Medical University Harbin China

microRNAs(miRNAs) play critical roles in many different cellular processes,including metabolism,apoptosis,differentiation and development.We showed miR-181b to be highly expressed in acute myeloid leukemia(AML).Furthermore,miR-181b contributed to proliferation of AML cells by targeting MLK2.Our results demonstrated that miR-181b plays an important role in the biology of AML and may be useful in developing therapies targeting miRNAs.

关键词： microRNA-181b MLK2 AML hl-60 proliferation apoptosis

Inhibition of NF-kappa B can enhance Fas-mediated apoptosis in leukemia cell line hl-60

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Frontiers of Medicine 2010年第3期4卷 323-328页

作者： Li WANG Shi ZHAO Hong-Xiang WANG Ping ZOU Department of Hematology The Central Hospital of WuhanWuhan 430014China Department of Hematology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China

This study explored the effects of nuclear factor-kappa B(NF-κB)inhibitor Bay 11-7082 on Fas/FasL system and Fas-mediated apoptosis in cell line hl-60 cells.The mRNA and protein levels of Fas,FasL,and X-linked inhibitor of apoptosis protein(XIAP)were detected by reverse transcription-polymerase chain reaction(RT-PCR)and flow cytometry(FCM);the level of sFasL was evaluated by enzyme-linked immunosorbent assay(ELISA);and apoptosis was determined by FCM.After treatment with Bay 11-7082,the mRNA and protein levels of FasL and XIAP in hl-60 cells were significantly lower than in the controls(P0.05).Apoptotic rate of hl-60 cells treated with Bay 11-7082 was significantly higher than in the controls(Phl-60 cells by downregulating FasL and XIAP levels.

关键词： nuclear factor-kappa B Fas/FasL system hl-60 Bay 11-7082

PROTEIN KINASE C IN THE PROMYELOCYTIC LEUKEMIA CELL DIFFERENTIATION OF GRANULOCYTES

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中华医学杂志(英文版) 1991年第1期104卷 32-35页

作者： Wu Xing-zhong Shao Guo-ying Chen Shu-rong Huang Meng-er Wang Zhen-yi Department of Hematology Jinling Hospital Nanjing 210002 Shanghai Institute of Hematology Shanghai Second Medical University Shanghai 200025

When the leukemia cells in 14 patients with acute promyelocytic leukemia （APL） were induced by all-trans retinoic acid （RA） treatment, the activity of protein kinase C （PKC） increased with the differentiation, in the control cells, the activity of PKC was only 47±39.3 pmol.mg/min, while it was 149.3±150.1 pmol.mg/min after differentiation. There were 90.1±7.2% promyelocytes in the control group, and 8.9±5.6% in the induction group. Therefore, we think that there is a close correlation between PKC and the differentiation of promyelocytic leukemia cells.

关键词： RETINOIC ACID INHIBITION INDUCTION hl-60