Experimental Study
Osthole inhibits pancreatic cancer cell cycle progression and proliferation by inducing reactive oxygen species
Yao Jun, Wei Yijun, Zhou Fang, Yang Jian, Li Dechun, Zhou Jian
Published 2019-11-08
Cite as Chin J Exp Surg, 2019,36(11): 2035-2037. DOI: 10.3760/cma.j.issn.1001-9030.2019.11.034
Abstract
ObjectiveTo investigate the oxidative stress damage and mechanism of osthole (Ost) on panc-1 cells, and to provide experimental evidence for the treatment of pancreatic cancer with Chinese medicine.
MethodsPanc-1 cells were treated with different concentrations of Ost, and the proliferation of cells was detected by Alamar blue method. The cell viability was detected by trypan blue counting method. The cellular reactive oxygen species (ROS) content and cell cycle were detected by flow cytometry. The expression of Cyclin D1, Cyclin D3 and Notch1 in panc-1 cells was detected by Western blotting. The statistical method used one-way analysis of variance.
ResultsThe proliferation activity of panc-1 cells decreased under the action of Ost, and the concentration-dependent, drug inhibition rates of 10, 20, 40, 80, 120, 160, 200 μmol/L were (0.095±0.056)%, (0.164±0.033)% (0.201± 0.034)%, (0.267±0.037)%, (0.537±0.028)%, (0.732±0.015)%, (0.936±0.002)%. Flow cytometry results showed that ROS increased with increasing concentration and could be effectively reversed by antioxidant (NAC). The ROS value was 0 μmol/L: 17.700±0.795, 80 μmol/L: 19.750±0.101, NAC+ 80 μmol/L: 17.850±0.236. Trypan blue count showed a decrease in cell viability, 0 μmol/L: 100.000±7.536, 80 μmol/L: 65.610±4.351, NAC + 80 μmol/L: 76.110±1.603. After 48 h of Ost effect, G0/G1 phase increased, 0 μmol/L: (57.920±2.091)%, 120 μmol/L: (70.090±1.558)%; G2/M phase decreased, 0 μmol/L: (26.080±1.674)%, 120 μmol/L: (16.730± 0.781)%. This effect was also inhibited by NAC. The G0/G1 phase of the NAC+ 120 μmol/L group was (64.700±2.191)%, and the G2/M phase was (22.390±1.657)%. The relative expression levels of Cyclin D1 (40 μmol/L: 0.804±0.067, 80 μmol/L: 0.462±0.118) and Cyclin D3 (40 μmol/L: 0.845±0.035, 80 μmol/L: 0.372±0.096) were also shown to decrease at the protein level. In addition, Western blotting also showed that Ost can down-regulate the relative expression of Notch1 protein (40 μmol/L: 0.949±0.027; 80 μmol/L: 0.586±0.043). All results were statistically significant (P<0.05).
ConclusionOst can increase ROS in panc-1 cells in a concentration-dependent manner and inhibit proliferation by blocking cell cycle.
Key words:
Osthole; Pancreatic cancer; Oxidative stress; Proliferation
Contributor Information
Yao Jun
Department of General Surgery, the First Affiliated Hospital of Suzhou University, Suzhou 215006, China
Wei Yijun
Department of General Surgery, the First Affiliated Hospital of Suzhou University, Suzhou 215006, China
Zhou Fang
Department of General Surgery, the First Affiliated Hospital of Suzhou University, Suzhou 215006, China
Yang Jian
Department of General Surgery, the First Affiliated Hospital of Suzhou University, Suzhou 215006, China
Li Dechun
Department of General Surgery, the First Affiliated Hospital of Suzhou University, Suzhou 215006, China
Zhou Jian
Department of General Surgery, the First Affiliated Hospital of Suzhou University, Suzhou 215006, China
