摘要
目的探究PTEN(phosphatase and tensin homologue deleted on chromosome ten)过表达对卡博替尼治疗肾透明细胞癌作用的影响。方法构建PTEN过表达细胞系以及抑制细胞内PTEN表达,将细胞分为NC、NC+C、PI、PI+C、PO和PO+C组,MTT方法检测卡博替尼对细胞增殖的影响。qPCR方法检测凋亡相关蛋白p53、BCL-2相关蛋白X(BCL-2 Associated X,BAX)、B细胞淋巴瘤基因2(B cell lymphoma gene 2,Bcl-2)在基因水平上表达量的改变,Western blot检测细胞内PI3K/AKT-mTOR信号通路的活性以及凋亡相关蛋白p53、BAX、Bcl-2表达量的改变,以及三羧酸循环关键酶ACO2和SUCLG2表达量的变化。结果卡博替尼能够明显抑制透明细胞癌的增殖(P<0.05),细胞内PI3K/AKT-mTOR信号通路的活性也受到明显的抑制(P<0.05)。细胞内促凋亡蛋白BAX的表达显著升高(P<0.05),而抗凋亡蛋白Bcl-2表达则显著降低(P<0.05),基因水平检测也呈现出类似的趋势,表明细胞内凋亡过程受到激活。此外,细胞内三羧酸循环关键酶乌头酸酶2(aconitase2,ACO2)表达显著增加(P<0.05),琥珀酰辅酶A合成酶(succinyl-CoA synthase2,SUCLG2)表达显著降低(P<0.05),表明三羧酸循环也受到抑制。PTEN过表达会增加卡博替尼抗肿瘤细胞的作用而表达抑制后则会减弱这种作用。结论卡博替尼通过抑制细胞内PI3K/AKT-mTOR信号通路,活化细胞凋亡途径,抑制细胞内三羧酸循环过程,从而发挥抗肿瘤作用,PTEN过表达会增强卡博替尼的抗肿瘤作用。
Objective To explore the effect of PTEN overexpression on the treatment of the renal clear cell carcinoma with cabozantinib. Methods Firstly PTEN was overexpressed or inhibited in ccRCC cells,and cells were divided into NC,NC+C,PI,PI+C,PO,PO+C groups.The effect of cabozantinib on proliferation of cells were observed by using MTT assay,the expression levels of apoptosis related genes p53,BAX,Bcl-2 were deected by using qPCR analysis.Activity of PI3K/AKT-mTOR signaling pathway,expression levels of apoptosis related proteins p53,BAX,Bcl-2 and expression levels of key enzymes in TCA cycle ACO2 and SUCLG2 were detected by using Western blotting analysis. Results Proliferation of ccRCC was significantly inhibited after cabozantinib treatment ( P <0.05),the activity of cellular PI3K/AKT-mTOR signaling pathway was also significantly inhibited ( P <0.05).The expression of BAX was significantly increased and the expression of Bcl-2 was significantly inhibited ( P <0.05) at protein level as well as at transcription level,activate cellular apoptosis process.Besides,the expression levels of key enzymes in TCA cycle were significantly changed.The expression of ACO2 was significantly increased ( P <0.05) and the expression of SUCLG2 was significantly decreased ( P <0.05).PTEN overexpression or inhibition would enlarge or reduce the effect of cabozantinib on ccRCC. Conclusion Cabozantinib performed an anti-tumor effect via inhibition of PI3K/AKT-mTOR signaling pathway,activation of cellular apoptosis process and inhibition of celluar TCA cycle.PTEN overexpression would enhance the anti-tumor effect of cabozantinib.
作者
肖志中
韩光
周娟
陈旭义
王振国
张景童
刘洋
Xiao Zhizhong;Han Guang;Zhou Juan;Chen Xuyi;Wang Zhenguo;Zhang Jingtong;Liu Yang(Department of Urinary Surgery,Specialty Medical Center of Logistics University of Chinese People's Armed Police Force,Tianjin 300162;Department of Brain Center,Specialty Medical Center of Logistics University of Chinese People's Armed Police Force,Tianjin 300162;Department of Health and Education,Specialty Medical Center of Logistics University of Chinese People's Armed Police Force,Tianjin 300162;Second Brigade,Logistics University of Chinese People's Armed Police Force,Tianjin 300162;Department of Neurology,Shanghai Fourth People's Hospital,Shanghai 200000)
出处
《遵义医学院学报》
2019年第1期57-63,共7页
Journal of Zunyi Medical University
基金
天津市科技计划资助项目(NO:16ZXHLSY00120)
关键词
透明细胞癌
PTEN
细胞凋亡
三羧酸循环
clear cell renal cell carcinoma
PTEN
apoptosis
tricarboxylic acid cycle
