title: Tonglian Decoction () arrests the cell cycle in S-phase by targeting the nuclear factor-kappa B signal pathway in esophageal carcinoma Eca109 cells.
abstract: OBJECTIVE
  To investigate the anti-tumor activity and molecular mechanism of Tonglian Decoction (, TLD) on esophageal carcinoma Eca109 cells.
  
  
  METHODS
  Eca109 cells were treated with TLD and its separated formulae, including the clearing-heat and detoxification formula (Q), activating-blood and promoting-qi formula (H) and nourishing-yin and blood formula (Z). Cell proliferation was measured using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay, cell morphology was observed using a microscope, the cell cycle was measured using flow cytometry and the activity of the nuclear factor-kappa B (NF-κB) signal pathway was detected by Western blot.
  
  
  RESULTS
  The half maximal inhibitory concentrations of TLD, Q and H were 386, 771 and 729 mg/L, respectively. TLD, Q and H significantly inhibited cell proliferation, with 69.43%, 60.84% and 61.90% of treated cells in the G phase of the cell cycle. The percentage of cells in S phase increased significantly after treatment with TLD, Q, and H compared with the control group (P<0.05), and TLD showed the strongest effect. Z had no influence on the cell cycle compared with the control group (P>0.05). Western blot detection indicated slight differences in the inhibition of the NF-κB pathway by the different formulae. TLD formula strongly inhibited IKKβ, NF-κB, interleukin-6 and tumor necrosis factor-α expression compared with the control group.
  
  
  CONCLUSIONS
  TLD inhibited Eca109 cell proliferation by arresting cells in S phase. The possible mechanism might be related to inhibiting the NF-κB transduction cascade. The combination of the herbs found in the three separate formulae, H, Q and Z, work synergistically in TLD to produce the inhibitory effects of TLD treatment on Eca109 proliferation.
authors.affiliation: Basic Section, College of Chinese Medicine, North China University of Science and Technology, Tangshan, Hebei Province, 063000, China.
authors.forename: Yong-Sen
authors.initials: YS
authors.lastname: Jia
authors.affiliation: Institute of Orthopaedics and Traumatology, Zhejiang Chinese Medical University, Hangzhou, 310053, China. huxueqin@gmail.com.
authors.forename: Xue-Qin
authors.initials: XQ
authors.lastname: Hu
authors.affiliation: Basic Section, College of Chinese Medicine, North China University of Science and Technology, Tangshan, Hebei Province, 063000, China.
authors.forename: Ji-An
authors.initials: JA
authors.lastname: Li
authors.affiliation: Oncotherm Innovation and Trade Ltd., Pecs, 7322, Hungary.
authors.forename: Szasz
authors.initials: S
authors.lastname: Andras
authors.affiliation: Acupuncture Rehabilitation Foundation, Budapest, 1085, Hungary.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Hegyi
authors.affiliation: Traditional Chinese Medicine Hospital of Shenzhen, Shenzhen, Guangdong Province, 518020, China.
authors.forename: Bing-Sheng
authors.initials: BS
authors.lastname: Han
mesh_terms: D015153:Blotting, Western; D002452:Cell Count; D059447:Cell Cycle Checkpoints / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D004938:Esophageal Neoplasms / Q000378:metabolism* / Q000473:pathology*; D005434:Flow Cytometry; D006801:Humans; D020128:Inhibitory Concentration 50; D016328:NF-kappa B / Q000378:metabolism*; D016196:S Phase / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D004365:Drugs, Chinese Herbal; D016328:NF-kappa B; C000614301:tonglian decoction
keywords: Chinese medicine; Eca109 cells, Tonglian Decoction; cell cycle; esophageal carcinoma; nuclear factor-kappa B; proliferation