[目的]探讨异柠檬酸脱氢酶(IDH)132、140、172位精氨酸突变对骨巨细胞瘤细胞增殖水平的影响.[方法]纳入2020年1月-2022年1月骨巨细胞瘤患者76例,检测骨巨细胞瘤组织和血清中IDH1和IDH2的基因序列和IDH底物2-羟基戊二酸(2HG)的水平.构建IDH1和IDH2敲除的骨巨细胞瘤细胞GCT-404细胞系,转染IDH野生型或突变型后检测细胞的增值水平.根据转染的IDH1或IDH2的基因型进行实验分组:①IDH1野生型组;②IDH2野生型组;③IDH1 R132H组;④IDH2 R140Q组;⑤IDH2 R172S组.[结果]骨巨细胞瘤患者中IDH突变型占比32.89％,野生型占比67.11％.IDH野生型患者肿瘤组织和血清中2HG的水平低于IDH突变型患者[(0.22±0.07)nmol/mg vs(2.58±0.24)nmol/mg,(1.73±0.29)µmol/L vs(9.89±1.08)μmol/L,P＜0.05].相比于 IDH 野生型,IDH1 R132H、IDH2 R140Q 或 IDH2 R172S 显著促进 GCT-404 细胞的增殖(2.20±0.09 vs 4.25±0.12,P＜0.05)和 2HG 水平(31.34±5.33 vs 56.32±8.32,P＜0.05).敲除IDH1 或 IDH2 后,GCT-404 细胞中 2HG 的水平显著下降(25.78±4.56 vs 10.22±2.04,P＜0.05).2HG 处理后,GCT-404 细胞的增殖水平均显著上升(1.56±0.08 vs 2.41±0.10,P＜0.05).相比于IDH野生型,IDH1 R132H、IDH2 R140Q或IDH2 R172S显著增加GCT-404细胞中HIF1A的表达.敲低 HIF1A 后 GCT-404 细胞的增殖水平显著下降(1.50±0.04 vs 0.94±0.03,P＜0.05).[结论]IDH1 R132H、IDH2 R140Q或IDH2 R172S通过调控2HG水平和HIF1A水平能显著增强骨巨细胞瘤细胞的增殖水平.
Effect of isocitrate dehydrogenase mutation on proliferation of giant cell tumor of bone
[Objective]To investigate the effects of arginine mutations at positions 132,140 and 172 of isocitrate dehydrogen-ase(IDH)on the proliferation of bone giant cell tumor cells.[Method]Seventy-six patients with giant cell tumor of bone ad-mitted to our hospital from January 2020 to January 2022 were included.The gene sequences of IDH1 and IDH2 and the level of IDH substrate 2-hydroxy-glutaric acid(2HG)in the tissues and serum of giant cell tumor of bone were detected.GCT-404 cell lines of bone giant cell tumor knocked out with IDH1 and IDH2 were constructed,and the proliferation level of the cells was detected after transfection with wild type or mutant type of IDH.The transfected IDH1 or IDH2 genotypes were divided into the following groups:①IDH 1 wild type group;②IDH2 wild-type group;③IDH1 R132H group;④IDH2 R140Q group;⑤IDH2 R172S group.[Result]The mutant IDH accounted for 32.89％of the patients with giant cell tumor of bone,and the wild type accounted for 67.11％.The level of 2HG in tumor tissue and serum of IDH wild-type patients was lower than that of IDH mu-tant patients[(0.22±0.07)nmol/mg vs(2.58±0.24)nmol/mg,(1.73±0.29)μmol/L vs(9.89±1.08)µmol/L,P＜0.05].Compared with IDH wild type,IDH1 R132H,IDH2 R140Q or IDH2 R172S significantly promoted the proliferation of GCT-404 cells(2.20±0.09 vs 4.25±0.12,P＜0.05)and 2HG(31.34±5.33 vs 56.32±8.32,P＜0.05).After IDH1 or IDH2 knockout,the level of 2HG in GCT-404 cells was significantly decreased(25.78±4.56 vs 10.22±2.04,P＜0.05).After 2HG treatment,the proliferation levels of GCT-404 cells were significantly increased(1.56±0.08 vs 2.41±0.10,P＜0.05).Compared with IDH wild type,IDH1 R132H,IDH2 R140Q or IDH2 R172S significantly increased the expression of HIF1A in GDT-404 cells.The proliferation level of GCT-404 cells was significantly decreased after HIF1A knockdown(1.50±0.04 vs 0.94±0.03,P＜0.05).[Conclusion]IDH1 R132H,IDH2 R140Q or IDH2 R172S can significantly enhance the proliferation level of bone giant cell tumor by regulating the level of 2HG and HIF1A.
isocitrate dehydrogenasegiant cell tumor of boneproliferation levelHIF1A2-hydroxyglutaric acid
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