Effect of PI3K-mediated autophagy in human osteosarcoma MG63 cells on sensitivity to chemotherapy with cisplatin

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• 作者：Xu-Dong Miao¹ ; Le Cao¹ ; Qiang Zhang¹ ; Xiao-Ying Hu² ; Yi Zhang² ; ^{zhangyi00934@163.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Autophagy ; Osteosarcoma ; LC3 ; Cisplatin ; Proliferation
• 刊名：Asian Pacific Journal of Tropical Medicine
• 出版年：2015
• 出版时间：September 2015
• 年：2015
• 卷：8
• 期：9
• 页码：731-738
• 全文大小：1625 K

文摘

To investigate the influence of autophagy on sensitivity to the chemotherapy drug cisplatin (DDP) and the role of PI3K in autophagy.

Methods

MTT methods and flow cytometer, with rapamycin up-regulating the autophagy and 3-MA down-regulating the autophagy, were employed to measure the proliferation inhibition rate on DDP-treated osteosarcoma cells and the change in cell cycle. The expression of intracellular protein was detected by Western blot. The autophagy of MG63 cell was observed using fluorescence microscope and transmission electron microscope.

Results

Western blot showed that basic autophagy level of MG63 cell was significantly lower than that of hFOB cell. MTT test revealed that the cell proliferation inhibition rate in the group treated with rapamycin and DDP, group treated with 3-MA and DDP, and group only treated with DDP was significantly different. It was demonstrated by the flow cytometry that in group treated with DDP, inhibition on autophagy can increase the cell numbers in G₁ phase and reduce the cell numbers in S phase of cell cycle. Increase of autophagosome in MG63 cytoplasm was observed under fluorescence microscope.

Conclusions

Up-regulating the autophagy significantly reduced the sensitivity of MG63 cell to chemotherapy with DDP. DDP induced autophagy of MG63 cell and blocked the cell cycle at G₁ phase.