摘要
目的探讨乙酰基转移酶抑制剂NU9056对食管癌EC109细胞乙酰基转移酶KAT5、Survivin乙酰化水平及细胞增殖和迁移的影响。方法NU9056处理食管癌EC109细胞为实验组(NU9056组),有机溶剂DMSO处理食管癌EC109细胞为对照组(DMSO组)。采用MTT法筛选并确定NU905最佳给药浓度,RT-qPCR检测KAT5 mRNA表达,Western blot检测KAT5、细胞周期蛋白D1(Cyclin D1)、癌基因-Myc(c-Myc)、B淋巴细胞瘤-2(BCL2)和血管内皮生长因子(VEGF)蛋白表达,免疫共沉淀检测Survivin乙酰化水平,MTT法检测细胞增殖情况,划痕修复实验检测迁移能力,Transwell小室实验检测侵袭能力。结果NU9056可浓度依赖性抑制食管癌EC109细胞增殖,其IC_(50)=0.946μmol/L,确定NU905最佳给药浓度为0.9μmol/L。NU9056组中KAT5 mRNA和蛋白表达较DMSO组明显降低,差异有统计学意义(P<0.05)。NU9056组Survivin乙酰化水平较DMSO组明显下降,差异有统计学意义(P<0.05)。NU9056组肿瘤增殖相关蛋白Cyclin D1、c-Myc、BCL2和VEGF蛋白表达,细胞的增殖能力、侵袭能力、迁移能力较DMSO组明显降低(P<0.05)。结论NU9056可能通过抑制乙酰基转移酶KAT5的表达下调食管癌EC109细胞中Survivin乙酰化水平,进一步抑制肿瘤增殖通路中相关蛋白表达,从而抑制食管癌细胞增殖、侵袭和迁移能力。
Objective To study the effect of acetyltransferase inhibitor NU9056 on acetyltransferase KAT5,Survivin acetylation level and cellular proliferation and migration of esophageal cancer EC109 cells.Methods Esophageal cancer EC109 cells treated with NU9056 served as the experimental group(NU9056 group) and esophageal cancer EC109 cells treated with organic solvent DMSO as the control group(DMSO group).The optimal NU905 administration concentration was screened and determined by the MTT method.The relative expression level of KAT5 mRNA was measured by RT-qPCR.The expression levels of the KAT5,Cyclin D1,c-Myc, BCL2 and VEGF proteins were determined by Western blot.The acetylation level of Survivin protein was detected by immunoprecipitation, the cell proliferation was detected by MTT assay, the migration ability was determined by scratch repair assay and the invasion ability was observed in Transwell chambers.Results NU9056 concentration-dependent inhibited esophageal cancer EC109 cell proliferation, and its IC_(50)=0.946 μmol/L,the optimal administration concentration of NU9056 was determined as 0.9 μmol/L.The KAT5 mRNA and protein expression in the NU9056 group were decreased significantly compared with the DMSO group, with a statistically significant difference(P<0.05).The Survivin acetylation in the NU9056 group was significantly decreased compared with the DMSO group, and the difference was statistically(P<0.05).The expression levels of the tumor proliferation-related proteins Cyclin D1,c-Myc, BCL2 and VEGF were significantly down-regulated after NU9056 down-regulating Survivin acetylation levels in EC109 cells(P<0.05);the proliferation capacity, invasion capacity and migration capacity in the NU9056 group were significantly decreased(P<0.05).Conclusion NU9056 may down-regulate the Survivin protein acetylation level in esophageal cancer EC109 cells by inhibiting acetyltransferase KAT5 expression, further inhibit the expression of key proteins in the tumor proliferation pathway, thus inhibit the proliferation, in
作者
梁宗英
杨阳
郑竞雄
赵宝山
侯继申
孙光蕊
LIANG Zongying;YANG Yang;ZHENG Jingxiong;ZHAO Baoshan;HOU Jishen;SUN Guangrui(Department of Thoracic Surgery,Affiliated Hospital of Chengde Medical College,Chengde,Hebei 067000,China;Department of Clinical Laboratory,Hebei Provincial Chest Hospital,Shijiazhuang,Hebei 050000,China)
出处
《重庆医学》
CAS
2023年第2期167-171,共5页
Chongqing medicine
基金
河北省医学科学研究重点课题计划项目(20210985)
承德市科学技术研究与发展计划项目(201606A036)。
