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RNA干扰YWHAZ基因对结肠癌HCT-8细胞增殖的影响 被引量:2
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作者 张成才 宋艳艳 +3 位作者 周梦娇 崔志倩 张西臣 邢沈阳 《吉林大学学报(医学版)》 CAS CSCD 北大核心 2015年第2期307-310,I0005,共5页
目的:探讨中RNA干扰YWHAZ基因对结肠癌HCT-8细胞增殖的影响,阐明其作用机制。方法:构建靶向基因YWHAZ的siRNA载体重组质粒pGPU6/GFP/Neo-YWHAZ-792、pGPU6/GFP/NeoYWHAZ-733、pGPU6/GFP/Neo-YWHAZ-612和pGPU6/GFP/Neo-YWHAZ-505作为RNA... 目的:探讨中RNA干扰YWHAZ基因对结肠癌HCT-8细胞增殖的影响,阐明其作用机制。方法:构建靶向基因YWHAZ的siRNA载体重组质粒pGPU6/GFP/Neo-YWHAZ-792、pGPU6/GFP/NeoYWHAZ-733、pGPU6/GFP/Neo-YWHAZ-612和pGPU6/GFP/Neo-YWHAZ-505作为RNA干扰质粒组,同时设立阴性对照质粒组(pGPU6/GFP/Neo-shNC质粒);采用荧光定量PCR检测YWHAZ mRNA的表达水平和表达抑制率;采用MTT法检测HCT-8细胞的增殖率。结果:倒置荧光显微镜下观察,细胞转染率为60%。荧光定量PCR检测,RNA干扰组YWHAZ mRNA表达水平低于阴性对照组(P<0.01),平均抑制率为52.86%。MTT检测,与阴性对照组比较,RNA干扰质粒组细胞增殖率明显降低(P<0.01),平均细胞增殖率为68.75%。结论:干扰YWHAZ可抑制该基因的转录,并抑制HCT-8细胞的增殖。 展开更多
关键词 ywhaz基因 结肠肿瘤 HCT-8细胞 RNA干扰
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YWHAZ gene contributes to the radioresistance of oral squamous cell carcinoma cells
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作者 Songling Hu Cong Chen +4 位作者 Hengheng Chen Xin Yu Xiaofei Li Yang Bai Chunlin Shao 《Radiation Medicine and Protection》 2024年第1期30-36,共7页
Objective:To investigate the contribution of YWHAZ gene on the radioresistance and metastasis ability of oral squamous cell carcinoma(OSCC)cells.Mathods:The relationship between the expression level of YWHAZ gene and ... Objective:To investigate the contribution of YWHAZ gene on the radioresistance and metastasis ability of oral squamous cell carcinoma(OSCC)cells.Mathods:The relationship between the expression level of YWHAZ gene and the survival of head and neck squamous cell carcinoma(HNSC)patients was analyzed using Gene Expression Profiling Interactive Analysis(GEPIA)database.A radioresistance cell line(CAL-27R)was constructed by irradiating CAL-27 cells with frac-tional doses.Cell survival was measured by colony formation assay.Cell migration and invasion were detected by transwell assay.The formation of yH2AX foci was detected by immunofluorescence assay,The protein expressions were detected by Westemn blot assay.In some experiments,CAL-27R cells were effectively transferred with siRNA YWHAZ(siYWHAZ).Results:GEPIA database showed that the expression level of YWHAZ in HNSC tumors was higher than that in adjacent normal tissues,and the HNSC patients with higher level of YWHAZ had a shorter survival.In vitro experiments demonstrated that the expression of YWHAZ protein in CAL-27 cells was lower than HSC-3 cells(t=18.89,P<0.01)and radioresistant CAL-27R cells(t=25.70,P<0.01).Knockdown of YWHAZ gene significantly increased radiation-induced cell killing effect,apoptosis induction,and yH2AX foci formation inCAL-27R and HSC-3 cells.Moreover,siRNA YWHAZ transfection also reduced the invasion and migration abilities of the irradiated CAL 27R[t=21.09,P<0.01(migration);t=6.16,P<0.05(invasion)]and HSC-3 cells[t=34.53,P<0.001(migration);t=4.92,P<0.05(invasion)]and attenuated radiation-induced expressions of metastasis-related proteins,Conclusion:YWHAZ contributes to the radioresistance of oral squamous cells and thus it may applicable to be a potential target for OSOC radiotherapy. 展开更多
关键词 Oral squamous cells Radiation sensitivity ywhaz gene Cell migration
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miR-451靶向调控肾小球系膜细胞Ywhaz基因的表达 被引量:6
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作者 查何 彭惠民 +5 位作者 马晶 周吉 王瑞敏 尹频 文利 张政 《中国生物制品学杂志》 CAS CSCD 2013年第8期1107-1112,共6页
目的探讨miR-451对肾小球系膜细胞Ywhaz基因表达的靶向调控。方法应用生物信息学技术对miR-451的靶基因进行预测,PCR法扩增靶基因3′UTR全长片段,插入荧光素酶报告载体pmiR-RB-REPORTTMvector中,构建荧光素酶报告质粒pmiR-Ywhaz-3′UTR... 目的探讨miR-451对肾小球系膜细胞Ywhaz基因表达的靶向调控。方法应用生物信息学技术对miR-451的靶基因进行预测,PCR法扩增靶基因3′UTR全长片段,插入荧光素酶报告载体pmiR-RB-REPORTTMvector中,构建荧光素酶报告质粒pmiR-Ywhaz-3′UTR。将miR-451 mimics和NC mimics分别转染高糖培养条件下的小鼠系膜细胞,将miR-451 inhibitor和NC inhibitor分别转染低糖培养条件下的小鼠系膜细胞,采用Real-time RT-PCR和Westernblot法检测miR-451和Ywhaz的表达水平;将miR-451 mimics和NC mimics分别与pmiR-Ywhaz-3′UTR共转染高糖培养条件下的小鼠系膜细胞,miR-451 inhibitor和NC inhibitor分别与pmiR-Ywhaz-3′UTR共转染低糖培养条件下的小鼠系膜细胞,检测各组细胞荧光素酶活性。结果 Ywhaz基因3′UTR区有11个碱基与miR-451种子序列匹配,其中7个碱基呈高度保守性;荧光素酶报告质粒经测序鉴定构建正确;miR-451在miR-451 mimics组呈高表达,在miR-451 inhibitor组表达降低;miR-451可抑制Ywhaz的蛋白表达;荧光素酶活性检测发现,miR-451可通过与Ywhaz3′UTR的结合,抑制Ywhaz的表达。结论 miR-451可直接靶向糖尿病相关基因Ywhaz,抑制其蛋白表达,为进一步研究miR-451调控早期糖尿病肾病发生发展的机制提供了依据。 展开更多
关键词 miR-451 ywhaz基因 靶基因 早期糖尿病肾病
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