过表达丝裂原诱导基因6对子宫内膜癌细胞凋亡和侵袭力的影响

发布时间：2018-09-07 21:12

【摘要】：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(P0.05)。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(P0.05);早期凋亡率、晚期凋亡率分别较对照组升高1.7倍(P0.01)、2.7倍(P0.05);Cyclin D1蛋白表达明显下降(P0.05),ERK蛋白磷酸化水平下降,p ERK/ERK比值降低(P0.05);细胞增殖率由(43.4±2.4)%下降至(30.5±4.3)%(P0.05);穿过Transwell膜的细胞数从(36.2±6.9)个下降至(26.2±3.8)个(P0.05)。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。
[Abstract]:Aim: to investigate the effects of overexpression of mitogen inducible gene 6 (mitogen-inducible gene 6) on the proliferation, apoptosis and invasion of endometrial adenocarcinoma cell line (Ishikawa cell). Methods: MIG6 overexpression plasmids were transfected into Ishikawa cells by liposome method. The changes of MIG6 expression before transfection and after transfection were detected by real-time fluorescence quantitative PCR and Western blot. The expression of apoptosis-related protein and cyclin were detected by flow cytometry and the expression of apoptosis-related protein and cyclin were detected by Western blot. The proliferation of cells was detected by (Brd U) labeling method and the change of cell invasion ability was detected by Transwell. Results: the levels of MIG6 m RNA and protein in Ishikawa cells were 24.2 times and 2.8 times as much as those in the control group after transfection of MIG6 overexpression plasmid. The difference was statistically significant (P0.05). After increasing the expression of MIG6 in Ishikawa cells, the expression of cysteine aspartate protease-3 (cleaved caspase 3) increased significantly (P0.05), and the rate of early apoptosis increased significantly (P < 0.05). Compared with the control group, the late apoptotic rate increased 1.7-fold (P0.01) 2.7-fold (P0.05) the expression of Cyclin D1 protein decreased significantly (P0.05), the ratio of cell proliferation decreased from (43.4 卤2.4)% to (30.5 卤4.3)% (P0.05), the number of cells passing through the Transwell membrane decreased from (36.2 卤6.9) to (26.2 卤3.8) (P0.05), the cell proliferation rate decreased from (43.4 卤2.4)% to (30.5 卤4.3)% (P0.05), and the cell proliferation rate decreased from (43.4 卤2.4)% to (30.5 卤4.3)% (P0.05). Conclusion the upregulation of the expression of mig 6 can inhibit the proliferation and invasion ability of Ishikawa cells and promote its apoptosis.
【作者单位】：南京医科大学第一附属医院妇科;苏州大学附属第一医院妇产科;
【基金】：国家自然科学基金(81070471) 江苏省“十二五”兴卫工程重点人才资助(RC201161)
【分类号】：R737.33

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