FASN对Erlotinib耐药细胞株生长的影响及其机制

中国临床药理学与治疗学 ›› 2014, Vol. 19 ›› Issue (10): 1120-1125.

FASN对Erlotinib耐药细胞株生长的影响及其机制

谌茜, 李敏, 马倬, 吴丹丹, 倪萍, 翟溯澜, 李萍, 邹美娟, 王雪融

南京医科大学基础医学院药理学系,南京 210029,江苏

收稿日期:2013-03-22 修回日期:2014-06-10 出版日期:2014-10-26 发布日期:2014-10-29
通讯作者: 王雪融,女,博士,教授,研究方向:肿瘤药理。Tel: 13813385723 E-mail: wangxrwn@hotmail.com
作者简介:谌茜,女,硕士研究生,研究方向:肿瘤药理。 Tel: 15850559710 E-mail: chenxikaoyan@126.com
基金资助:
国家自然科学基金( 30873099; 81102458; 811172004)

Effects of FASN on the growth of erlotinib-resistant cell line and its mechanism

CHEN Xi, LI Min, MA Zhuo, WU Dan-dan, NI Ping, ZHAI Su-lan, LI Ping, ZOU Mei-juan, WANG Xue-rong

Department of Pharmacology, Nanjing Medical University, Nanjing 210029, Jiangsu, China

Received:2013-03-22 Revised:2014-06-10 Online:2014-10-26 Published:2014-10-29

摘要/Abstract

摘要： 目的探讨脂肪酸合酶(fatty acid synthase,FASN)在非小细胞肺癌(non-small-cell carcinoma,NSCLC)中,对厄洛替尼(Erlotinib)耐药细胞株生长的影响和其可能的机制。方法应用实时定量PCR技术以及Western blot技术分别检测HCC827-EP细胞株与HCC827-ER细胞株中的FASN mRNA和蛋白水平变化。利用小干扰RNA技术阻抑FASN表达后,以SRB法检测干扰前后对HCC827-EP/ER细胞株生长作用的影响。以及阻抑FASN表达后,用实时定量PCR技术和Western blot检测叉头蛋白O1(forkhead box protein O1 ,FoxO1)水平的变化。用活性形式的FoxO1腺病毒感染细胞后,以SRB法检测细胞的存活率。结果 Erlotinib耐药细胞株的FASN表达水平高于亲代细胞株;阻抑FASN的表达,抑制了耐药细胞株的生长;下调FASN的表达,FoxO1 mRNA及蛋白水平均有所升高。上调FoxO1的表达,抑制了耐药细胞株的生长。结论 FASN的高表达发生于Erlotinib耐药细胞株中,下调FASN表达可以抑制耐药细胞株的生长,其可能机制是抑制FASN表达上调了FoxO1,从而抑制了细胞的生长。

关键词: 非小细胞肺癌, 厄洛替尼, 脂肪酸合酶, 叉头蛋白O1

Abstract: AIM: To observe the effects of FASN on the proliferation and mechanism in erlotinib-resistant NSCLC cells. METHODS: Dected protein and mRNA expression levels of FASN by Western blot and quantitative RT-PCR in parental HCC827 cells and paired erlotinib-resistant HCC827-ER cells. The 5-days growth curve of HCC827-EP/ER were determined by SRB assay,after downregulating of FASN with small-interference RNA,Meanwhile, dected protein and mRNA levels of FoxO1 .Cell numbers in 5 days were measured by SRB assay after HCC827-ER transfecting with active form of FoxO1 adenovirus. RESULTS: The expression of FASN in HCC827-ER cells increased compared with paired HCC827-EP cells. And downregulation of FASN expression inhibited the growth of HCC827-ER cells (P<0.05). Protein and mRNA expression levels of FoxO1 increased after downregulating FASN.Upregulation of FoxO1 inhibited the growth of HCC827-ER cells.CONCLUSION: FASN expressed in a higher lever in HCC827-ER cells compared with HCC827-EP cells.Downregulation of FASN expression inhibited the growth of HCC827-ER cells,possibly through increased FoxO1 expression.

Key words: NSCLC, Erlotinib, FASN, FoxO1

中图分类号:

R965.2

谌茜, 李敏, 马倬, 吴丹丹, 倪萍, 翟溯澜, 李萍, 邹美娟, 王雪融. FASN对Erlotinib耐药细胞株生长的影响及其机制[J]. 中国临床药理学与治疗学, 2014, 19(10): 1120-1125.

CHEN Xi, LI Min, MA Zhuo, WU Dan-dan, NI Ping, ZHAI Su-lan, LI Ping, ZOU Mei-juan, WANG Xue-rong. Effects of FASN on the growth of erlotinib-resistant cell line and its mechanism[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(10): 1120-1125.

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