Moleecular Mechanism of K562 Senescence in Chronic Myeloigenous Leukemia Cells Induced by Targeting of P16INK4a Protein by Bone Marrow Mesenchymal Stem Cells

Abstract

Abstract:

〔Abstract〕 Objective： To explore the mechanism of mouse bone marrow mesenchymal stem cell（BMMSC） inducing K562 cells aging through p16INK4a protein. Methods：Mesenchymal stem cells were isolated and purified from mouse bone marrow. The changes of the K562 cell cycle were detected and the surface markers were identified by flow cytometry. The ability of adipogenic and osteogenic differentiation was identified by oil red O and alizarin red staining. The proliferation of K562 cells was detected by CCK-8 method. The senescence of K562 cells was detected by SA-β-Gal staining. The expression of p16INK4a protein was detected by Western blot. Results： BMMSC showed fibroblast-like adherent growth， with low expression of hematopoietic stem cell marker CD45 and high expression of stem cell surface markers CD29 and CD44 ， which had osteogenic and adipogenic differentiation ability. BMMSC could inhibit the proliferation of K562 cells， block K562 cell cycle in G0/G1 phase， and improve SA-β-Gal activity and p16INK4a protein expression. Conclusion： BMMSC can induce the senescence of K562 cells by up-regulating the level of p16INK4a protein.

Key words:

"> font-size:10.5pt, ">bone marrow mesenchymal stem cell, senescence, leukemia, K562 cells, p16INK4a protein

CLC Number:

Zhou Wen, Tang Ziyun, He Siyue, Liu Xiaohu, Zhou Yue.

Moleecular Mechanism of K562 Senescence in Chronic Myeloigenous Leukemia Cells Induced by Targeting of P16INK4a Protein by Bone Marrow Mesenchymal Stem Cells [J]. Journal of Dali University, 2022, 7(10): 39-46.

References

〔1〕 ROWLEY J D. LETTER： A New Consistent Chromosomal Abnormality in Chronic Myelogenous Leukaemia Identified by Quinacrine Fluorescence and Giemsa Staianing〔J〕. Nature， 1973，243（5405）：290-293.

〔2〕 SASAKI K， STROM S S， O'BRIEN S， et al. Relative Survival in Patients with Chronic-Phase Chronic Myeloid Leukaemia in the Tyrosine-Kinase Inhibitor Era： Analysis of Patient Data from Six Prospective Clinical Trials〔J〕. Lancet Haematol， 2015，2（5）：e186-e193.

〔3〕 ROEKER L E， DREGER P， BROWN J R， et al. Allogeneic Stem Cell Transplantation for Chronic Lymphocytic Leukemia in the Era of Novel Agents〔J〕. Blood Adv， 2020，4（16）：3977-3989.

〔4〕 DUARTE D， HAWKINS E D， LO CELSO C. The Interplay of Leukemia Cells and the Bone Marrow Microenvironment〔J〕. Blood， 2018，131（14）：1507-1511.

〔5〕 FATHI E， FARAHZADI R， VALIPOUR B， et al. Cytokines Secreted from Bone Marrow Derived Mesenchymal Stem Cells Promote Apoptosis and Change Cell Cycle Distribution of K562 Cell Line as Clinical Agent in Cell Transplantation〔J〕. PLoS One，2019，14（4）：e0215678.

〔6〕 FONSEKA M， RAMASAMY R， TAN B C， et al. Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells （hUCB-MSC） Inhibit the Proliferation of K562 （Human Erythromyeloblastoid Leukaemic Cell Line）〔J〕. Cell Biol Int， 2012， 36（9）： 793-801.

〔7〕 FATHI E， VALIPOUR B， SANAAT Z， et al. Interleukin-6，-8， and TGF-β Secreted from Mesenchymal Stem Cells Show Functional Role in Reduction of Telomerase Activity of Leukemia Cell via Wnt5a/β-Catenin and P53 Pathways〔J〕. Adv Pharm Bull， 2020， 10（2）： 307-314.

〔8〕 LIU J Y， SOUROULLAS G P， DIEKMAN B O， et al. Cells Exhibiting Strong P16INK4a Promoter Activation in vivo Display Features of Senescence〔J〕. Proc Natl Acad Sci USA， 2019， 116（7）： 2603-2611.

〔9〕 LIU Y， SANOFF H K， CHO H， et al. Expression of P16（INK4a） in Peripheral Blood T-Cells Is a Biomarker of Human Aging〔J〕. Aging Cell， 2009， 8（4）： 439-448.

〔10〕 YAO W J， LIANG Y， CHEN K， et al. Changes of Biophysical Behavior of K562 Cells for P16 Gene Transfer〔J〕. Clin Hemorheol Microcirc， 2002， 27（3/4）： 177-183.

〔11〕 RUI H B， SU J Z. Co-Transfection of P16（INK4a） and P53 Genes into the K562 Cell Line Inhibits Cell Proliferation〔J〕. Haematologica， 2002， 87（2）： 136-142.

〔12〕 KREVVATA M， SILVA B C， MANAVALAN J S， et al. Inhibition of Leukemia Cell Engraftment and Disease Progression in Mice by Osteoblasts〔J〕. Blood， 2014， 124（18）： 2834-2846.

〔13〕 XIA C X， WANG T J， CHENG H， et al. Mesenchymal Stem Cells Suppress Leukemia via Macrophage-Mediated Functional Restoration of Bone Marrow Microenvironment〔J〕. Leukemia， 2020， 34（9）： 2375-2383.

〔14〕 POTTEN C S， LOEFFLER M. Stem Cells： Attributes， Cycles， Spirals， Pitfalls and Uncertainties. Lessons for and from the Crypt〔J〕. Development，1990，110（4）：1001-

1020.

〔15〕 PAL B， DAS B. In vitro Culture of Naïve Human Bone Marrow Mesenchymal Stem Cells： A Stemness Based Approach〔J〕. Front Cell Dev Biol， 2017， 5：69.

〔16〕 DIMRI G P， LEE X， BASILE G， et al. A Biomarker that Identifies Senescent Human Cells in Culture and in Aging Skin in vivo〔J〕. Proc Natl Acad Sci USA，1995，92（20）： 9363-9367.

〔17〕 HE S H， SHARPLESS N E. Senescence in Health and Disease〔J〕. Cell， 2017， 169（6）： 1000-1011.

〔18〕 RAYESS H， WANG M B， SRIVATSAN E S. Cellular Senescence and Tumor Suppressor Gene P16〔J〕. Int J Cancer， 2012， 130（8）： 1715-1725.

〔19〕 MASSAGUÉ J. G1 Cell-Cycle Control and Cancer〔J〕. Nature， 2004， 432（7015）： 298-306.

〔20〕 BAKER D J， WIJSHAKE T， TCHKONIA T， et al. Clearance of P16Ink4a-Positive Senescent Cells Delays Ageing-Associated Disorders〔J〕.Nature，2011，479（7372）： 232-236.

〔21〕 LAPAK K M， BURD C E. The Molecular Balancing Act of P16（INK4a） in Cancer and Aging〔J〕. Mol Cancer Res， 2014， 12（2）： 167-183.

〔22〕 MINAMI R， MUTA K， UMEMURA T， et al. P16（INK4a） Induces Differentiation and Apoptosis in Erythroid Lineage Cells〔J〕. Exp Hematol， 2003，31（5）： 355-362.

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