PTEN通过AKT介导的细胞代谢途径抑制宫颈癌细胞的增殖  

PTEN inhibits proliferation of cervical cancer cells by AKT mediated cell metabolism

作　　者：陈昆仑[1,2] 刘莹[1,3] 李明利[1] 高艳娥[1] 高庆[1] 

机构地区：[1]西安交通大学医学院第二附属医院妇产科,陕西西安710004 [2]西安交通大学医学院第二附属医院干部病房,陕西西安710004 [3]第一附属医院妇产科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2013年第5期563-567,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81070536)~~

摘　　要：目的研究PTEN基因与宫颈癌细胞增殖能力的关系,以探索PTEN基因在宫颈癌发生发展中的可能作用及其机制。方法 Western blot检测PTEN基因在宫颈癌细胞系HeLa、SiHa、C33A和CasKi中的表达。利用脂质体法将人工过表达PTEN载体转染低表达PTEN的宫颈癌细胞系HeLa和C33A,通过MTT方法观察转染后PTEN过表达对宫颈癌细胞增殖的影响。采用多功能过程参数分析仪,检测转染PTEN基因后宫颈癌细胞培养基中乳酸、葡萄糖及谷氨酰胺的含量,观察PTEN基因过表达对宫颈癌细胞代谢的影响。Western blot检测PTEN过表达后宫颈癌细胞中丙酮酸激酶(PKM2)、6-磷酸果糖激酶2(PFKFB3)、谷氨酰胺酶(GLS)、AKT和磷酸化的AKT(pAKT)表达情况。结果与正常宫颈上皮细胞相比,PTEN基因在4种宫颈癌细胞系中低表达。与对照组相比,过表达PTEN的HeLa和C33A细胞克隆中,PTEN基因的水平明显升高。细胞计数和MTT法也显示,过表达PTEN能够明显抑制肿瘤细胞的增殖速率(P<0.05);过表达PTEN基因的细胞培养液中葡萄糖及谷氨酰胺含量显著增多,但乳酸含量减少;同时还发现PTEN抑制了AKT的磷酸化水平并使PKM2、PFKFB3及GLS的表达水平降低。结论 PTEN基因可通过AKT途径调节细胞代谢,从而调节宫颈癌细胞的增殖。PTEN基因有可能成为宫颈癌的诊断和治疗的新靶点。Objective To explore the relationship of phosphatase and tensin homolog （PTEN） with cell proliferation in cervical cancer ceils so as to further analyze the role of PTEN and its underlying mechanisms in the pathogenesis and progression of cervical cancer cells. Methods The expression of PTEN was detected in different cervical cancer cell lines by Western blotting. Then the overexpression vector of PTEN was constructed and transfected into cervical cancer cells with Lipofectemine 2000. The proliferative index was estimated by cell counting and MTT. The contents of glucose, glutamine and lactic acid were detected by the BioProfile FLEX analyzer to evaluate the effect of PTEN on the cell metabolism. The proteins related to cell metabolism were measured by Western blotting. Results The expression of PTEN was down-regulated in cervical cancer cell lines compared to that of normal cervical epithelium cells. The overexpression PTEN vector was transfected into HeLa and C33A cells, resulting in the up-regulation of PTEN. Enforced expression of PTEN inhibited the proliferation of cervical cancer cells, which was confirmed by cell counting and MTT （P〈0.05）. We also found that glucose and glutamine contents in the medium with PTEN-overexpressing cells were increased, but lactic acid content was decreased. Moreover, PTEN suppressed the expression of pAKT, PFKFB3, PKM2 and GLS in these cells. Conclusion PTEN regulates the proliferation of cervical cancer cells by AKT mediated cell metabolism, providing new insights into novel molecular target for the diagnosis and therapy of cervical cancer.

关 键 词：PTEN 宫颈癌 细胞代谢 AKT途径 丙酮酸激酶 6磷酸果糖激酶2 谷氨酰胺酶 

分 类 号：R33[医药卫生—人体生理学]