国际肿瘤学杂志

国际肿瘤学杂志 ›› 2016, Vol. 43 ›› Issue (3): 161-166.doi: 10.3760/cma.j.issn.1673-422X.2016.03.001

• 论著 •    下一篇

S100A6基因干扰对A549肺腺癌细胞生物学行为的影响

南岩东, 姜华, 金发光, 杨拴盈   

  1. 710038 西安，第四军医大学唐都医院呼吸与危重症医学科（南岩东、姜华、金发光）；西安交通大学第二附属医院呼吸内科（杨拴盈）
  • 出版日期:2016-03-08 发布日期:2016-02-03
  • 通讯作者: 南岩东，Email： nanyandong2008@163.com E-mail:nanyandong2008@163.com
  • 基金资助:

    国家自然科学基金（81001040）

Influence of S100A6 gene RNA interference on the biological behaviors of A549 lung adenocarcinoma cells

Nan Yandong,Jiang Hua,Jin Faguang,,Yang Shuanying   

  1. Department of Respiratory and Critical Care Medicine, Tangdu Hospital, Fourth Military Medical University, Xi′an 710038, China
  • Online:2016-03-08 Published:2016-02-03
  • Contact: Nan Yandong, Email: nanyandong2008@163.com E-mail:nanyandong2008@163.com
  • Supported by:

    National Natural Science Foundation of China (81001040)

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1416

被引次数

5

摘要： 目的 探讨S100A6基因干扰对A549肺腺癌细胞生物学行为的影响。方法 构建S100A6基因RNAi载体，慢病毒转染A549肺腺癌细胞，共分为3组，①空载体对照组：转染不携带S100A6基因RNAi的空载质粒；②阴性对照组：未转染任何质粒；③S100A6 RNA干扰组：携带S100A6基因RNAi的质粒。应用实时聚合酶链反应（RTPCR）和Western blotting鉴定S100A6基因和蛋白表达；采用四甲基偶氮唑蓝、迁移试验和流式细胞仪分别检测细胞增殖、浸润、细胞周期及细胞凋亡等生物学特性。结果 S100A6基因干扰后A549肺腺癌细胞S100A6 mRNA的表达（0.009±0.001）较阴性对照组（0.049±0.005）和空载体对照组（0.030±0.006）均有明显下降，差异均有统计学意义（t=57.56，P=0.000；t=48.21，P=0.000）。S100A6基因干扰后A549肺腺癌细胞S100A6蛋白的表达（0.107±0.002）较阴性对照组（0.341±0.005）和空载体对照组（0.311±0.006）均有明显下降，差异均有统计学意义（t=37.34，P=0.000；t=27.51，P=0.001）。48 h细胞增殖能力：RNA干扰组（0.230±0.008）较阴性对照组（0.292±0.038）和空载体对照组（0.307±0.013）降低，差异均有统计学意义（t=25.31，P=0.003；t=29.42，P=0.001）。细胞浸润能力：RNA干扰组细胞的穿膜细胞数（11.40个±1.36个）较阴性对照组（26.80个±1.83个）和空载体对照组（25.80个±1.93个）降低，差异有统计学意义（t=29.44，P=0.001；t=23.17，P=0.005）。细胞周期：RNA干扰组的S期细胞比例（28.26%±0.38%）显著低于空载体对照组（44.73%±0.66%）和阴性对照组（45.15%±1.69%），差异有统计学意义（t=63.69，P=0.000;t=71.55，P=0.000)，RNA干扰组的G2-M期细胞比例（26.99%±0.29%）显著高于阴性对照组（13.26%±0.49%）和空载体对照组（12.41%±0.46%），差异有统计学意义(t=56.31，P=0.000;t=51.39，P=0.000)。RNA干扰组细胞凋亡率（8.90%±0.48%）与阴性对照组（5.84%±0.21%）和空载体对照组（5.99%±0.37%）比较，差异均有统计学意义（t=51.34，P=0.000；t=47.27，P=0.000）。结论 S100A6基因参与肺腺癌细胞的增殖、浸润、细胞周期、细胞凋亡等生物学过程，与肿瘤发生、发展、转移等密切相关，有望作为肺腺癌诊断和治疗新的分子靶标。

关键词: 肺肿瘤, 细胞增殖, RNA干扰, 浸润, S100A6基因

Abstract: Objective To investigate the influence of S100A6 gene RNA interference on the biological behaviors of A549 lung adenocarcinoma cells. Methods The S100A6 gene RNA interference vector was transfected in A549 lung adenocarcinoma by lentivirus. The experiment was divided into three groups: pLenR-GPH group (the vector without S100A6 RNAi gene was transfected), negative control group (no vectors was transfected),  and  RNAi group (the vector with S100A6 RNAi gene was transfected). S100A6 mRNA and protein were detected using realtime PCR and Western blotting. The biological behavior including cell proliferation, invasion, cell cycle and cell apoptosis were detected by 3-(4, 5-dimethyl-2-thiazoly)-2, 5-diphenyl-2H-tetrazolium bromide, transwell, and flow cytometer, respectively. Results The expression of S100A6 mRNA of A549 lung adenocarcinoma cell line in RNAi group (0.009±0.001) was significantly decreased than those in negative control group (0.049±0.005) and pLenR-GPH group (0.030±0.006), with statistically significant differences (t=57.56, P=0.000; t=48.21, P=0.000). The expression of S100A6 protein of A549 lung adenocarcinoma cell line in RNAi group (0.107±0.002) was significantly decreased than those in negative control group (0.341±0.005) and pLenR-GPH group (0.311±0.006), with statistically significant differences (t=37.34, P=0.000; t=27.51, P=0.001). The ability of cell proliferation at 48 hours in RNAi group (0.230±0.008) was significantly declined than those in negative control group (0.292±0.038) and pLenRGPH group (0.307±0.013), with statistically significant differences (t=25.31, P=0.003; t=29.42, P=0.001). The number of transmembrane cells in RNAi group (11.40±1.36) was significantly declined than those in negative control group (26.80±1.83) and pLenRGPH group (25.80±1.93), with statistically significant differences (t=29.44, P=0.001; t=23.17, P=0.005). The cell proportion of S phase in RNAi group (28.26%±0.38%) was significantly lower than those in pLenR-GPH group (44.73%±0.66%) and negative control group (45.15%±1.69%)， with statistically significant (t=63.69, P=0.000; t=71.55, P=0.000). Cell propotion of G2-M phase in RNAi group (26.99%±0.29%) was significantly higher than those in negative control group (13.26%±0.49%) and pLenR-GPH group (12.41%±0.46%)， with statistically significant (t=56.31, P=0.000; t=51.39, P=0.000). The cell apoptosis proportion in RNAi group (8.90%±0.48%) was significantly higher than those in negative control group (5.84%±0.21%) and pLenRGPH group (5.99%±0.37%), with statistically significant (t=51.34, P=0.000; t=47.27, P=0.000). Conclusion S100A6 gene involves the proliferation, invasion, cell cycle and apoptosis of tumor cells, which has close correlation with occurrence, development and metastasis of lung adenocarcinoma. S100A6 gene is hopeful to become a molecular target for the diagnosis and treatment of lung adenocarcinoma.

Key words: Lung neoplasms, Cell proliferation, RNA interference, Invasion, S100A6 gene