title: Paeoniflorin inhibits human gastric carcinoma cell proliferation through up-regulation of microRNA-124 and suppression of PI3K/Akt and STAT3 signaling.
abstract: OBJECTIVE
  To examine the potential anti-tumor activity of paeoniflorin in the human gastric carcinoma cell line MGC-803.
  
  
  METHODS
  Cell viability and cytotoxic effects in MGC-803 cells were analyzed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and lactate dehydrogenase assay, respectively. Cell apoptosis of MGC-803 cells was measured using flow cytometry, DAPI staining assay and caspase-3 activity assay. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression of microRNA-124 (miR-124) in response to paeoniflorin. The expression of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), phospho-Akt (p-Akt) and phospho-signal transducer and activator of transcription 3 (p-STAT3) were also measured by quantitative RT-PCR and Western blot analysis in normal, miR-124 and anti-miR-124 over-expressing MGC-803 cells, treated with paeoniflorin.
  
  
  RESULTS
  Paeoniflorin was found to inhibit MGC-803 cell viability in a dose-dependent manner. Paeoniflorin treatment was associated with the induction of apoptosis and caspase-3 activity in MGC-803 cells. Paeoniflorin treatment significantly increased miR-124 levels and inhibited the expression of PI3K, Akt, p-Akt and p-STAT3 in MGC-803 cells. Interestingly, the over-expression of miR-124 inhibits PI3K/Akt and phospho-STAT3 expressions in MGC-803 cells. PI3K agonist (IGF-1, 1 μg/10 μL) or over-expression of STAT3 reversed the effect of paeoniflorin on the proliferation of MGC-803 cells. Over-expression of anti-miR-124 in MGC-803 cells reversed paeoniflorin-induced up-regulation.
  
  
  CONCLUSIONS
  In summary, the in vitro data suggest that paeoniflorin is a potential novel therapeutic agent against gastric carcinoma, which inhibits cell viability and induces apoptosis through the up-regulation of miR-124 and suppression of PI3K/Akt and STAT3 signaling.
authors.affiliation: Yong-Bin Zheng, Gao-Chun Xiao, Shi-Lun Tong, Yu Ding, Qiu-Shuang Wang, Sheng-Bo Li, Zhi-Nan Hao, Department of Gastrointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.
authors.forename: Yong-Bin
authors.initials: YB
authors.lastname: Zheng
authors.affiliation: Yong-Bin Zheng, Gao-Chun Xiao, Shi-Lun Tong, Yu Ding, Qiu-Shuang Wang, Sheng-Bo Li, Zhi-Nan Hao, Department of Gastrointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.
authors.forename: Gao-Chun
authors.initials: GC
authors.lastname: Xiao
authors.affiliation: Yong-Bin Zheng, Gao-Chun Xiao, Shi-Lun Tong, Yu Ding, Qiu-Shuang Wang, Sheng-Bo Li, Zhi-Nan Hao, Department of Gastrointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.
authors.forename: Shi-Lun
authors.initials: SL
authors.lastname: Tong
authors.affiliation: Yong-Bin Zheng, Gao-Chun Xiao, Shi-Lun Tong, Yu Ding, Qiu-Shuang Wang, Sheng-Bo Li, Zhi-Nan Hao, Department of Gastrointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Yong-Bin Zheng, Gao-Chun Xiao, Shi-Lun Tong, Yu Ding, Qiu-Shuang Wang, Sheng-Bo Li, Zhi-Nan Hao, Department of Gastrointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.
authors.forename: Qiu-Shuang
authors.initials: QS
authors.lastname: Wang
authors.affiliation: Yong-Bin Zheng, Gao-Chun Xiao, Shi-Lun Tong, Yu Ding, Qiu-Shuang Wang, Sheng-Bo Li, Zhi-Nan Hao, Department of Gastrointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.
authors.forename: Sheng-Bo
authors.initials: SB
authors.lastname: Li
authors.affiliation: Yong-Bin Zheng, Gao-Chun Xiao, Shi-Lun Tong, Yu Ding, Qiu-Shuang Wang, Sheng-Bo Li, Zhi-Nan Hao, Department of Gastrointestinal Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.
authors.forename: Zhi-Nan
authors.initials: ZN
authors.lastname: Hao
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002277:Carcinoma / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D015972:Gene Expression Regulation, Neoplastic; D005960:Glucosides / Q000494:pharmacology*; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D039821:Monoterpenes / Q000494:pharmacology*; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism*; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D050796:STAT3 Transcription Factor / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D013274:Stomach Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D013997:Time Factors; D014162:Transfection; D015854:Up-Regulation; D006027:Glycosides
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D053148:Caspase 3; D005960:Glucosides; D035683:MicroRNAs; D039821:Monoterpenes; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt; D050796:STAT3 Transcription Factor; C509207:MIRN124 microRNA, human; C494086:STAT3 protein, human; C015423:peoniflorin; D006027:Glycosides; C505407:CASP3 protein, human
keywords: Akt; Gastric cancer; MicroRNA-124; Paeoniflorin; Phosphatidylinositol 3-kinase; Signal transducer and activator of transcription 3