title: [Subcellular re-distribution of F-actin in MCF7 cells upon inducible expression of CDKN2A/p16INK4a].
abstract: Objective To investigate the effect of CDKN2A/p16INK4a, a cyclin-dependent kinase inhibitor, on cell morphology and cytoskeleton in MCF7 breast cancer cells by tetracycline operon (Tet-on) inducible gene expression system. Methods MCF7 cells, genetically null for p16INK4aexpression, were sequentially infected with two viruses of Tet-on systems to make MCF7-pTet-on-p16INK4acells, which were induced to express p16NK4a by doxycycline (Dox) treatment. Cell growth was evaluated by cell counting. To observe the effect of p16INK4aon cell adhesion and cytoskeleton, intercellular adhesion and intracellular F-actin were examined by fluorescent staining with anti-E-cadherin antibody and phalloidin, respectively. Changes in cell morphology were analyzed by ImageJ software after microscopic imaging. Results Cell line expressing p16INK4aupon Dox induction was successfully made in MCF7 breast cancer cells. The inducible expression of p16INK4ainhibited cell proliferation while significantly increasing cell size as marked by prolonged contour lines. Furthermore, intracellular F-actin were enriched at the cortical peripheries of p16INK4a-expressing cells, which was different from those in control MCF7 cells where F-actin were enriched in cytoplasmic regions. Conclusion p16INK4aexpression in MCF7 breast cancer cells leads to increased cell size, weakened intercellular adhesion and cortical re-distribution of cytoskeletal F-actin.
authors.affiliation: School of Biological Science and Engineering, South China University of Technology, Guangzhou 510000; Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
authors.forename: Jianqing
authors.initials: J
authors.lastname: Liang
authors.affiliation: Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
authors.forename: Manna
authors.initials: M
authors.lastname: Wang
authors.affiliation: Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
authors.forename: Zhengrong
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
authors.forename: Long
authors.initials: L
authors.lastname: Yuan
authors.affiliation: Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
authors.forename: You
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: Department of Medical Oncology, Beijing Shijitan Hospital, Capital Medical University, Beijing Key Laboratory of Cancer Therapeutic Vaccine, Beijing 100038, China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Huang
authors.affiliation: Beijing Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China. *Corresponding authors, E-mail: sunq@bmi.ac.cn.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Sun
authors.affiliation: School of Biological Science and Engineering, South China University of Technology, Guangzhou 510000, China. *Corresponding authors, E-mail: 854415890@qq.com.
authors.forename: Jufang
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Biological Science and Engineering, South China University of Technology, Guangzhou 510000, China. *Corresponding authors, E-mail: xnwang88@163.com.
authors.forename: Xiaoning
authors.initials: X
authors.lastname: Wang
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism*; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle; D049109:Cell Proliferation; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics* / Q000378:metabolism; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism; D006801:Humans; D061986:MCF-7 Cells; D021381:Protein Transport
chemical_list: D000199:Actins; D015820:Cadherins; D019941:Cyclin-Dependent Kinase Inhibitor p16