Apoptosis of human prostate cancer cells induced by marine actinomycin X2 through the mTOR pathway compounded by MiRNA144
research target：miRNA144
Periodicals：Anti-cancer drugs
IF：1.784
Cooperative Unit：Guangzhou Military Region General Hospital, South China University of Technology
Time of publication：September，2015
Summary
The present study aimed to determine whether actinomycin X2 (AX2) intercepted the mTOR/PTEN/PI3K/Akt signaling pathway to inhibit human prostate cancer cells (PC-3) in vitro. The effects of AX2 on mTOR, PTEN, PI3K, and Akt at the protein level and mRNA were determined by western blotting and real-time reverse transcription-PCR (RT-PCR), respectively. Concurrently, the effects of AX2 on expression levels of MiRNA144 and MiRNA126 in PC-3 were measured by real-time RT-PCR. The association of MiRNA144 with 3′- UTR of mTOR was identified using the Dual-Luciferase Reporter Gene System. The direct effect of MIRNA144 on the mTOR/PTEN/PI3K/Akt pathway was determined by real-time RT-PCR and western blotting. Apoptosis of PC-3 cells induced by AX2 was determined by MTT and flow cytometry. The results indicated that mTOR/PTEN/PI3K/ Akt were decreased and PTEN was increased by AX (1, 10 µmol/l) at protein and mRNA levels in a dose-dependent manner. MiRNA144 was decreased, whereas MiRNA126 was increased by AX2. MiRNA144 associated with 3′-UTR of mTOR was corroborated. Overexpression of MiRNA144 decreased mTOR, but did not affect PTEN, PI3K, or Akt. The proliferation rates of AX2 on PC-3 cells were decreased. This suggests that AX2 induces apoptosis of PC-3 cells by meddling in the mTOR/PTEN/PI3K/Akt signaling pathway, but compounded by MiRNA144, and AX2 and MiRNA144 AQ2 intercepts it in different ways but crosses on mTOR.
Partial results of cooperation
1. The gene expression levels of mTOR-PTEN-PI3K-Akt signaling pathway were affected differently by AX2. By qPCR and WB detection, it was found that when PC-3 cells were treated with 10 umol/l AX2, the expression levels of mTOR, PI3K and Akt were significantly decreased, on the contrary, the expression level of PTEN was significantly increased.
2. The expression of miRNA126 and miRNA144 in PC-3 cells treated with different concentrations of AX2.
3. Dual-luciferase reporter gene to detect the interaction between mTOR gene and miRNA144.
4. When PC-3 cells were treated with miRNA144 mimic, the expression of mTOR mRNA was significantly decreased, but there was no significant change in PTEN, PI3K and AKT.
5. AX2 treatment was detected by flow cytometry as a result of apoptosis.
References
Liu J, Xie S, Wu Y, et al. Apoptosis of human prostate cancer cells induced by marine actinomycin X2 through the mTOR pathway compounded by MiRNA144.[J]. Anti-cancer drugs, 2016, 27(3).
