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作 者:赵楠[1] 邹丽娟[2] 苏晗[3] 薛晖[1] ZHAO Nan;ZOU Lijuan;SU Han;XUE Hui(Department of Gynecology,The First Hospital of China Medical University,Shenyang 110001,China;Clinical Laboratory,The Fourth Affiliated Hospital of China Medical University,Shenyang 110032,China;Equipment Section,Shengjing Hospital of China Medical University,Shenyang 110004,China)
机构地区:[1]中国医科大学附属第一医院妇科,沈阳110001 [2]中国医科大学附属第四医院检验科,沈阳110032 [3]中国医科大学附属盛京医院设备科,沈阳110004
出 处:《中国医科大学学报》2022年第8期725-729,共5页Journal of China Medical University
基 金:辽宁省教育厅2019年度科学研究项目(QN2019011)。
摘 要:目的 探讨泛素结合酶E2T (UBE2T)基因在卵巢癌组织中的表达水平及其对卵巢癌细胞增殖的调控作用和机制。方法 通过基因表达谱交互分析(GEPIA)网站分析UBE2T基因在卵巢癌组织与正常组织中的差异表达。收集52例卵巢癌组织和癌旁组织样本,采用实时荧光定量PCR (qRT-PCR)检测UBE2T基因在卵巢癌组织和癌旁组织中的差异表达;采用MTT法检测细胞增殖的改变;采用Western blotting检测FA/BRCA通路FANCF和FANCD2蛋白的表达;采用彗星实验检测DNA损伤。结果 qRT-PCR结果显示,UBE2T基因在卵巢癌组织中的表达明显高于癌旁组织(P <0.01)。转染cDNA-UBE2T至SKOV3细胞后UBE2T蛋白高表达,与阴性转染组比较,细胞增殖水平明显增加(P <0.05)。Western blotting结果显示,UBE2T过表达后可增加SKOV3细胞中FA/BRCA通路FANCF和FANCD2蛋白的表达(P <0.05)。彗星实验发现,UBE2T过表达可以抑制顺铂引起的卵巢癌细胞DNA损伤。结论 UBE2T基因在卵巢癌组织中高表达,UBE2T通过FA/BRCA通路调控卵巢癌细胞的增殖,其有望成为卵巢癌新的治疗靶点。Objective To investigate the expression level of the ubiquitin binding enzyme E2T(UBE2T) gene in ovarian cancer,and its regulatory effect and mechanism on the proliferation of ovarian cancer cells. Methods The differential expression of the UBE2T gene in ovarian cancer and normal tissues was analyzed using the Gene Expression Profiling Interactive Analysis(GEPIA) website. The differential expression of UBE2T gene in ovarian cancer and adjacent tissues was detected by real-time quantitative PCR(qRT-PCR). Cell proliferation was detected by MTT assay. To explore the mechanism of action,the expression levels of FANCF and FANCD2 proteins in the FA/BRCA pathway were detected by Western blotting. Comet assay was performed to detect DNA damage. Results The results of q RT-PCR showed that the expression of UBE2T gene in ovarian cancer tissues was significantly higher than that in normal tissues(P < 0.01).Compared with the empty vector transfection group,the proliferation activity of the SKOV3 cells significantly increased after transfection of cDNA-UBE2T(P < 0.05). Overexpression of UBE2T subsequently increased the expression of FANCF and FANCD2 in the FA/BRCA pathway in SKOV3 cells(P < 0.05). Comet assay showed that UBE2T overexpression inhibited cisplatin-induced DNA damage in ovarian cancer. Conclusion UBE2T is highly expressed in ovarian cancer. The UBE2T gene regulates the proliferation of ovarian cancer cells through the FA/BRCA pathway and may be a potential new therapeutic target in ovarian cancer patients.
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