EZH2在人脂肪肉瘤的表达情况及EZH2抑制剂抗人脂肪肉瘤效果及机制研究  

Study on Expression of EZH2 in Human Liposarcoma and the Effect and Mechanism of EZH2 Inhibitor on Human Liposarcoma

作　　者：武晓慧[1,2] 徐玉乔 张东蕊 杨帆 Rajiv Kumar Jha WU Xiao-hui;XU Yu-qiao;ZHANG Dong-rui;YANG Fan;Rajiv Kumar Jha(Clinical Medical College,Xi'an Medical University,Xi'an,Shaanxi,710021,China;Department of Pathology,First Affiliated Hospital,Air Force Medical University,Xi'an,Shaanxi,710032,China;School of medical technology,Xi'an Medical University,Xi'an,Shaanxi,710021,China;China-Nepal Friendship Medical Research Center of Prof Rajiv Kumar Jha,Xi'an Medical University,Xi'an,Shaanxi,710021,China)

机构地区：[1]西安医学院临床医学院,陕西西安710021 [2]空军军医大学第一附属医院病理科,陕西西安710032 [3]西安医学院医学技术学院,陕西西安710021 [4]西安医学院中尼友好拉吉姆实验室,陕西西安710021

出　　处：《现代生物医学进展》2021年第8期1414-1419,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81550110256);西安医学院中尼友好拉吉姆医学实验室开放基金项目(18LJM02);西安医学院2019年省级大学生创新创业训练计划项目(S201911840032);西安医学院2019年校级大学生创新创业训练计划项目(121519056);西安医学院博士科研启动基金项目(2020DOC04)。

摘　　要：目的:探索组蛋白H3K27me3甲基转移酶EZH2在人脂肪瘤、高分化脂肪肉瘤和去分化脂肪肉瘤的表达情况及EZH2酶活性小分子抑制剂GSK126对人脂肪肉瘤细胞系SW872的影响,并初步探索其可能机制。方法:筛选脂肪瘤、高分化脂肪肉瘤和去分化脂肪肉瘤患者术后活检标本共计23例,其中脂肪瘤7例,高分化脂肪肉瘤9例,去分化脂肪肉瘤7例,制成组织芯片,免疫组化染色检测EZH2的蛋白表达情况。体外培养SW872脂肪肉瘤细胞系,采用CCK-8法检测不同浓度GSK126对细胞生存的抑制作用,流式细胞技术检测细胞凋亡情况,Realtime PCR法检测细胞的凋亡与抗凋亡(Caspase-1、Caspase-3、Caspase-7、Caspase-9、Bcl-2、Bag-3)、血管生成(VEGF-α)、干性(CD133、CD44、CD24)相关基因的表达,Western blot检测内质网应激相关蛋白Bip和ATF4蛋白的表达量。结果:EZH2在去分化脂肪肉瘤的表达高于高分化脂肪肉瘤,在良性脂肪瘤中的阳性表达少见(均P<0.05)。EZH2酶活性抑制剂GSK126对SW872细胞的存活有明显的抑制作用,给药后细胞凋亡率增加(P<0.05),凋亡相关基因Caspase-1、Caspase-3、Caspase-7、Caspase-9均表达增强(均P<0.05),血管生成基因VEGF-α表达降低(P<0.01),干性基因CD133表达降低(P<0.01),其余基因表达无明显差别。GSK126组的内质网应激相关蛋白Bip和ATF4蛋白的表达增加。结论:EZH2蛋白表达量与脂肪肉瘤细胞分化程度呈负相关,EZH2有望成为脂肪肉瘤的生物学标志物及恶性程度标志物。EZH2抑制剂可能成为脂肪肉瘤潜在的化疗药物,可能通过增强内质网应激发挥作用。Objective: To investigate the expression of histone H3K27me3 methyltransferase EZH2 in human liposarcoma, well differentiated liposarcoma and dedifferentiated liposarcoma, and the effect of GSK126, an enzymatic activity inhibitor of EZH2, on human liposarcoma cell line SW872, and to explore the possible mechanism. Methods: A total of 23 biopsy specimens of patients with lipoma, well differentiated liposarcoma and dedifferentiated liposarcoma were selected, including 7 cases of lipoma, 9 cases of well differentiated liposarcoma and 7 cases of dedifferentiated liposarcoma. Tissue microarray was made and the expression of EZH2 protein was detected by immunohistochemistry. SW872 liposarcoma cell line was cultured in vitro. CCK-8 method was used to detect the inhibitory effect of different concentrations of GSK126 on cell survival. The apoptosis was detected by flow cytometry. The expressions of apoptosis and anti-apoptosis(Caspase-1, Caspase-3, Caspase-7, Caspase-9, Bcl-2, Bag-3), angiogenesis(VEGF-α), stemness(CD133, CD44, CD24)genes were detected by realtime RT-qPCR, and the expression of endoplasmic reticulum stress-related proteins Bip and ATF4 were de tected by Western blot. Results: The expression of EZH2 in dedifferentiated liposarcoma was higher than that in well differentiated liposarcoma, and the positive expression in benign lipoma was rare(all P<0.05). The apoptosis rate increased(P<0.05), the expression of apoptosis related genes Caspase-1, Caspase-3, caspase-7 and Caspase-9 increased(all P<0.05), the expression of VEGF-α was decreased(P<0.05), the expression of CD133 was decreased(P<0.05). The expression of endoplasmic reticulum stress-related proteins Bip and ATF4 increased in GSK126 group. Conclusion: The expression of EZH2 protein is negatively correlated with the differentiation of liposarcoma cells, which is expected to be a biomarker of liposarcoma and malignant degree. EZH2 inhibitors may be potential chemotherapeutic agents for liposarcoma, which may play a role by enhancing endoplasmic reticu

关 键 词：EZH2 H3K27me3 脂肪肉瘤 去分化 

分 类 号：R-33[医药卫生] R739.9