Bao C.,174th Hospital of Chinese Peoples Liberation Amiy |
Yaya Z.,174th Hospital of Chinese Peoples Liberation Amiy |
Cui D.L.,174th Hospital of Chinese Peoples Liberation Amiy |
Wei L.Y.,174th Hospital of Chinese Peoples Liberation Amiy |
Guo M.,174th Hospital of Chinese Peoples Liberation Amiy
Chinese Journal of Cancer Biotherapy | Year: 2014
Objective: To explore synergistic effects and the underlying mechanisms of liver kinase B1 (LKBl) or ser- ine-threonine kinase 11 (STK11) and metformin on proliferation and apoptosis of human cervical cancer cells using the HeLa cell line as a model. Methods: A recombinant plasmid LKB1-pEGFP-n1 was constructed. HeLa cells were transfected with this construct and the mock-vehicle pEGFP-n1 respectively. Transfectants were then treated with metfomiin. Cell viability was assessed by MTT assays, apoptosis and cycle progression by flow cytometry, and phosphorylation of AMRK, ACC and Rb (key players in the LKB1-AMPK signaling pathway) by Western blotting, 24 h after metfomiin treatment. Results: Both LKB1-pEGFP-n1 and the mock-vehicle pEGFP-n1 were successfully transfected into HeLa cells. After metfomiin treatment, IC50 was significantly lower in cells transfected with LKB1-pEGFP-n((2. 9 ± 0. 4) mmol/L than those transfected with pEGFP-n1 (7.8 ±1.3) mm ol/L and wild type HeLa cells (9.6±1.5)m mol/L (P <0.01), indicating a significant cell growth-inhibiting effect for LKB1. LKB1-pEGFP-n1 group showed a Gt phase arrest after treatment with metfomiin. In contrast, no cell cycle arrest was evident in wild type Hela cells or HeLa cells transfected with pEGFP-n1. After treatment with 15 mmol/L metfomiin, apoptosis rate was significantly higher in cells transfected with LKB1-pEGFP-n1 (28. 6 ± 2. 3)% than that in cells transfected withpEGFP-n1 (9.6 ± 1.6)% and wild type cells (17. 8 ± 1. 9)% (P < 0. 05). Phosphorylation AMPKα and ACC as increased but phosphorylation of Rb was decreased in cells transfected with LKB1-pEGFP-n1 as compared with untransfected cells and cells transfected with-pEGFP- n1. Conclusion: LH27 and metfomiin may affect the proliferation and apoptosis of cervical cancer cells in a coordinated manner, possibly involving the LKB1-AMPK signaling pathway.