Tianjin Medical Journal

Tianjin Medical Journal ›› 2020, Vol. 48 ›› Issue (9): 801-806.doi: 10.11958/20200238

• Cell and Molecular Biology •     Next Articles

Effects of different concentrations of minocycline on the proliferation, autophagy and apoptosis of human glioma cells and its mechanism

HAN Li-min1, YAN Wan-yue2, LI Qiao-qiao2, LI Ke2, LIU Li2, ZHAO Hai-long1△ #br#   

  1. 1 Department of Pathophysiology, School of Basic Medical Sciences, Zunyi Medical University, Zunyi 563000, China;
    2 the First Clinical Institute of Zunyi Medical University
  • Received:2020-01-21 Revised:2020-05-11 Published:2020-09-15 Online:2020-09-15

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Abstract:

Abstract：Objective To investigate the effects and mechanism of minocycline at different concentrations on the
proliferation and apoptosis of human glioma U87 and LN229 cells. Methods (1) U87 and LN229 cells were divided into
three groups: control group (DMSO treatment), 5 μmol/L minocycline group and 10 μmol/L minocycline group. After 72
hours of treatment, the level of cell proliferation was detected by MTT method, the expression of autophagy LC3B was
detected by immunofluorescence labeling and the expressions of autophagy and apoptosis-related proteins were detected by
Western blot assay. (2) SIRT1-shRNA vector was constructed to observe the effects of different concentrations of
minocycline on the autophagy of U87 and LN229 cells after knockdown of SIRT1 expression. Results (1) Compared with
the control group, the MTT results showed that the proliferation of glioma cells was significantly inhibited in 5 μmol/L and 10
μmol/L minocycline groups, and immunofluorescence staining showed that the development of autophagy labeled protein
LC3B was increased. Western blot results showed that the level of mTOR was significantly decreased, and the levels of Atg5,
p-AMPK α and SIRT1 were significantly increased (P＜0.05). Western blot assay showed that the levels of Bcl-2 and pp70s6k were significantly decreased and the activated Caspase-3 level was significantly increased in 10 μmol/L minocycline group compared with those of control group (P＜0.05), but there were no significant changes in the levels of Bcl-2 and pp70s6k proteins in 5 μmol/L minocycline group (P＞0.05). (2) After knocking down the expression of SIRT1, the expression
levels of mTOR and LC3B were significantly lower in shSIRT1 group than those of control group, while after minocycline
treatment, the expression of mTOR increased significantly, while the expression level of LC3B only partially recovered.
Conclusion Minocycline of 5 and 10 μmol/L can effectively inhibit the growth of human glioma U87 and LN229 cells,
which may be related to the AMPK/SIRT1 pathway induced cell autophagy and p70s6k/Bcl-2 pathway induced apoptosis.

Key words: minocycline, glioma, cell proliferation, apoptosis, autophagy, AMP-activated protein kinases, SIRT1

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