国际肿瘤学杂志

国际肿瘤学杂志 ›› 2015, Vol. 42 ›› Issue (6): 401-406.doi: 10.3760/cma.j.issn.1673-422X.2015.06.001

• 论著 •    下一篇

蒿甲醚激活PPARγ途径抑制小鼠Lewis肺癌细胞体外增殖和侵袭机制研究

付凤莲, 蒋永新, 程荫, 刘珊, 王红   

  1. 50118昆明医科大学第三附属医院干部医疗科（付凤莲），中西医结合肿瘤临床研究中心（蒋永新、刘珊、王红）；昆明医科大学海源学院病理教研室（程荫）
  • 收稿日期:2014-11-25 出版日期:2015-06-08 发布日期:2015-05-31
  • 通讯作者: 蒋永新，Email：598104374@qq.com E-mail:蒋永新，Email：598104374@qq.com
  • 基金资助:

    昆明医科大学2013年研究生创新基金(2013N15)

Artemether inhibits proliferation and invasion via the mediation of peroxisome proliferatoractivated receptorgamma activation pathway in Lewis lung cancer cells

Fenglian, Jiang Yongxin, Cheng Yin, Liu Shan, Wang Hong   

  1. Department of Special Medical Treatment, Third Affiliated Hospital of Kunming Medical Univeristy, Kunming 650118, China
  • Received:2014-11-25 Online:2015-06-08 Published:2015-05-31
  • Contact: Jiang Yongxin E-mail:蒋永新，Email：598104374@qq.com

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摘要： 目的研究青蒿素类衍生物蒿甲醚（ARE)对小鼠Lewis肺癌细胞株（LLC）体外生长和侵袭的影响，探讨其可能的作用机制。方法采用MTT法观察ARE对LLC细胞生长的抑制效应。Transwell侵袭实验检测对照组、ARE组、GW9662［过氧化物酶体增殖物激活受体γ（PPARγ）特异性抑制剂］组和GW9662+ARE组4组LLC细胞的侵袭力。实时PCR和Western blotting法检测4组细胞PPARγ、NFκB p65、Caspase3 mRNA和蛋白表达。结果ARE呈时间和剂量依赖性方式抑制LLC细胞生长，24、48、72 h ARE对LLC的IC50值分别为271.29、189.08、65.99 μg/ml。4组中ARE组的荧光值最低，为1 744.67，对照组为6 887.00，GW9662+ARE组为4 597.00，GW9662组为10 012.67，表明ARE组侵袭力最弱，而且相比对照组侵袭力明显减弱（t=12.411，P=0.000）。ARE组、GW9662组PPARγ mRNA相对表达量分别为2.276±0.534和0.362±0.206，与对照组相比差异均有统计学意义（t=4.785，P=0.001;t=2.395，P=0.044）；ARE组PPARγ蛋白表达量为27 688.33±3 593.06，比对照组（17 716.33±2 273.95）、GW9662+ARE组（21 816.00±1 644.07）高（t=5.159，P=0.001;t=3.038，P=0.016）。NFκB p65 mRNA表达量在GW9662+ARE组（0.346±0.149）最低，其次为ARE组（0.392±0.187），GW9662组最高（1.720±0.338），ARE组与对照组和GW9662组相比差异有统计学意义（t=3.592，P=0.007;t=7.851，P=0.000）。Caspase3 mRNA和蛋白在各组的表达差异没有统计学意义（F=1.181，P=0.376；F=0.647，P＞0.05）。结论蒿甲醚可以通过上调PPARγ表达抑制NFκB的途径来抑制LLC细胞体外增殖和侵袭，提示PPARγ可能为肺癌治疗提供一个新的靶点。

关键词: 青蒿素类, 肺肿瘤, 过氧化物酶体增殖物激活受体, 核因子κB受体活化因子

Abstract: ObjectiveTo investigate the inhibitory effect of artemether (ARE), one of artemisinin derivatives, on the growth and invasion of Lewis lung cancer cells (LCCs) in vitro and explore the possible mechanism. MethodsEffects of ARE on proliferation of LLCs were evaluated by MTT. The invasiveness was detected by Transwell invasion assay, including control, ARE, GW9662 ［the specific inhibitor of peroxisome proliferatoractivated receptor γ (PPARγ)］ and GW9662+ARE group. The expression levels of PPARγ, NFκB p65, Caspase3 mRNA and protein in abovementioned four groups were analyzed by RTPCR and Western blotting, respectively. ResultsARE could inhibit the proliferation of LLCs in time and dosedependent manner, the IC50 value of which in 24, 48, and 72 h were 271.29, 189.08, 65.99 μg/ml, respectively. The fluorescent value of ARE group was 1 744.67, and that of control group was 6 887.00, 4 597.00 of GW+ARE group, as well as 10 012.67 of GW9662 group. It manifested the invasiveness of ARE group was the weakest, which declined significantly compared with control group (t=12.411，P=0.000). The relative quantitative expressions of PPARγ mRNA in ARE and GW9662 group were 2.276±0.534 and 0.362±0.026, respectively. Compared with control group, PPARγ mRNA level in both of ARE and GW9662 group reached statistical significance (t=4.785, P=0.001; t=2.395, P=0.044). PPARγ protein expression in ARE group，GW9662+ARE group and control group were 27 688.33±3 593.06, 21 816.00±1 644.07, 17 716.33±2 273.95, respectively, which was higher in ARE group than that in control and GW+ARE group (t=5.159, P=0.001; t=3.038, P=0.016). NFκB p65 mRNA expression in GW9662+ARE group was 0.346±0.149, which in ARE group and GW9662 group were 0.392±0.187 and 1.720±0.338, respectively. The differences of NFκB p65 mRNA expression level between ARE, and control or GW9662 group were statistically significant (t=3.592, P=0.007; t=7.851, P=0.000). While, the differences of Caspase3 mRNA and protein expression levels among the four groups were not statistically significant (F=1.181, P=0.376; F=0.647, P＞0.05). ConclusionARE may restrain NFκB through upregulating PPARγ to inhibit the proliferation and invasive potential of LLC in vitro, which suggests that PPARγ may be a novel therapeutic target for lung cancer.

Key words: Artemisinins, Lung neoplasms, Peroxisome proliferators activated receptors, Receptor activator of nuclear factor-kappa B