灵芝烯酸B诱导Oct4去SUMO化激活G0期胃癌侧群细胞  

Ganoderma acid B induces Oct4 deSUMOylation and activates gastric cancer side population cells in G0 phase

作　　者：成睿珍 张春艳 刘凤婷 李睿 吴文静 李艳霞[3] 马晓芳[3] 李丽丽 刘晓智[3] CHENG Ruizhen;ZHANG Chunyan;LIU Fengting;LI Rui;WU Wenjing;LI Yanxia;MA Xiaofang;LI Lili;LIU Xiaozhi(Department of Pharmacy,Tianjin Binhai New Area Hospital of Traditional Chinese Medicine,Tianjin 300450,China;The College of Clinical Medicine of Tianjin Medical University,Tianjin 300070,China;The Central Laboratory,The Fifth Central Hospital of Tianjin,Tianjin 300450,China;Department of Bone and Soft Tissue Oncology,Tianjin Medical University Cancer Institute and Hospital,National Clinical Research Center for Cancer,Key Laboratory of “Cancer Prevention and Therapy” of Tianjin,Tianjin Clinical Research Center for Malignant Tumor,Tianjin 300060,China)

机构地区：[1]天津市滨海新区中医医院药学科,天津300450 [2]天津医科大学临床医学院,天津300070 [3]天津市第五中心医院中心实验室,天津300450 [4]天津医科大学肿瘤医院骨与软组织肿瘤科国家肿瘤临床医学研究中心天津市"肿瘤防治"重点实验室天津市恶性肿瘤临床医学研究中心,天津300060

出　　处：《天津中医药》2019年第4期396-399,共4页Tianjin Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81471175)

摘　　要：[目的]从蛋白质类泛素化修饰角度研究灵芝烯酸B(GAB)对静息期G0胃癌侧群细胞的活化作用,为胃癌侧群细胞靶向治疗提供新策略。[方法]流式细胞术分离CD133+的SGC-7901胃癌侧群细胞,培养细胞克隆球,给予GAB处理96 h,于不同时间点检测克隆球大小,绘制克隆球变化曲线;流式细胞术检测胃癌侧群细胞周期变化;蛋白免疫印迹(Western Blot)方法检测小泛素样修饰蛋白-1(SUMO-1)、八聚体结合转录因子4(Oct4)、细胞周期蛋白Cyclin D1、Cyclin E1、Cyclin A2和Cyclin B1的表达水平;四甲基噻唑蓝(MTT)法检测顺铂对CD133+胃癌侧群细胞的半数致死量(IC50)。[结果] GAB组胃癌侧群细胞克隆球生长速度明显低于对照组;对照组CD133+胃癌侧群细胞主要集中于G0/G1期,而GAB组CD133+则主要集中于G2/M期;与对照组比较,GAB组胃癌侧群细胞的SUMO-1、Oct4和Cyclin D1蛋白表达水平降低,而Cyclin E1、Cyclin A2和Cyclin B1蛋白表达水平升高(P<0.05);GAB处理使顺铂对胃癌侧群细胞IC50从69.84μg/mL下降至35.67μg/mL。[结论] GAB能够通过诱导Oct4去SUMO化激活G0期胃癌侧群细胞,增加胃癌侧群细胞对顺铂的化疗敏感性。[Objective] To study the activation effect of Ganoderma acid B (GAB) on gastric cancer side population cells in resting stage from the perspective of protein SUMOylation,and to provide a new strategy for the targeted therapy of gastric cancer side population cells.[Methods] Flow cytometry was used to isolate the SGC-7901 gastric cancer side population cells,and then treated with the active ingredient of traditional Chinese medicine,Ganoderma acid B (GAB) with final concentration of 10 μmol/L for 96 h. The diameter of cell clone was plotted at different time points. The change of cell cycle of gastric cancer side population cells was detected by flow cytometry technique. The expression level of small ubiquitin-like modifier (SUMO)-1,eight octamer binding transcription factors 4 (Oct4),Cyclin D1,Cyclin E1,Cyclin A2 and Cyclin B1 protein was detected by Western Blot assay. The median lethal dose (IC50) of cisplatin on gastric cancer side population cells was detected by MTT assay.[Results] The growth rate of gastric cancer side population cell clone balls in GAB group was significantly slower than the control group. The CD133+ gastric cancer side population cells in control group mainly in G0/G1 phase,and the GAB group cells mainly in G2/M phase. Compared with the control group,the expression level of Oct4, SUMO-1 and Cyclin D1 proteins was significantly decreased,but the level of Cyclin E1,Cyclin A2 and Cyclin B1 proteins was increased in GAB group (P<0.05). GAB induced IC50 from 69.84 g/mL down to 35.67 g/mL in gastric cancer side population cells to cisplatin.[Conclusion] GAB can activate resting gastric cancer side population cells by inducing Oct4 deSUMOylation,and increase their sensitivity to cisplatin.

关 键 词：胃癌侧群细胞 小泛素样修饰蛋白 八聚体结合转录因子4 顺铂 灵芝烯酸B 

分 类 号：R285.5[医药卫生—中药学]