下调基因PTTG1表达对前列腺癌LNCaP-AI细胞增殖、侵袭和凋亡的影响
发布时间：2018-03-20 11:49
本文选题：前列腺癌 切入点：垂体肿瘤转化基因 出处：《中华男科学杂志》2017年07期 论文类型：期刊论文
【摘要】：目的:探讨下调垂体肿瘤转化基因1(PTTG1)表达对雄激素非依赖性前列腺癌LNCa P-AI细胞增殖、侵袭、凋亡,以及对雄激素拮抗剂敏感性的影响。方法:LNCa P-AI细胞转染靶向PTTG1基因的siRNA,MTT法检测细胞增殖,Transwell法检测细胞侵袭,流式细胞术检测细胞凋亡,Western印迹检测PTTG1、p-Akt、p-ERK等蛋白表达水平,琼脂糖凝胶电泳法检测PTTG1 mRNA表达水平。结果:siRNA有效抑制了PTTG1的表达;下调PTTG1表达有效抑制了LNCa P-AI细胞在去雄激素培养液中的增殖,24、48、72 h抑制率分别为(19.47±2.12)%、(24.01±2.13)%、(48.02±2.22)%,组间比较有显著性差异(P0.05);降低其侵袭力,Transwell试验显示,对照组、转染24、48、72 h后穿过聚碳酸酯膜细胞个数分别为111.11±13.47、74.67±9.85、56.44±8.66、37.33±6.14,组间比较有显著性差异(P0.01);siRNA-PTTG1转染LNCa P-AI细胞后,空白对照组、转染24、48、72 h后凋亡率分别为(2.17±0.49)%、(18.32±0.94)%、(19.94±1.30)%、(21.73±1.88)%,各组间比较有显著性差异(P0.05)。siRNA联合氟他胺组对LNCa P-AI增殖的抑制作用和促凋亡作用显著强于siRNA或者氟他胺组,并呈氟他胺浓度相关性;50 nmol/L氟他胺、siRNA联合50 nmol/L氟他胺对LNCa P-AI的抑制率分别为(27.13±3.52)%、(67.51±5.13)%,两组间比较有显著性差异(P0.05);凋亡率分别为(3.94±0.48)%、(19.93±1.72)%,两组间比较有显著性差异(P0.05);100 nmol/L氟他胺、siRNA联合100 nmol/L氟他胺的抑制率分别为(43.72±3.90)%、(73.19±4.78)%,两组间比较有显著性差异(P0.05);凋亡率分别为(5.33±0.66)%、(23.43±1.76)%,两组间比较有显著性差异(P0.05)。结论:siRNA下调PTTG1表达能够抑制LNCa P-AI的增殖和侵袭,促进凋亡,提高了LNCa P-AI细胞对雄激素拮抗剂氟他胺的敏感性,抑制PTTG1表达可能会强化雄激素剥夺治疗晚期前列腺癌的作用。
[Abstract]:Objective: To investigate the downregulation of pituitary tumor transforming gene 1 (PTTG1) expression on the proliferation of androgen independent prostate cancer LNCa P-AI cells, invasion, apoptosis, and the effect of androgen antagonist sensitivity. Methods: siRNA LNCa cells were transfected with P-AI targeting PTTG1 gene, MTT cell proliferation assay, cell invasion was detected by Transwell assay. Flow cytometry, PTTG1 p-Akt, Western blot, the expression level of p-ERK protein, detected the expression of PTTG1 mRNA by agarose gel electrophoresis. Results: siRNA inhibited the expression of PTTG1 down regulated PTTG1 expression; effective suppression of LNCa in P-AI cells to androgen in the culture medium of proliferation, the inhibition rate of H 24,48,72 respectively (19.47 + 2.12)% and (24.01 + 2.13)% and (48.02 + 2.22)%, there were significant difference between two groups (P0.05); reduce the invasion force, Transwell test showed that the control group, h after transfection of 24,48,72 through poly carbon 閰搁叝鑶滅粏鑳炰釜鏁板垎鍒负111.11卤13.47,74.67卤9.85,56.44卤8.66,37.33卤6.14,缁勯棿姣旇緝鏈夋樉钁楁，
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