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目的 :探讨垂体瘤转化基因(pituitary tumor transforming gene,PTTG)在乳腺癌细胞侵袭中的作用。方法 :应用蛋白质印迹法检测PTTG蛋白在不同侵袭性乳腺癌细胞中的表达。将靶向PTTG的si RNA片段转染至高侵袭性乳腺癌MDA231细胞(命名为si PTTG/MDA231)后,应用蛋白质印迹法检测细胞中PTTG蛋白的表达水平;应用Transwell法检测si PTTG/MDA231细胞的侵袭能力;表皮细胞生长因子(epithelial growth factor,EGF)刺激si PTTG/MDA231细胞后,应用蛋白质印迹法检测细胞中磷酸化-Akt(phospho-Akt,p-Akt)和磷酸化-AMP相关蛋白激酶相关的蛋白激酶5[phospho-AMP-related protein kinase(AMPK)-related protein kinase 5,p-ARK5]的表达水平。结果 :高侵袭性乳腺癌MDA231细胞中PTTG蛋白的表达水平高于低侵袭性乳腺癌MCF7和T47-D细胞(P值均<0.05)。si PTTG/MDA231细胞中PTTG蛋白的表达水平低于空白对照组(MDA231细胞未进行任何转染)和阴性对照组(MDA231细胞转染阴性对照片段scrambling,命名为scr/MDA231细胞)(P值均<0.05)。si PTTG/MDA231细胞的侵袭能力低于空白对照组和阴性对照组(P值均<0.01)。EGF刺激0、5和10 min后,scr/MDA231细胞中p-Akt和p-ARK5蛋白的表达水平逐渐上调(P值均<0.05),而si PTTG/MDA231细胞中p-Akt和p-ARK5蛋白的表达水平无显著改变(P值均>0.05)。结论 :高侵袭性乳腺癌细胞中PTTG蛋白的表达水平较高。抑制PTTG的表达后,MDA231细胞的侵袭能力下降,这一作用可能与p-Akt和p-ARK5蛋白的表达调控有关。
Objective: To investigate the role of pituitary tumor transforming gene (PTTG) in invasion of breast cancer cells. Methods: The expression of PTTG protein in different invasive breast cancer cells was detected by Western blotting. The siRNA targeting PTTG was transfected into the highly invasive breast cancer MDA231 cells (named as si PTTG / MDA231). The PTTG protein expression was detected by Western blotting. Transwell assay was used to detect the expression of si PTTG / MDA231 cells (PTTG / MDA231) cells were stimulated by epithelial growth factor (EGF). Western blotting was used to detect the expression of phospho-Akt (p-Akt) and phospho- Related protein kinase 5 (phospho-AMP-related protein kinase (AMPK) -related protein kinase 5, p-ARK5]. Results: The expression level of PTTG protein in highly aggressive breast cancer MDA231 cells was significantly higher than that in low invasive breast cancer MCF7 and T47-D cells (all P <0.05). si PTTG / MDA231cells PTTG protein expression levels were lower than the blank control group (MDA231 cells without any transfection) and negative control group (MDA231 cells transfected with a negative control fragment scrambling, named scr / MDA231 cells) (P value <0.05). The invasive ability of si PTTG / MDA231 cells was lower than that of blank control group and negative control group (all P <0.01). The expression of p-Akt and p-ARK5 in scr / MDA231 cells gradually increased after EGF was stimulated for 0, 5 and 10 min (P <0.05), while the expression of p-Akt and p-ARK5 in si PTTG / MDA231 cells There was no significant change in protein expression (P> 0.05). Conclusion: PTTG protein is highly expressed in highly aggressive breast cancer cells. Inhibition of PTTG expression, MDA231 cell invasion decreased, this effect may be related to p-Akt and p-ARK5 protein expression regulation.