Pseudolaric acid B induced cell cycle arrest, autophagy and senescence in murine fibrosarcoma L929 cell

Jing Hua Yu, Chun Yu Liu, Gui Bin Zheng, Li Ying Zhang, Ming Hui Yan, Wen Yan Zhang, Xian Ying Meng, Xiao Fang Yu

Research output: Contribution to journalArticlepeer-review

Abstract

Objective: PAB induced various cancer cell apoptosis, cell cycle arrest and senescence. But in cell line murine fibrosarcoma L929, PAB did not induce apoptosis, but autophagy, therefore it was thought by us as a good model to research the relationship of cell cycle arrest, autophagy and senescence bypass apoptosis. Methods: Inhibitory ratio was assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. Phase contrast microscopy visualized cell morphology. Hoechst 33258 staining for nuclear change, propidium iodode (PI) staining for cell cycle, monodansylcadaverine (MDC) staining for autophagy, and rodanmine 123 staining for mitochondrial membrane potential (MMP) were measured by fluorescence microscopy or flowcytometry. Apoptosis was determined by DNA ladder test. Protein kinase C (PKC) activity was detected by PKC assay kit. SA-β-galactosidase assay was used to detect senescence. Protein expression was examined by western blot. Results: PAB inhibited L929 cell growth in time-and dose-dependent manner. At 12 h, 80 μmol/L PAB induced obvious mitotic arrest; at 24 h, PAB began to induce autophagy; at 36 h, cell-treated with PAB slip into G1 cell cycle; and 3 d PAB induced senescence. In time sequence PAB induced firstly cell cycle arrest, then autophagy, then slippage into G1 phase, lastly senescence. Senescent cells had high level of autophagy, inhibiting autophagy led to apoptosis, and no senescence. PAB activated PKC activity to induce cell cycle arrest, autophagy and senescence, inhibiting PKC activity suppressed cell cycle arrest, autophagy and senescence. Conclusion: PAB induced cell cycle arrest, autophagy and senescence in murine fibrosarcoma L929 cell through PKC.

Original languageEnglish (US)
Pages (from-to)707-718
Number of pages12
JournalInternational journal of medical sciences
Volume10
Issue number6
DOIs
StatePublished - 2013

Keywords

  • Autophagy
  • Cell cycle arrest
  • Murine fibrosarcoma L929 cells
  • PKC
  • Pseudolaric acid B (PAB)
  • Senescence

ASJC Scopus subject areas

  • Medicine(all)

Fingerprint Dive into the research topics of 'Pseudolaric acid B induced cell cycle arrest, autophagy and senescence in murine fibrosarcoma L929 cell'. Together they form a unique fingerprint.

Cite this