蛋白酶体抑制剂RA190对口腔腺样囊性癌的调节作用  

Regulatory effect of the proteasome inhibitor RA190 on oral adenoid cystic carcinoma

作　　者：赵曦[1] 郑黎薇[2] 周丹[3] 王春 Xi Zhao;Li-wei Zheng;Dan Zhou;Chun Wang(Department of stomatology,People's Hospital of Deyang City,Deyang,Sichuan 618000,China;The State Key Laboratory of Oral Disease&National Clinical Research Center for Oral Diseases,West China Hospital of Stomatology,Sichuan University,Chengdu,Sichuan 610041,China;Department of Anesthesiology,People's Hospital of Deyang City,Deyang,Sichuan 618000,China)

机构地区：[1]德阳市人民医院口腔科,四川德阳618000 [2]四川大学华西口腔医院(口腔疾病研究国家重点实验室),四川成都610041 [3]德阳市人民医院麻醉科,四川德阳618000

出　　处：《中国现代医学杂志》2022年第15期53-58,共6页China Journal of Modern Medicine

基　　金：国家自然科学基金(No:81771033)。

摘　　要：目的探讨蛋白酶体抑制剂双苄基吡啶酮(RA190)通过非ATP酶依赖性调节颗粒13(Rpn13)抑制泛素-蛋白酶体通路途径对口腔腺样囊性癌的作用,揭示RA190对腺样囊性癌ACC-2细胞增殖、凋亡的调节作用和可能机制。方法确定ACC-2细胞表达Rpn13后,选择RA190对ACC-2细胞的合适剂量(600 nmol/L);实验分为ACC-2组、ACC-2-Rpn13-siRNA组、RA190-ACC-2-Rpn13-siRNA组;敲除ACC-2细胞中Rpn13的表达;用RA190处理ACC-2细胞;用RA190处理敲除Rpn13的ACC-2细胞;MTT法观察各组细胞增殖、凋亡情况;Western blotting检测各组细胞Survivin、TIP30、泛素化蛋白相对表达量;qRT-PCR检测Survivin和TIP30 mRNA相对表达量。结果敲除Rpn13后的细胞组中Rpn13蛋白相对表达量较ACC-2细胞组降低(P<0.05)。MTT法检测显示,不同剂量的RA190作用ACC-2细胞后光密度(OD)比较,差异有统计学意义(P<0.05)。与ACC-2组比较,其余3组OD值均降低(P<0.05)。与ACC-2组比较,其余3组Survivin蛋白相对表达量均降低(P<0.05),TIP30蛋白相对表达量均升高(P<0.05),RA190-ACC-2组TIP30蛋白相对表达量最高,泛素化蛋白聚集最明显(P<0.05)。与ACC-2组比较,其余3组Survivin mRNA的相对表达量均降低(P<0.05),TIP30 mRNA相对表达量均升高(P<0.05)。结论RA190促凋亡的活动主要取决于Rpn13,Rpn13是口腔腺样囊性癌的致癌基因。RA190的调节作用可能改善口腔腺样囊性癌的预后,Rpn13可能成为治疗口腔腺样囊性癌的靶点。Objective To investigate the effect of the proteasome inhibitor dibenzylpyridone RA190 on ACC-2(oral adenoid cystic carcinoma)by inhibiting UPP(ubiquitin-26S proteasome pathway)via Rpn13(non ATP-dependent regulatory granule 13),and thus revealing its effect on ACC-2 cell proliferation,the regulatory role,and possible mechanism of apoptosis.Methods After determining the expression of Rpn13 in ACC-2 cells,the appropriate dose of RA190 for ACC-2 cells were selected.Finally,the expression of survivin,TIP30,ubiquitinated protein of four groups(A:ACC-2 cell group;group B:ACC-2-Rpn13-siRNA(small interfering RNA)group knockdown the expression of Rpn13 in ACC-2 cells;group C:group RA190-ACC-2 group,treated A group with RA190;group D:RA190-AC2-Rpn13-SiRNA treated B group with RA190)were detected.The proliferation and apoptosis of each group were observed by MTT.Results Compared to the ACC-2 group,the OD Values of another 3 groups were decreased;compared to the ACC-2 group,the relative expression of Survivin was reduced,and the relative expression of TIP30 was increased in the RA190-ACC-2 group;compared to the ACC-2-Rpn13-siRNA group,in the RA190-ACC-2-Rpn13-siRNA group,the relative expression of TIP30 was reduced,and the relative expression of Survivin was increased;the aggregation of ubiquitinated protein in the RA190-ACC-2 group was the most obviously;the Survivin mRNA valus in another 3 groups were lower than the ACC-2 group,and the TIP30 mRNA valus in another 3 groups were higher than the ACC-2 group.Above all,the differences were significantly(all P<0.05).Conclusion RA190 promoted ACC-2 cell apoptosis and inhibited proliferation through Rpn13,may improve the prognosis of oral adenoid cystic carcinoma,and can be used as a target for the treatment of oral adenoid cystic carcinoma.

关 键 词：口腔腺样囊性癌 蛋白酶体抑制剂 双苄基吡啶酮 非ATP酶依赖性调节颗粒13 泛素-蛋白酶体通路 凋亡 

分 类 号：R739.8[医药卫生—肿瘤]