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目的 探讨沉默信息调节因子 ２-相关酶 ３（ＳＩＲＴ３）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 Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ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，ＳＩＲＴ３（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 effect of SIRT3 on deacetylation of manganese-type superoxide dismutase (MnSOD) and its effect on the proliferation and apoptosis of 786-O cells in 786-O renal clear cell carcinoma cell line 786-O The impact of death. 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 Compared with blank vector (pcDNA3.0), SIRT3 (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). The enzyme activity increased from 1.47 ± 0.17 U / mg to 2.53 ± 0.31 U / mg (t = 6.70, P <0.001) ), The proliferation rate increased from (25.28 ± 5.75)% to (48.86 ± 7.47)% (t = 5.60, P <0.001) .53 ± 0.51)% (4.45 ± 0.59)%, the difference was not statistically significant (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.