MnSOD乙酰化对肾透明细胞癌786-O细胞增殖、凋亡的影响

发布时间：2018-12-12 01:13

【摘要】：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,P0.001),其酶活性由(1.47±0.17)U/mg增加至(2.53±0.31)U/mg(t=6.70,P0.001)、细胞增殖率由(25.28±5.75)%增加至(48.86±7.47)%(t=5.60,P0.001),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。
[Abstract]:Objective to investigate the deacetylation of manganese-type superoxide dismutase (MnSOD) in renal clear cell carcinoma cell line 786-O by silencing information regulatory factor 2-associated enzyme 3 (SIRT 3) and its effect on proliferation and apoptosis of 786-O cell line. Methods Western blotting and co-immunoprecipitation (IP) were used to detect the level of MnSOD acetylation in 786-O cells, and MnSOD enzyme activity kit, (MTT) and Hoechst fluorescence staining were used to detect MnSOD activity, cell proliferation and apoptosis, respectively. Results compared with blank vector (pcDNA3.0), SIRT 3 (transfection of pcDNA3.0-sirt3) reduced the acetylation level of MnSOD from 1.29 卤0.16 to 0.74 卤0.07 (T7.21mP0.001). The enzyme activity increased from (1.47 卤0.17) U/mg to (2.53 卤0.31) U/mg (tchr 6.70) P0.001, and the cell proliferation rate increased from (25.28 卤5.75)% to (48.86 卤7.47)% (t 5.60 P0.001). However, the rate of apoptosis changed from (4.53 卤0.51)% to (4.45 卤0.59)%, and there was no significant difference between them. Conclusion the deacetylation of MnSOD in renal clear cell carcinoma 786-O enhances its proliferative ability. MnSOD or acetylation may be a potential target for the treatment of renal clear cell carcinoma.
【作者单位】：山东大学附属千佛山医院小儿外科;山东大学齐鲁医学部基础医学院癌症研究中心;山东大学附属省立医院泌尿外科;
【基金】：山东省科技重大专项(2015ZDXX0802A02) 山东省医药卫生科技发展计划(2016WS0481)
【分类号】：R737.11

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