Effect and Mechanism of Celastrol on the Renal Clear Cell Carcinoma 786-0 Cells

Abstract

Background The incidence rate of renal clear cell carcinoma was high and the cure rate was low. Celastrol could regulate cytokines in tumor cells, then inhibit the proliferation of tumor cells. However, there were few studies on the effect of celastrol on the renal clear cell carcinoma 786-0 cells. Objective To explore the effect and mechanism of celastrol on the renal clear cell carcinoma 786-0 cells, in order to provide a reference for investigating drugs for the treatment of renal clear cell carcinoma. Methods From February 2015 to May 2016, renal clear cell carcinoma 786-0 cells and normal renal tubular epithelial cells HK2 cells (renal cells HK2 cells) were cultured. The survival rate of renal clear cell carcinoma 786-0 cells, renal cells HK2 cells were detected after treating with celastrol at different concentrations (0, 0.5, 1.0, 1.5, 2.0 μmol/L) for different time(12, 24, 48 h); cytotoxicity of renal clear cell carcinoma 786-0 cells, renal cells HK2 cells were detected after treating with celastrol at different concentrations (0, 0.5, 1.0, 1.5, 2.0 μmol/L),1.5 μmol/L 5-Fu; cell cycle, apoptosis rate of renal clear cell carcinoma 786-0 cells, renal cells HK2 cells were detected after treating with celastrol at different concentrations (0, 1.5, 2.0 μmol/L), 1.5 μmol/L 5-Fu; mitochondrial membrane potential of renal clear cell carcinoma 786-0 cells, renal cells HK2 cells were measured after treating with celastrol at different concentrations (0, 0.5, 1.0, 1.5 μmol/L); cysteine-containing aspartic proteolytic enzymes (caspase)-3, caspase-9 activity, the levels of Bax, B lymphocyte tumor-2(Bcl-2), cytochrome c and polyribosylribitol phosphate (PRRP) in the renal clear cell carcinoma 786-0 cells were detected after treating with celastrol at different concentrations (0, 0.5, 1.0, 1.5, 2.0 μmol/L). Results The survival rate of the renal cell carcinoma 786-0 cells decreased with the increase of concentration when treated with 0, 0.5, 1.0, 1.5, 2.0 μmol/L celastrol for 12, 24, 48 h (P<0.05). The survival rate of the renal cell carcinoma 786-0 cells treated with 1.0 μmol/L celastrol for 24, 48 h were less than that for 12 h (P<0.05); the survival rate of the renal cell carcinoma 786-0 cells treated with 1.5, 2.0 μmol/L celastrol for 12, 24, 48 h decreased with the increase of time (P<0.05). Cytotoxicity of renal clear cell carcinoma 786-0 cells, renal cells HK2 cells treated with 0.5 μmol/L, 1.0 μmol/L, 1.5 μmol/L, 2.0 μmol/L celastrol and 1.5 μmol/L 5-Fu were larger than 0 μmol/L celastrol (P<0.05); cytotoxicity of renal clear cell carcinoma 786-0 cells treated with 2.0 μmol/L celastrol was larger than 0.5, 1.0, 1.5 μmol/L celastrol (P<0.05); cytotoxicity of renal cells HK2 cells treated with 2.0 μmol/L celastrol was larger than 1.0 μmol/L celastrol (P<0.05); cytotoxicity of renal cell carcinoma 786-0 cells and renal cells HK2 cells treated with 1.5 mmol/L 5-Fu was larger than 0.5, 1.0, 1.5, 2.0 μmol/L celastrol (P<0.05). After treated with 0, 1.5 mmol/L celastrol, the renal cell carcinoma 786-0 cells and renal cells HK2 cells at the G2/M phase were less than S, G0/G1 phase (P<0.05); after treated with 2.0 umol/L celastrol, 1.5 umol/L 5-Fu, renal clear cell carcinoma 786-0 cells, renal cells HK2 cells at G0/G1 phase were more than S phase, while renal clear cell carcinoma 786-0 cells and renal cells HK2 cells at G2/M phase were less than S, G0/G1 phase (P<0.05). Cell apoptosis rate of renal clear cell carcinoma 786-0 cells treated with 1.5 μmol/L, 2.0 μmol/L celastrol and 1.5 μmol/L 5-Fu were larger than 0 μmol/L celastrol (P<0.05); cell apoptosis rate of renal clear cell carcinoma 786-0 cells treated with 2.0 μmol/L celastrol and 1.5 μmol/L 5-Fu were larger than 1.5 μmol/L celastrol (P<0.05); cell apoptosis rate of renal cells HK2 cells treated with 1.5 μmol/L 5-Fu were larger than 0 μmol/L, 2.0 μmol/L celastrol (P<0.05). Mitochondrial potential of renal clear cell carcinoma 786-0 cells treated with 0, 0.5, 1.0, 1.5 μmol/L celastrol increased in turn (P<0.05). caspase-3, caspase-9 activity of renal clear cell carcinoma 786-0 cells treated with 0, 0.5, 1.0, 1.5 μmol/L celastrol increased in turn (P<0.05). The levels of Bax, cytochrome c, PRRP of renal clear cell carcinoma 786-0 cells treated with 0, 0.5, 1.0, 1.5 μmol/L celastrol increased in turn, while the level of Bcl-2 decreased in turn (P<0.05). Conclusion Celastrol may inhibit the proliferation of renal clear cell carcinoma 786-0 cells and induce their apoptosis by activating caspase-3, caspase-9 activity, upregulating Bax, cytochrome c, PRRP levels and down-regulating Bcl-2 level.