PI3K/Akt抑制剂对鸡朊蛋白过表达DF-1细胞增殖、侵袭和凋亡的影响

doi:10.11843/j.issn.0366-6964.2015.10.018

畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (10): 1844-1850.doi: 10.11843/j.issn.0366-6964.2015.10.018

PI3K/Akt抑制剂对鸡朊蛋白过表达DF-1细胞增殖、侵袭和凋亡的影响

万学瑞，朱曼玲，杨润霞，刘桂林，刘磊，吴润^*

（甘肃农业大学动物医学院，兰州 730070）

收稿日期:2015-02-05 出版日期:2015-10-23 发布日期:2015-10-23
通讯作者: 吴润，男，教授，博士生导师，E-mail：wurun@gsau.edu.cn
作者简介:万学瑞（1979-），女，甘肃白银人，讲师，博士，主要从事预防兽医学研究工作，E-mail：383921499@qq.com
基金资助:
国家自然科学基金（31160510）；甘肃省自然科学研究基金计划项目（1107RJZA198）；甘肃农业大学动物医学院教研产学支持计划（JYCX-KX009）

Effects of PI3K/Akt Inhibitor on Proliferation，Invasion and Apoptosis of DF-1 Cells with Over-expression of Chicken PrP^C

WAN Xue-rui，ZHU Man-ling，YANG Run-xia，LIU Gui-lin，LIU Lei，WU Run^*

（College of Veterinary Medicine，Gansu Agricultural University，Lanzhou 730070，China）

Received:2015-02-05 Online:2015-10-23 Published:2015-10-23

摘要/Abstract

摘要：

为探讨PI3K/Akt信号通路在鸡细胞型朊蛋白（ChPrP^C）过表达的DF-1细胞（DF-1-PrP）增殖、黏附、侵袭和凋亡过程中的作用及其与ChPrP^C表达量的关系，以DF-1-PrP细胞为模型，空载体转染的DF-1-NC细胞和DF-1细胞为对照，分别用0、10、20、50、100、200 nmol•L^-1渥曼青霉素处理以抑制PI3K/Akt信号通路，检测细胞对鼠尾胶原的黏附能力，Transwell小室法检测细胞侵袭力，MTT法检测细胞增殖，流式细胞仪检测细胞凋亡，RT-PCR法检测PRNP基因转录量。结果显示，随着渥曼青霉素浓度的增加，DF-1-PrP、DF-1-NC和DF-1细胞的PRNP基因转录量均减少，其增殖、黏附、侵袭能力相应下降，而总凋亡率均升高；但在低于100 nmol•L^-1的同一渥曼青霉素浓度下，DF-1-PrP细胞增殖、黏附、侵袭能力始终高于对照组，而总凋亡率均低于对照组。本研究表明ChPrP^C的过量表达可促进DF-1细胞增殖、黏附和侵袭，抑制其凋亡，在以上过程中，PI3K/Akt信号通路可能具有重要的作用，渥曼青霉素能有效阻断这一通路，但PI3K/Akt信号通路并不是ChPrP^C调节细胞凋亡的唯一途径。

Abstract:

DF-1 cells with over-expression of chicken PrP^C (DF-1-PrP)，DF-1 cells with pcDNA3.1(DF-1-NC) and DF-1 cells were used as cell models in order to study the effect of PI3K/Akt pathway on DF-1-PrP cells proliferation，adhesion，invasion and apoptosis and its relationship with ChPrP^Cexpression.After cells were treated with 0，10，20，50，100 and 200 nmol•L^-1 Wortmannin，adhesion assay，transwell assay，MTT assay，flow cytometric assay and RT-PCR analyses were used to detect cell adhesion，invasion，proliferation，apoptosis and expression of PRNP mRNA，respectively.The data showed that the expression of PRNP mRNA of DF-1-PrP，DF-1-NC and DF-1 cells were all decreased with the increasing of wortmannin concentration，and adhesion，invasion and proliferation ability were reduced，but apoptosis rate were increased.Furthermore，under the same concentration of less than 100 nmol•L^-1，adhesion，invasion and proliferation ability of DF-1-PrP cells was higher than DF-1-NC and DF-1 cells，apoptosis rate was lower.All these results indicated that over-expression of ChPrP^C promoted DF-1-PrP cells adhesion，invasion and proliferation and inhibited apoptosis，in which processes PI3K/Akt pathway may play an important role.PI3K/Akt pathway can be effectively blocked by wortmannin，but it may not be the only pathway that ChPrP^C regulated cell apoptosis.

中图分类号:

万学瑞，朱曼玲，杨润霞，刘桂林，刘磊，吴润. PI3K/Akt抑制剂对鸡朊蛋白过表达DF-1细胞增殖、侵袭和凋亡的影响[J]. 畜牧兽医学报, 2015, 46(10): 1844-1850.

WAN Xue-rui，ZHU Man-ling，YANG Run-xia，LIU Gui-lin，LIU Lei，WU Run. Effects of PI3K/Akt Inhibitor on Proliferation，Invasion and Apoptosis of DF-1 Cells with Over-expression of Chicken PrP^C[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(10): 1844-1850.

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参考文献

［1］OSAKI M，OSHIMURA M，ITO H.PI3K-Akt pathway：its functions and alterations in human cancer［J］.Apoptosis，2004，9(6)：667-676.
［2］PRUSINER S B.Novel proteinaceous infectious particles cause scrapie［J］.Science，1982，216(4542)：136-144.
［3］JI H F，ZHANG H Y.A comparative molecular dynamics study on thermostability of human and chicken prion proteins［J］. Biochem Biophys Res Commun，2007，359(3)：790-794.
［4］BROWN D R，CLIVE C，HASWELL S J.Antioxidant activity related to copper binding of native prion protein［J］. J Neurochem，2001，76(1)：69-76.
［5］RACHIDI W，MANGÉ A，SENATOR A，et al.Prion infection impairs copper binding of cultured cells［J］. J Biol Chem，2003，278(17)：14595-14598.
［6］PAN Y，ZHAO L，LIANG J，et al.Cellular prion protein promotes invasion and metastasis of gastric cancer［J］.FASEB J，2006，20(11)：1886-1888.
［7］MEHRPOUR M，CODOGNO P.Prion protein：From physiology to cancer biology［J］.Cancer Lett，2010，290(1)：1-23.
［8］DIARRA-MEHRPOUR M，ARRABAL S，JALIL A，et al.Prion protein prevents human breast carcinoma cell line from tumor necrosis factor alpha-induced cell death［J］.Cancer Res，2004，64(2)：719-727.
［9］ROUCOU X，GIANNOPOULOS P N，ZHANG Y，et al.Cellular prion protein inhibits proapoptotic Bax conformational change in human neurons and in breast carcinoma MCF-7 cells［J］.Cell Death Differ，2005，12(7)：783-795.
［10］KIM B H，LEE H G，CHOI J K，et al.The cellular prion protein(PrP^C) prevents apoptotic neuronal cell death and mitochondrial dysfunction induced by serum deprivation［J］.Brain Res Mol Brain Res，2004，124(1)：40-50.
［11］DIAO X L，WU R，LIU L，et al.Expression patterns of PrP gene during chicken embryo development［J］.Asian J Anim Vet Adv，2011，7：199-204.
［12］刁小龙.鸡朊蛋白参与 DF-1 细胞增殖粘附的功能研究［D］.兰州：甘肃农业大学，2012：60.
DIAO X L.The function research of ChPrP^C involved in the processes of DF-1 cells proliferation and adhesion［D］.Lanzhou：Gansu Agricultural University，2012：60.(in Chinese)
［13］朱曼玲，吴润，于宏伟，等.鸡Akt基因mRNA SYBR Green Ⅰ实时荧光定量RT-PCR检测方法的建立［J］.中国兽医科学，2014，44(7)：758-764.
ZHU M L，WU R，YU H W，et al.Establishment of SYBR Green Ⅰ real-time RT-PCR for detection of chicken Akt gene mRNA［J］.Chinese Veterinary Science,2014，44(7)：758-764.(in Chinese)
［14］王晓南，吴青，张连生，等.PI3K/Akt抑制剂渥曼青霉素对白血病细胞增殖和凋亡的影响［J］.中国肿瘤生物治疗杂志，2010，17(1)：82-87.
WANG X N，WU Q，ZHANG L S，et al.Effect of PI3K/Akt inhibitor wortmannin on proliferation and apoptosis of leukemia K562 cells［J］.Chinese Journal of Cancer Biotherapy，2010，17(1)：82-87.(in Chinese)
［15］LEE S，CHOI E J，JIN C，et al.Activation of PI3K/Akt pathway by PTEN reduction and PIK3CA mRNA amplification contributes to cisplatin resistance in an ovarian cancer cell line［J］.Gynecol Oncol，2005，97(1)：26-34.
［16］WEISE J，SANDAU R，SCHWARTING S，et al.Deletion of cellular prion protein results in reduced Akt activation，enhanced postischemic caspase-3 activation，and exacerbation of ischemic brain injury［J］.Stroke，2006，37(5)：1296-1300.
［17］葛伏林，吕焱，吴本俨，等.朊蛋白PrP^C及磷酸化Akt在胃癌中共表达及其临床意义［J］.军医进修学院学报，2011，32(9)：962-964.
GE F L，LV Y，WU B Y，et al.Co-expression of PrP^C and p-Akt in gastric cancer and its clinical significance［J］.Journal of Chinese PLA Postgarduate Medical School，2011，32(9)：962-964.(in Chinese)
［18］OHTA T，OHMICHI M，HAYASAKA T，et al.Inhibition of phosphatidylinositol 3-kinase increases efficacy of cisplatin in in vivo ovarian cancer models［J］.Endocrinology，2006，147(4)：1761-1769.
［19］POH T W，PERVAIZ S.LY294002 and LY303511 sensitize tumor cells to drug-induced apoptosis via intracellular hydrogen peroxide production independent of the phosphoinositide 3-kinase-Akt pathway［J］.Cancer Res，2005，65(14)：6264-6274.

PI3K/Akt抑制剂对鸡朊蛋白过表达DF-1细胞增殖、侵袭和凋亡的影响

Effects of PI3K/Akt Inhibitor on Proliferation，Invasion and Apoptosis of DF-1 Cells with Over-expression of Chicken PrP^C

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PI3K/Akt抑制剂对鸡朊蛋白过表达DF-1细胞增殖、侵袭和凋亡的影响

Effects of PI3K/Akt Inhibitor on Proliferation，Invasion and Apoptosis of DF-1 Cells with Over-expression of Chicken PrPC

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Effects of PI3K/Akt Inhibitor on Proliferation，Invasion and Apoptosis of DF-1 Cells with Over-expression of Chicken PrP^C