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Expression of mTOR conduction pathway in human osteosarcoma MG-63 cells and their stem cells, and the inhibitory effect of different doses of rapamycin


Hao Wu
Xuelei Wang
Zhilin Cao
Mingdi Zheng
Zhongyuan Zhao
Yuchi Zhao
Jianzhong Zhang
Jianzhong Zhang
Gong Cheng

Abstract

Purpose: To investigate the expressions of rapamycin target protein (mTOR) conduction pathway in human osteosarcoma MG-63 cells and their stem cells, and to examine the inhibitory effect of different doses of rapamycin.
Methods: mTOR mRNA in osteosarcoma stem-like cells and human osteosarcoma MG-63 cells were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The cells were treated with different doses of rapamycin and divided into low dose group (0.5 mg), medium dose group (1.0 mg), high dose group (2.0 mg) and blank (control) group. Apoptosis and cell cycle of MG-63 cells were determined by flow cytometry, while proliferation of MG-63 cells up was assessed by CCK-8 kit.
Results: mTOR in human osteosarcoma MG-63 cells was significantly lower than that in osteosarcoma stem-like cells. Compared with the control group, mRNA expression levels of mTOR in MG-63 cells and osteosarcoma stem-like cells were significantly decreased after treatment with different concentrations of rapamycin (p < 0.05). MG-63 cells treated with various doses of rapamycin exhibited a significant decrease in their proliferation, compared with control group, while only the high rapamycin concentration group exhibited a significant decrease in osteosarcoma stem-like cell proliferation (p < 0.05). Treatment with rapamycin in MG-63 cells and osteosarcoma stem-like cells resulted in a significant increase in apoptosis, prolonged G0/G1 phase and shortened S phase (p < 0.05).
Conclusion: Rapamycin inhibits the expression of mTOR mRNA in osteosarcoma stem-like and MG-63 cells. It also inhibits the proliferation and cell cycle formation of osteosarcoma stem-like cells and MG-63 cells via mTOR signal pathway. These findings may provide a new target for the treatment of osteosarcoma.


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eISSN: 1596-9827
print ISSN: 1596-5996