Jianqiang Wang1, Lijun Lao2, Hui Zhao1, Yuan Huang1. 1. Department of Gastroenterology, Second Hospital of Nanchang University Nanchang 330006, China. 2. Department of General Surgery, First Affiliated Hospital, Liaoning Medical University Jinzhou 121001, China.
Abstract
OBJECTIVE: This study aimed to investigate the effects of serine threonine kinase Pim-3 on the growth of HepG2 cells and to explore the role of STAT3 signaling pathway. METHODS: Synthetic Pim-3shRNA and negative control shRNA were independently transfected into HepG2 cells in the presence of Lipofectamine(TM) 2000. Cells were divided into 4 groups: Pim-3 shRNA group, negative control group, liposome control group, and blank control group. Flow cytometry was performed to detect the apoptosis of these cells; RT-PCR was employed to detect the mRNA expression of Pim-3; Western blot assay was done to measure the protein expression of Pim-3, STAT3, pSTAT3(Tyr705), Bcl-Xl, Bad and pBad(Ser112). RESULTS: When compared with blank control group, liposome group and negative control group, the apoptosis index increased and the protein expression of Pim-3, pSTAT3(Tyr705), Bcl-Xl and pBad(Ser112) and the Pim-3 mRNA expression reduced in the Pim-3 shRNA group, but the protein expression of STAT3 and Bad was comparable among groups. CONCLUSION: Pim-3 shRNA may down-regulate pSTAT3(Tyr705) and pBad(Ser112) protein expression to inhibit the proliferation of liver cancer cells and Pim-3 may serve as a target for the treatment of liver cancer.
OBJECTIVE: This study aimed to investigate the effects of serine threonine kinase Pim-3 on the growth of HepG2 cells and to explore the role of STAT3 signaling pathway. METHODS: Synthetic Pim-3shRNA and negative control shRNA were independently transfected into HepG2 cells in the presence of Lipofectamine(TM) 2000. Cells were divided into 4 groups: Pim-3 shRNA group, negative control group, liposome control group, and blank control group. Flow cytometry was performed to detect the apoptosis of these cells; RT-PCR was employed to detect the mRNA expression of Pim-3; Western blot assay was done to measure the protein expression of Pim-3, STAT3, pSTAT3(Tyr705), Bcl-Xl, Bad and pBad(Ser112). RESULTS: When compared with blank control group, liposome group and negative control group, the apoptosis index increased and the protein expression of Pim-3, pSTAT3(Tyr705), Bcl-Xl and pBad(Ser112) and the Pim-3 mRNA expression reduced in the Pim-3 shRNA group, but the protein expression of STAT3 and Bad was comparable among groups. CONCLUSION:Pim-3 shRNA may down-regulate pSTAT3(Tyr705) and pBad(Ser112) protein expression to inhibit the proliferation of liver cancer cells and Pim-3 may serve as a target for the treatment of liver cancer.
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