沉默PI3K基因对E2刺激后子宫内膜癌细胞增殖和凋亡的影响

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E₂刺激的Ishikawa细胞;NC组经E₂刺激并转染阴性对照慢病毒的Ishikawa细胞;PI3Ki组经E₂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E₂在基因、蛋白水平激活Ishikawa细胞产生VEGF、bFGF,从而下调E₂对子宫内膜癌细胞增殖和抑制凋亡的影响。

Effect of silencing PI3K gene on proliferation and apoptosis of endometrial cancer cells stimulated by E2

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 E₂,NC group:Ishikawa cells stimulated by E₂and transfected with lentivirus negative control,PI3Ki group:Ishikawa cells stimulated by E₂and 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 E₂in activating Ishikawa cells to produce VEGF and bFGF in gene and protein levels,thus downregulate the effect of E₂on proliferation and apoptosis of endometrial cancer cell.