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目的 :观察过表达丝裂原诱导基因6(mitogen-inducible gene 6,MIG6)对子宫内膜腺癌细胞(Ishikawa cell)增殖、凋亡和侵袭能力的影响。方法:脂质体法介导MIG6过表达质粒转染Ishikawa细胞,采用实时荧光定量PCR和Western blot检测转染前、后MIG6表达变化;流式细胞仪检测细胞凋亡的变化;Western blot检测凋亡相关蛋白和周期蛋白的表达;5-溴脱氧尿嘧啶核苷(Brd U)标记法检测细胞增殖情况;Transwell检测细胞侵袭能力的变化。结果:转染MIG6过表达质粒后,Ishikawa细胞中MIG6 m RNA和蛋白的水平分别为对照组的24.2倍和2.8倍,差异具有统计学意义(P<0.05)。增加Ishikawa细胞中MIG6表达后,裂解的半胱氨酰天冬氨酸蛋白酶-3(cleaved caspase 3)表达显著上升(P<0.05);早期凋亡率、晚期凋亡率分别较对照组升高1.7倍(P<0.01)、2.7倍(P<0.05);Cyclin D1蛋白表达明显下降(P<0.05),ERK蛋白磷酸化水平下降,p ERK/ERK比值降低(P<0.05);细胞增殖率由(43.4±2.4)%下降至(30.5±4.3)%(P<0.05);穿过Transwell膜的细胞数从(36.2±6.9)个下降至(26.2±3.8)个(P<0.05)。结论 :MIG6表达上调能抑制Ishikawa细胞的增殖水平和侵袭能力,并促进其凋亡。
Objective: To observe the effect of overexpression of mitogen-inducible gene 6 (MIG6) on the proliferation, apoptosis and invasion of endometrial adenocarcinoma cells (Ishikawa cells). Methods: Lipofectamine was used to transfect MIG6 overexpression plasmid into Ishikawa cells. The changes of MIG6 expression before and after transfection were detected by real-time fluorescence quantitative PCR and Western blot. The changes of apoptosis were detected by flow cytometry. The expression of death related protein and cyclin was detected. The proliferation of cells was detected by BrdU labeling. The invasion ability of cells was detected by Transwell assay. Results: After MIG6 overexpression plasmid was transfected, the levels of MIG6 mRNA and protein in Ishikawa cells were 24.2 and 2.8 times higher than those in control group, respectively. The difference was statistically significant (P <0.05). After MIG6 expression was increased in Ishikawa cells, the cleaved caspase 3 expression was significantly increased (P <0.05). The early apoptosis rate and the late apoptosis rate were significantly higher than those in the control group 1.7 fold (P <0.01), 2.7 fold (P <0.05); the expression of Cyclin D1 was significantly decreased (P <0.05); the phosphorylation of ERK was decreased; the ratio of p ERK / ERK was decreased (43.4 ± 2.4)% to (30.5 ± 4.3)% (P <0.05). The number of cells passing through Transwell membrane decreased from (36.2 ± 6.9) to (26.2 ± 3.8) (P <0.05). Conclusion: Up-regulation of MIG6 can inhibit Ishikawa cells proliferation and invasion, and promote apoptosis.