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Effect of curcumin on extrinsic apoptosis pathway in human multiple myeloma cell ARH-77
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Abstract:
Objective:To investigate the influence of curcumin (Cur) on the extrinsic apoptosis pathway of human multiple myeloma cell line ARH-77. Methods: ARH-77 cells were treated with Cur at 6.25, 12.5, 25, 50, 100 and 200 μmol/L. At 12, 24 and 48 h after treatment, cell viability was analyzed by MTT assay and growth inhibition was accordingly calculated. At 24 h after treatment, changes in the cell morphology were assessed by Hoechst 33258 staining, cell cycle progression and levels of Fas/Fasl and TRAIL/TRAIL-R were analyzed by flow cytometry, and the activity of caspase 8 was determined by colorimetry. Results: Cur significantly inhibited the growth of ARH-77 cells in a time- and dose-dependent manner. At 24 h after treatment, Cur induced apoptosis in ARH-77 cells in a dose-dependent manner; the percentage of apoptotic cells was (10.35±0.35)% at 6.25 μmol/L, (1435±1.34)% at 12.5 μmol/L and (36.65±106)% at 25 μmol/L, significantly higher than that in untreated control cells (\[3.83±0.32\]%, P<0.01). Apoptotic bodies and cell cycle arrest at the G0/G1 phase were seen in ARH-77cells treated with 25 μmol/L Cur. Caspase 8 activity was significantly higher in ARH-77 cells treated with Cur at 6.26 μmol/L(0.223±0.018), 12.5 μmol/L (0.263±0.019), or 25.0 μmol/L (0.240±0.035) than in untreated control cells (0.154±0.007) (P<0.05). Compared with the non-treatment control, 24 h Cur treatment at 6.25 μmol/L significantly increased the protein levels of Fas (\[99.05±0.49\]% vs \[92.10±0.7\]%, P＝0.000), FasL (\[9.05±0.78\]% vs \[1.73±1.19\]%, P＝0008), TRAIL (\[1.35±0.07\]% vs \[0.55±0.07\]%, P＝0.008), DR4, DcR1 and DcR2 but significantly decreased DR5 (\[0.95±0.07\]% vs \[7.70±0.29]%, P＝0.001). The effect of Cur on DcR1 (\[4.35±1.20\]% vs \[14.25±0.21\]%, P＝0.008) and DcR2 (\[0.75±0.21\]% vs \[1.65±0.71\]%, P＝0.03) were more pronounced at 25.0 μmol/L than at 12.5 μmol/L. Conclusion: Cur is able to inhibit the growth of ARH-77 cells through activating the extrinsic apoptosis pathway and thereby may offer a potential therapeutic agent for multiple myeloma.
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Project supported by the Medical Science and Technology Project of Guangzhou （No. 20121A011102）
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