SUMO特异性蛋白酶1诱导蛋白质去小泛素相关修饰增加子宫内膜癌侧群细胞化疗敏感性  

SENP1 induced protein deSUMO modification increased the chemotherapy sensitivity of endometrial cancer side population cells

作　　者：袁梦岚[1] 白洁 李承尧 薛娜 陈旭红[1] 盛凤[4] 刘晓智 李圃[2] Yuan Menglan;Bai Jie;Li Chengyao;Xue Na;Chen Xuhong;Sheng Feng;Liu Xiaozhi;Li Pu(Department of Obstetrics and Gynecology,Tianjin Fifth Central Hospital,Tianjin 300450,China;Department of Obstetrics and Gynecology,Tianjin Central Hospital of Gynecology Obstetrics,Tianjin 300052,China;Tianjin Key Laboratory of Epigenetics in Organ Development of Premature Infants,Tianjin 300450,China;Department of Traditional Chinese Medicine,Tianjin Fifth Central Hospital,Tianjin 300450,China)

机构地区：[1]天津市第五中心医院妇产科,天津300450 [2]天津市中心妇产科医院妇产科,天津300052 [3]天津市早产儿器官发育表观遗传重点实验室,天津300450 [4]天津市第五中心医院中医科,天津300450

出　　处：《中华肿瘤杂志》2022年第12期1362-1368,共7页Chinese Journal of Oncology

基　　金：天津市新一代人工智能科技重大专项项目(18ZXZNSY00260);天津市滨海新区卫生和计划生育委员会科技项目(2018BWKY013、ZC20140)。

摘　　要：目的探讨通过诱导蛋白质广泛去类泛素化修饰途径抑制子宫内膜癌的干性维持潜能,达到子宫内膜癌侧群细胞化疗增敏的目的。方法流式细胞术分选培养CD133^(+)CD44^(+)的KLE子宫内膜癌侧群细胞克隆球,Western blot法检测小泛素相关修饰蛋白1(SUMO1)及肿瘤侧群细胞干性维持基因八聚体结合转录因子4(Oct4)和性别决定区Y-box2(Sox2)蛋白的表达情况。慢病毒介导KLE侧群细胞稳定转染SUMO特异性蛋白酶1(SENP1)基因,Western blot法检测SENP1、SUMO1、Oct4和Sox2的蛋白表达;比较过表达与未过表达SENP1基因的KLE侧群细胞克隆形成率,流式细胞术检测细胞周期变化,四甲基偶氮唑蓝实验和流式细胞术检测子宫内膜癌侧群细胞对顺铂的化疗敏感性,子宫内膜癌荷瘤鼠模型检测SENP1过表达对顺铂的化疗敏感性影响。结果CD133^(+)CD44^(+)和CD133-CD44-子宫内膜癌KLE细胞克隆形成率分别为(23.64±5.03)%和(4.64±1.25)%,差异有统计学意义(P=0.003),CD133^(+)CD44^(+)子宫内膜癌KLE细胞中SUMO1、Oct4和Sox2蛋白表达水平高于CD133-CD44-子宫内膜癌KLE细胞(均P<0.05)。与未转染SENP1基因的KLE侧群细胞比较,SENP1过表达的KLE侧群细胞SUMO1、Oct4、Sox2蛋白表达水平降低,细胞克隆形成率由(25.67±5.44)%下降至(7.46±1.42)%,细胞周期发生由G0/G_(1)期向G_(2)期的偏移,顺铂半数抑制浓度由(55.46±6.14)μg/ml下降至(11.55±3.12)μg/ml,细胞凋亡率由(9.76±2.09)%上升至(16.79±3.44)%,差异均有统计学意义(均P<0.05)。过表达SENP1能够降低KLE侧群细胞增殖能力,增加化疗敏感性。结论通过过表达SENP1能够诱导蛋白质去SUMO化修饰,抑制子宫内膜癌侧群细胞的干性维持潜能,进而增强其化疗敏感性。Objective To inhibit the stemness maintenance potential of endometrial cancer and increase the sensitivity of endometrial cancer side population cells to chemotherapy drugs by inducing extensive deSUMOylation modification of proteins.Methods Flow cytometry was used to sort and culture CD133^(+)CD44^(+)KLE endometrial cancer cell clone spheres.Protein expression level of small ubiquitin-related modifier 1(SUMO1)and two stemness maintenance genes of tumor side population cells,octamer binding transcription factor-4(Oct4)and sex determining region Y-box2(Sox2),were detected by western blotting method.Lentivirus-mediated Sentrin/SUMO-specific proteases 1(SENP1)gene was stably transfected into KLE side population cells.Western blotting was used to detect the protein expressions of SENP1,SUMO1,Oct4 and Sox2.The clone formation rate was compared between KLE side population cells with or without SENP1 overexpression.Flow cytometry was applied to detect cell cycle changes.3-(4,5-Dimethylthiazole-2)-2,5-diphenyl-tetrazolium bromide(MTT)experiment and flow cytometry apoptosis method were used to detect the chemosensitivity of the side population of endometrial cancer cells to cisplatin.Tumor-bearing mouse models of endometrial cancer were established to detect the effect of SENP1 overexpression on the chemotherapy sensitivity of cisplatin.Results Compared with CD133-CD44-KLE cells,CD133^(+)CD44^(+)KLE side population cells could form clonal spheres and express higher levels of SUMO1,Oct4 and Sox2 proteins(P<0.05).Compared with KLE side population cells that were not transfected with SENP1 gene,the expression level of SENP1 protein in KLE side population cells overexpressing SUMO1、Oct4 and Sox2 were lower.The clonal sphere formation rate was reduced from(25.67±5.44)%to(7.46±1.42)%,and cell cycle shifted from G0/G_(1) phase to G_(2) phase.IC50 of cisplatin decreased from(55.46±6.14)μg/ml to(11.55±3.12)μg/ml,and cell apoptosis rate increased from(9.76±2.09)%to(16.79±3.44)%.Overexpression of SENP1 could reduce the tum

关 键 词：子宫内膜肿瘤 小泛素相关修饰蛋白 SUMO特异性蛋白酶1 肿瘤侧群细胞 顺铂 

分 类 号：R737.33[医药卫生—肿瘤]