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目的探讨沉默信息调节因子2-相关酶3(SIRT 3)对肾透明细胞癌细胞系786-O中锰型超氧化物歧化酶(MnSOD)的去乙酰化作用及其对786-O细胞增殖、凋亡的影响。方法Western blotting和免疫共沉淀(IP)测定786-O中MnSOD乙酰化的水平;MnSOD酶活性试剂盒、四甲基偶氮唑盐(MTT)和Hoechst荧光染色分别检测肾癌细胞中MnSOD酶活性、细胞增殖和凋亡。结果与空白载体(pcDNA3.0)相比,SIRT 3(转染pcDNA3.0-sirt3)使MnSOD乙酰化水平由1.29±0.16降低为0.74±0.07(t=7.21,P<0.001),其酶活性由(1.47±0.17)U/mg增加至(2.53±0.31)U/mg(t=6.70,P<0.001)、细胞增殖率由(25.28±5.75)%增加至(48.86±7.47)%(t=5.60,P<0.001),而细胞凋亡率由(4.53±0.51)%变为(4.45±0.59)%,差异无统计学意义(t=0.24,P=0.82)。结论肾透明细胞癌786-O中MnSOD的去乙酰化增强其细胞增殖能力,MnSOD或其乙酰化可能是肾透明细胞癌治疗的一个潜在靶点。
Objective To investigate the deacetylation of SIRT3 on MnSOD in 786-O cell line and its effect on the proliferation, Effect of apoptosis. Methods The levels of MnSOD acetylation in 786-O cells were determined by Western blotting and immunoprecipitation (IP). The activity of MnSOD in renal cell carcinoma cells was detected by MnSOD enzyme activity assay, MTT assay and Hoechst staining. , Cell proliferation and apoptosis. Results SIRT 3 (transfected with pcDNA3.0-sirt3) reduced the level of MnSOD acetylation from 1.29 ± 0.16 to 0.74 ± 0.07 (t = 7.21, P <0.001) compared with blank vector (pcDNA3.0) The cell proliferation rate increased from (25.28 ± 5.75)% to (48.86 ± 7.47)% (t = 0.65)% from 1.47 ± 0.17 U / mg to 2.53 ± 0.31 U / 5.60, P <0.001), while the apoptotic rate changed from (4.53 ± 0.51)% to (4.45 ± 0.59)%, with no significant difference (t = 0.24, P = 0.82). Conclusion Deacetylation of MnSOD in renal clear cell carcinoma 786-O enhances cell proliferation. MnSOD or acetylation may be a potential target for the treatment of renal clear cell carcinoma.