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作 者:陈坤 张洁清[1] 李力[1] Chen Kun;Zhang Jieqing;Li Li(Department of Gynecology and Oncology,Affiliated Tumor Hospital of Guangxi Medical University,Nanning 530000)
机构地区:[1]广西医科大学附属肿瘤医院妇瘤科,南宁530000
出 处:《现代妇产科进展》2021年第4期270-272,共3页Progress in Obstetrics and Gynecology
基 金:国家自然科学基金资助项目(No:81160318);广西自然科学基金资助项目(No:2013GXNSFAA019219)。
摘 要:目的:探讨沉默PI3K基因对雌激素刺激下的子宫内膜癌细胞增殖、迁移、凋亡的影响。方法:培养子宫内膜癌Ishikawa细胞,构建LV-PI3K-RNAi慢病毒载体转染Ishikawa细胞。实验分组Control组未经任何处理的Ishikawa细胞;Ishikawa组经E_(2)刺激的Ishikawa细胞;NC组经E_(2)刺激并转染阴性对照慢病毒的Ishikawa细胞;PI3Ki组经E_(2)刺激并转染LV-PI3K-RNAi慢病毒的Ishikawa细胞。Real-time PCR、Western blot法检测各组VEGF、bFGF、PI3K mRNA及蛋白表达水平,MTT法、流式细胞仪分别检测细胞增殖能力及凋亡的情况。结果:与Ishikawa组、NC组比较,PI3Ki组的VEGF、bFGF、PI3K mRNA及蛋白表达水平明显降低,细胞增殖能力显著降低,凋亡率增高,差异均有统计学意义(P<0.05)。结论:PI3K基因低表达可干扰E_(2)在基因、蛋白水平激活Ishikawa细胞产生VEGF、bFGF,从而下调E_(2)对子宫内膜癌细胞增殖和抑制凋亡的影响。Objective:To investigate the effect of PI3K gene silencing on proliferation and apoptosis of endometrial cancer cells.Methods:Ishikawa cells were cultured and transfected with LV-PI3K-RNAi lentivirus vector.The experiment was divided into four groups:Control group:Ishikawa cells without any treatment,Ishikawa group:Ishikawa cells stimulated by E_(2),NC group:Ishikawa cells stimulated by E_(2)and transfected with lentivirus negative control,PI3Ki group:Ishikawa cells stimulated by E_(2)and transfected with LV-PI3K-RNAi lentivirus.Real-time PCR and Western blot were used to detected the mRNA and protein expressions of VEGF,bFGF and PI3K.MTT assay was used to evaluate the activity of cell proliferation,and the cell apoptosis rate were detected by flow cytometry.Results:Compared with the other experimental groups,the ability of proliferation was significantly reduced in the PI3K silence group,and the apoptosis rate of this group was increased(P<0.05).Conclusion:Low expression of PI3K gene can interfere E_(2)in activating Ishikawa cells to produce VEGF and bFGF in gene and protein levels,thus downregulate the effect of E_(2)on proliferation and apoptosis of endometrial cancer cell.
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