首页|LAIR-1通过阻断JAK2 V617F突变的人HEL细胞JAK/STAT和PI3K/AKT/mTOR信号通路抑制其增殖并促进其凋亡

LAIR-1通过阻断JAK2 V617F突变的人HEL细胞JAK/STAT和PI3K/AKT/mTOR信号通路抑制其增殖并促进其凋亡

扫码查看
原文链接
  • 万方数据
  • 维普
目的 研究人白细胞相关免疫球蛋白样受体1(LAIR-1)对Janus激酶2(JAK2)V617F突变的人急性髓系白血病HEL细胞JAK/信号转导子与转录激活子(STAT)和磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/AKT/mTOR)信号通路的调节作用，以及对细胞增殖和凋亡的影响。方法 采用反转录PCR和基因测序鉴定JAK2 V617F突变;应用免疫共沉淀和Western blot法鉴定LAIR-1募集的蛋白酪氨酸磷酸酶(PTP)种类;采用CCK-8法检测HEL细胞的增殖;采用异硫氰酸荧光素标记的膜联素V/碘化丙啶(annexin V-FITC/PI)双标记结合流式细胞术检测HEL细胞的凋亡率;采用Western blot法检测JAK/STAT和PI3K/AKT/mTOR通路蛋白酪氨酸磷酸化水平及细胞周期蛋白D1(cyclin D1)、Bcl2相关X蛋白(BAX)和B细胞淋巴瘤因子2(Bcl2)的蛋白表达。结果 在JAK2 V617F突变的HEL细胞中，LAIR-1与其配体胶原蛋白结合后可募集含Src同源域2磷酸酶2(SHP-2);LAIR-1可以下调HEL细胞JAK2、STAT1、STAT3、STAT5、AKT和mTOR的蛋白酪氨酸磷酸化水平，并能够显著抑制cyclin D1和Bcl2的表达，而对BAX的表达水平未见显著影响;LAIR-1能够明显抑制HEL细胞的增殖，促进HEL细胞凋亡。结论 在JAK2 V617F突变的人白血病HEL细胞中，LAIR-1可通过募集SHP-2抑制JAK/STAT和PI3K/AKT/mTOR信号通路的活化，进而抑制HEL细胞的增殖，促进细胞凋亡。
leukocyte-associated immunoglobulin-like receptor 1(LAIR-1)inhibits proliferation and promotes apoptosis of human HEL cells with JAK2 V617F mutation by blocking the JAK/STAT and PI3K/AKT signaling pathways
Objective To investigate the role of human leukocyte-associated immunoglobulin-like receptor-1(LAIR-1)in the regulation of Janus kinase/signal transducers and activators of transcription(JAK/STAT)and phosphatidylinositol 3 kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)signaling pathways in human acute myeloid leukemia HEL cells carrying the JAK2 V617F mutation,along with its effects on cell proliferation and apoptosis.Methods The JAK2 V617F mutation was identified using reverse transcription PCR and gene sequencing.The protein phosphatase(PTP)recruited by LAIR-1 was determined through co-immunoprecipitation and Western blot analysis.The proliferation of HEL cells was detected by CCK-8 assay.The apoptosis rate of HEL cells was detected by flow cytometry with annexin V-FITC/PI labeling.Western blot analysis was employed to assess the phosphorylation status of proteins involved in the JAK/STAT and PI3K/AKT/mTOR pathways,as well as the expression levels of cyclinD1,B cell lymphoma 2(Bcl2),and Bcl2 associated X protein(BAX).Results In HEL cells containing the JAK2 V617F mutation,LAIR-1 was observed to recruit SH2-containing protein tyrosine phosphatase 2(SHP-2)upon binding with its ligand collagen.Moreover,LAIR-1 downregulated the tyrosine phosphorylation levels of JAK2,STAT1,STAT3,STAT5,AKT and mTOR and significantly reduced the expression of cyclin D1 and Bcl2,while having no effect on the expression of BAX.In addition,LAIR-1 exhibited a significantly inhibitory effect on cell proliferation and promoted apoptosis in HEL cells.Conclusion In HEL cells with JAK2 V617F mutation,LAIR-1 can inhibit the activation of JAK/STAT and PI3K/AKT/mTOR signaling pathways by recruiting SHP-2,thereby inhibiting the proliferation of HEL cells and promoting cell apoptosis.

myeloproliferative neoplasia(MPN)leukocyte-associated immunoglobulin-like receptor-1(LAIR-1)JAK2 V617F mutationJanus kinase(JAK)signal transducers and activators of transcription(STAT)phosphatidylinositol 3 kinase(PI3K)protein kinase B(AKT)

樊翠、张娅薇、杨蕊、吴肖婕、周嘉迪、薛江楠

展开 >

滨州医学院基础医学院免疫学教研室,山东烟台 264000

临沂市中医医院超声医学科,山东临沂 276000

滨州市人民医院中心实验室,山东滨州 256610

陕西中医药大学附属医院检验科,陕西咸阳 712000

展开 >

骨髓增殖性肿瘤 白细胞相关免疫球蛋白样受体1(LAIR-1) JAK2 V617F突变 Janus激酶(JAK) 信号转导子与转录激活子(STAT) 磷脂酰肌醇3激酶(PI3K) 蛋白激酶B(AKT)

国家自然科学基金

81273200

2024

10.13423/j.cnki.cjcmi.009736

细胞与分子免疫学杂志
中国免疫学会,第四军医大学

细胞与分子免疫学杂志

CSTPCD北大核心
影响因子：0.817
ISSN：1007-8738
年,卷(期)：2024.40(3)
  • 25