敲低WDHD1对鼻咽癌细胞增殖、铜含量及铜死亡相关因子的影响  

Effect of WDHD1 knockdown on proliferation,copper content and copper death⁃related factors in nasopharyngeal carcinoma cells

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作  者:言柯柯 伍小薇 杨振东 谢小燕 周慧斐 罗嘉嫄 韦祝新[1] YAN Keke;WU Xiaowei;YANG Zhendong;XIE Xiaoyan;ZHOU Huifei;LUO Jiayuan;WEI Zhuxin(Department of Radiotherapy,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi,China;Department of Pathology,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi,China)

机构地区:[1]广西医科大学第一附属医院放疗科,广西南宁市530021 [2]广西医科大学第一附属医院病理科,广西南宁市530021

出  处:《广西医学》2024年第10期1519-1527,共9页Guangxi Medical Journal

基  金:广西自然科学基金(2024GXNSFAA010117)。

摘  要:目的探讨敲低WD重复序列和HMG框DNA结合蛋白1(WDHD1)对鼻咽癌细胞增殖、铜含量及铜死亡相关因子的影响。方法将鼻咽癌细胞(HK-1细胞和HONE-1细胞)分别分为对照组、敲低空载组(sh-NC组)、WDHD1敲低组(sh-WDHD1组)。对照组不做处理,sh-NC组、sh-WDHD1组分别转染阴性对照慢病毒、sh-WDHD1 RNA慢病毒。转染成功后,检测各组HK-1细胞和HONE-1细胞增殖活性、铜含量、铜死亡相关基因m RNA及蛋白表达情况;将sh-WDHD1组的HK-1细胞和HONE-1细胞分为sh-WDHD1对照组、sh-WDHD1/DMSO组、sh-WDHD1/ATTM组,分别使用完全培养基、含DMSO的完全培养基、含铜螯合剂ATTM的完全培养基进行培养,然后检测细胞铜含量。结果(1)与对照组和sh-NC组相比,sh-WDHD1组HONE-1细胞在24 h、48 h的增殖活力下降,HK-1细胞在48 h的增殖活力下降(P<0.05)。(2)在HONE-1细胞和HK-1细胞中,与对照组及sh-NC组相比,sh-WDHD1组的细胞铜含量增加,金属调节转录因子1(MTF1)的mRNA表达水平、硫辛酸合成酶(LIAS)的mRNA和蛋白表达水平增高,而血管内皮生长因子受体A(VEGFA)的mRNA表达水平、肿瘤蛋白p53(TP53)的mRNA和蛋白表达水平降低(P<0.05)。(3)与sh-WDHD1对照组及sh-WDHD1/DMSO组相比,sh-WDHD1/ATTM组HONE-1细胞和HK-1细胞的铜含量下降,HONE-1细胞在48 h的增殖活力增强,HK-1细胞在24 h、48 h的增殖活力增强(P<0.05)。结论敲低WDHD1可降低鼻咽癌细胞的增殖活性,增加细胞内铜含量蓄积,上调铜死亡促进因子LIAS、MTF1的表达,下调铜转运关键因子VEGFA及铜代谢关键因子TP53的表达。TP53和LIAS可能在WDHD1影响鼻咽癌细胞的铜蓄积和铜死亡进程中发挥重要作用。Objective To investigate the effect of WD repeat and HMG⁃box DNA binding protein 1(WDHD1)knockdown on proliferation,copper content,and copper death⁃related factors in nasopharyngeal carcinoma cells.Methods Nasopharyngeal carcinoma cells(HK⁃1 cell and HONE⁃1 cell)were assigned to control group,knockdown vector group(the sh⁃NC group),or WDHD1 knockdown group(the sh⁃WDHD1 group).The control group received no treatment,whereas negative control lentivirus and sh⁃WDHD1 RNA lentivirus were transfected in the sh⁃NC and sh⁃WDHD1 groups,respectively.After successful transfection,proliferation activity,copper content,and mRNA and protein expressions of copper death⁃related genes of HK⁃1 and HONE⁃1 cells were detected in various groups.HK⁃1 and HONE⁃1 cells of the sh⁃WDHD1 group were assigned to sh⁃WDHD1 control group,sh⁃WDHD1/DMSO group,or sh⁃WDHD1/ATTM group.Complete medium,complete medium containing DMSO,and complete medium containing copper chelating agent ATTM were employed to perform culture,respectively,and then copper content in cells was detected.Results(1)Compared with the control and sh⁃NC groups,proliferation activity of HONE⁃1 cell in the sh⁃WDHD1 group was decreased in 24 and 48 hours,and proliferation activity of HK⁃1 cell was decreased in 48 hours(P<0.05).(2)In HONE⁃1 and HK⁃1 cells,compared with the control and sh⁃NC groups,the sh⁃WDHD1 group yielded an increased copper content,an elevated mRNA expression of metal regulatory transcription factor 1(MTF1),and elevated mRNA and protein expressions of lipoic acid synthetase(LIAS),whereas a decreased mRNA expression of vascular endothelial growth factor A(VEGFA),and decreased mRNA and protein expressions of tumor protein p53(TP53,P<0.05).(3)Compared with the sh⁃WDHD1 control group and the sh⁃WDHD1/DMSO group,copper content of HONE⁃1 and HK⁃1 cells were decreased in the sh⁃WDHD1/ATTM group,whereas HONE⁃1 cell proliferation activity was increased in 48 hours,and HK⁃1 cell proliferation activity was i

关 键 词:鼻咽癌 铜死亡 敲低WD重复序列和HMG框DNA结合蛋白1 硫辛酸合成酶 肿瘤蛋白p53 

分 类 号:R34[医药卫生—基础医学]

 

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