Effects of cryptotanshinone on proliferation and apoptosis of human bladder carcinoma J82 cell line and its mechanisms / 中国基层医药

ABSTRACT

Objective:To study the effects of cryptotanshinone on proliferation and apoptosis of human bladder carcinoma J82 cells in vitro and its mechanisms.Methods:The J82 cells were treated with cryptotanshinone of different concentrations including 0, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0 and 8.0 μmol/L.After 48-hour treatment, the proliferation of J82 cells was determined by CCK-8 assay method.Flow cytometric analysis with Annexin V-FITC/PI staining was used to evaluate the apoptosis of J82 cells, and Western blotting was adopted to observe the protein expressions of p65, caspase-3, caspase-8 and caspase-9 related to proliferation and apoptosis.Results:CCK-8 results showed that the A values of 450nm-wavelength were as following: the control group (1.77±0.06), 0.5μmol/L group (1.78±0.08), 1.0μmol/L group (1.64±0.05), 2.0μmol/L group (1.48±0.12), 3.0μmol/L group (1.20±0.07), 4.0μmol/L group (0.93±0.10), 5.0μmol/L group (0.76±0.02), 8.0μmol/L group (0.05±0.01), and the A values were significantly different among the three groups ( F=329.83, P=0.00), there were statistically significant differences between any two groups except the 0.5 μmol/L group(all P<0.05). The early and total apoptosis rates were both significantly different among the three groups ( F=32.49, P=0.00; F=6.39; P=0.03), the early apoptosis rates of 3.0, 1.0μmol/L group were higher than that of the control group(all P<0.05), and the early apoptosis rate in the 3.0 μmol/L group was higher than that in the 1.0μmol/L group[(11.83±1.12)% vs.(7.01±1.84)%, t=3.73, P<0.05]. The expression of p65 protein decreased, while both the expressions of caspase-3 and caspase-9 proteins increased after treatment with cryptotanshinone. Conclusion:Cryptotanshinone can significantly inhibit proliferation and induce apoptosis of human bladder carcinoma J82 cells in vitro, probably via suppressing NF-κB signal pathway and activating mitochondrial pathway, respectively.