癌变·畸变·突变 ›› 2015, Vol. 27 ›› Issue (3): 177-181,186.doi: 10.3969/j.issn.1004-616x.2015.03.004

• 论著 • 上一篇    下一篇

亲环素A过表达对高温、紫外线和顺铂引起的H1299细胞凋亡的影响

阎瑾, 赵晓婷, 江妹, 顾勐, 岳文涛   

  1. 首都医科大学附属北京胸科医院细胞生物实验室, 北京 通州 101100
  • 收稿日期:2015-02-02 修回日期:2015-04-07 出版日期:2015-05-30 发布日期:2015-05-30
  • 通讯作者: 岳文涛,E-mail:Yuewt2000@yahoo.coms E-mail:Yuewt2000@yahoo.coms
  • 作者简介:阎瑾,E-mail:cilovesunrise@163.com
  • 基金资助:

    北京市教育委员会科技发展计划(km201210025024);高等学校博士学科点专项科研基金(20121107120012)

High temperature, ultraviolet light and cisplatin caused apoptosis of H1299 cell line which over expressed cyclophilin A

YAN Jin, ZHAO Xiaoting, JIANG Mei, GU Meng, YUE Wentao   

  1. Department of Cellular & Molecular Biology, Beijing Chest Hospital, Capital Medical University, Tongzhou 101100, Beijing, China
  • Received:2015-02-02 Revised:2015-04-07 Online:2015-05-30 Published:2015-05-30

摘要:

目的:探讨人肺癌细胞株H1299过表达亲环素A(CyPA)后对高温、紫外线和顺铂作用引起的细胞凋亡的影响。方法:采用重组慢病毒介导构建CyPA过表达H1299细胞株H1299 CyPA OE,并用Western blot方法验证CyPA过表达细胞株是否成功构建。培养H1299和H1299 CyPA OE细胞株,分别进行高温(44 ℃处理60 min)、紫外线(90 μW/cm2处理60 min)和顺铂(37.5、75和150 μg/mL作用2 h)处理后,用高内涵分析平台检测细胞的晚期凋亡。结果:H1299 CyPA OE细胞能检测到绿色荧光,且其CyPA蛋白表达较H1299细胞明显增高,表明H1299 CyPA OE细胞株构建成功。在不同顺铂浓度下,H1299 CyPA OE细胞凋亡率较H1299细胞均明显降低(P均<0.05),而在高温和紫外线作用下,H1299 CyPA OE细胞凋亡率与H1299细胞相比差异均无统计学意义(P均>0.05)。结论:CyPA过表达对顺铂诱导的凋亡有抵抗作用,表明CyPA可作为潜在的联合化疗的基因靶点,有望为临床个体化治疗提供新的思路。

关键词: 亲环素A, 凋亡, 顺铂, H1299细胞, 肺癌

Abstract:

OBJECTIVE: This study aimed to explore the effects of cyclophilin A (CyPA) over-expression in human lung adenocarcinoma cell line H1299 after treatments of high temperature, ultraviolet (UV) and cisplatin-induced apoptosis. METHODS:We set up H1299 CyPA over-expression cell lines via lentiviral transfection and used fluorescence microscope and Western blot analysis to analyze the expression of the target gene. H1299 and H1299 CyPA OE cell lines were cultured and exposed to treatments of high temperature, UV and cisplatin. High intension analysis platform was used to detect apoptosis of the two cell lines. RESULTS:Green fluorescent could be detected in H1299 CyPA OE cell line, and the protein CyPA expression was much higher than H1299 cell line, indicating that H1299 CyPA OE cell line was set up successfully. The apoptosis rate of H1299 CyPA OE cell line was much lower than H1299 cell line under different concentrations of cisplatin(P<0.05), but meanwhile the difference of apoptosis rates of H1299 CyPA OE cell line and H1299 cell lines showed no statistical significance(P>0.05). CONCLUSION:The over-expression of CyPA could confer resistance to apoptosis induced by cisplatin. CyPA may act as a potential gene target and provide new strategies of combined chemotherapy for lung cancer.

Key words: cyclophilin A, apoptosis, cisplatin, H1299 cell, lung cancer

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