Abstract
Purpose: To study whether B7-H3 and B7-H4 proteins can be candidate drug targets for preventing and treating hepatoma.Methods: Western blot assay was used to study the expressions of B7-H3 and B7-H4 proteins and the effect of sorafenib on their expressions in different human hepatoma cell lines (HepG2, Hep3B, BEL- 7402, BEL-7404, BEL-7405, QGY-7701, QGY-7703, SMMC-7721, MHCC97H, MHCC97L, HCCLM3 and HCCLM6). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to study the cytotoxicity of sorafenib against different human hepatoma cell lines.Results: The expressions of B7-H3 (0.26 - 0.84 μM) and B7-H4 (0.18 - 0.78 μM) proteins in different human hepatoma cell lines were significantly (p < 0.01) up-regulated, compared with that of the normal human liver cell line (HL-7702) (0.09 and 0.08 μM). Sorafenib was cytotoxic on Hep3B, BEL-7404, MHCC97H, HCCLM3 and HCCLM6 cells with half-maximal inhibitory concentration (IC50) of 14.56, 9.14, 9.46, 17.21 and 9.29 μM, respectively. After treatment with sorafenib at concentrations of 5, 10 and 20 μM, the expressions of B7-H3 protein in MHCC97H, HCCLM3 and HCCLM6 cells and the expressions of B7-H4 protein in Hep3B, BEL-7404 and MHCC97H cells were significantly (p < 0.01) down-regulated, compared with that of the control.Conclusion: Over-expression of B7-H3 and B7-H4 proteins is common in different human hepatoma cell lines and thus, B7-H3 and B7-H4 proteins may be regarded as candidate drug targets for preventing and treating hepatoma.Keywords: Hepatoma, B7-H3, B7-H4, Sorafenib, Over-expression, Cytotoxic activity, Down-regulation
Highlights
Primary hepatoma is the 15th most common cancer and the 2nd leading cause of cancer mortality worldwide [1]
Diphenyltetrazolium bromide (MTT) assay was used to study the cytotoxicity of sorafenib against these cell lines (Hep3B, BEL-7404, MHCC97H, HCCLM3 and HCCLM6), which had the higher expressions of B7-H3 and B7-H4 proteins than other cell lines
Western blot was used to study the expressions of B7-H3 and B7-H4 proteins in HepG2, Hep3B, BEL-7402, BEL-7404, BEL-7405, QGY-7701, QGY-7703, SMMC-7721, MHCC97H, MHCC97L, HCCLM3, HCCLM6 and HL-7702 cell lines
Summary
Primary hepatoma is the 15th most common cancer and the 2nd leading cause of cancer mortality worldwide [1]. Western blot was used to study the expressions of B7-H3 and B7-H4 proteins in different human hepatoma cell lines MTT assay was used to evaluate the antiproliferative activities of sorafenib against Hep3B, BEL-7404, MHCC97H, HCCLM3 and HCCLM6 cells. Different human hepatoma cells were separately seeded on 96 well plates for 24 h, and cells were treated with sorafenib at the concentrations of 0 (for control), 1.25, 2.5, 5, 10, 15, 20 and 40 μM. The total proteins of untreated HepG2, Hep3B, BEL-7402, BEL-7404, BEL-7405, QGY-7701, QGY-7703, SMMC-7721, MHCC97H, MHCC97L, HCCLM3, HCCLM6 and HL-7702 cells were extracted. After treatment with sorafenib at concentrations of 0 (for control), 5, 10 and 20 μM for 48 h, the total proteins of Hep3B, BEL-7404, MHCC97H, HCCLM3 and HCCLM6 were extracted. Trop J Pharm Res, October 2015; 14(10): 1798 was less than 0.05, the differences were recognized as statistically significant
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