TIAN Long-fu, ZHANG Qi, WANG Bo-tao, et al. Mechanism of Sanguinarine in Inducing Apoptosis in Murine Pancreatic Cancer Cells by Inhibiting PI3K/Akt Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(19): 166-171.
DOI:
TIAN Long-fu, ZHANG Qi, WANG Bo-tao, et al. Mechanism of Sanguinarine in Inducing Apoptosis in Murine Pancreatic Cancer Cells by Inhibiting PI3K/Akt Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(19): 166-171. DOI: 10.13422/j.cnki.syfjx.20181719.
Mechanism of Sanguinarine in Inducing Apoptosis in Murine Pancreatic Cancer Cells by Inhibiting PI3K/Akt Signaling Pathway
Objective: To investigate the effect of sanguinarine on pancreatic cancer cell apoptosis and its signal pathway regulation mechanism. Method: After Panc02 cells were treated with different concentrations of sanguinarine(2
4
6 μmol·L-1) for different periods of time(24
48
72 h)
the inhibition of cell growth was determined by 3-(4
5-dimethyl-2-thiazolyl)-2
5-diphenyl-2H-tetrazolium bromide (MTT) assay. The effect of sanguinarine on cell apoptosis was determined by phospholipid binding protein V/propidium iodide(Annexin V-FITC/PI) double staining flow cytometer assay. The changes in mitochondrial membrane potential were examined by cyanine dye (JC-1) staining assay. The expressions of protein B-lymphoblast-2 (Bcl-2)
Bcl-2 related X protein(Bax)
total protein kinase B (Akt)
phosphorylated protein kinase B (p-Akt)
total phosphatidylinositol 3 kinase (PI3K)
phosphorylated phosphatidylinositol 3-kinase(p-PI3K) were measured by Western blot analysis. Result: Compared with the blank group
the growth inhibition rate of Panc02 cells increased gradually with the rise of the drug concentration
after being treated with different concentrations of sanguinarine for the same period of time (P<0.05). Compared with the blank group
the inhibition rate of cell growth increased gradually with the rise of time
after being treated with the same concentration for different periods of time (P<0.05). Compared with the blank group
the apoptosis rate was not obviously increased at the dose of the 2 μmol·L-1 sanguinarine. Compared with the blank group
the apoptotic rate was increased at the dose of the 4
6 μmol·L-1 sanguinarine (P<0.05). Compared with the blank group
there was no change in the red and green fluorescence at the dose of 2 μmol·L-1
but a decrease in red fluorescence
an increase in green fluorescence and a reduction in Panc02 mitochondrial membrane potential at the dose of 4
6 μmol·L-1 sanguinarine. Compared with the blank group
the expressions of Bax
Bcl-2
p-PI3K
p-Akt
Akt and PI3K were not changed at the dose of the 2 μmol·L-1 sanguinarine. The expression of Bax was increased
while the expressions of Bcl-2 and PI3K/Akt signaling pathway in key proteins p-PI3K
p-Akt were decreased at the dose of 6 μmol·L-1 sanguinarine (P<0.05). Conclusion: Sanguinarine can effectively induce the apoptosis of pancreatic cancer Panc02 cells through the mitochondrial apoptotic pathway by inhibiting the PI3K/Akt signaling pathway.
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