[1]余　俊,刘彤鸥,李晓兰.小分子干扰RNA沉默黏着斑激酶基因对人宫颈癌Hela细胞生物学特征的影响[J].新乡医学院学报,2018,35(4):266-271.[doi:10.7683/xxyxyxb.2018.04.003]
　YU Jun,LIU Tong-ou,LI Xiao-lan.Effect of focal adhesion kinase gene silence by small interfering RNA on the biological characteristics of human cervical carcinoma Hela cells[J].Journal of Xinxiang Medical University,2018,35(4):266-271.[doi:10.7683/xxyxyxb.2018.04.003]
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小分子干扰RNA沉默黏着斑激酶基因对人宫颈癌Hela细胞生物学特征的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
35
期数:
2018年4
页码:
266-271
栏目:
基础研究
出版日期:
2018-04-05

文章信息/Info

Title:
Effect of focal adhesion kinase gene silence by small interfering RNA on the biological characteristics of human cervical carcinoma Hela cells
作者:
余　俊1; 刘彤鸥1; 李晓兰2
（1.湖北中医药大学第一临床学院　湖北省中医院妇产科，湖北　武汉　430061；2.宜昌市第二人民医院妇产科，湖北　宜昌　443000）
Author(s):
YU Jun1; LIU Tong-ou1; LI Xiao-lan2
（1.Department of Gynaecology and Obstetrics，the First Clinical College of Hubei University of Chinese Medicine，Hubei Provincial Hospital of Traditional Chinese Medicine，Wuhan 430061，Hubei Province，China；2. Department of Gynaecology and Obstetrics，the Second People′s Hospital of Yichang City，Yichang 443000，Hubei Province，China）
关键词:
宫颈癌; 黏着斑激酶; 小分子干扰RNA; 丝氨酸/苏氨酸激酶/丝裂原活化蛋白激酶/细胞外信号调节激酶; 信号通路
Keywords:
cervical cancer; focal adhesion kinase; small interfering RNA; serine/threonine kinase/mitogen-activated protein kinase/extracellular regulated protein kinase; signaling pathway
分类号:
R737.33
DOI:
10.7683/xxyxyxb.2018.04.003
文献标志码:
A
摘要:
目的　探讨小分子干扰RNA（siRNA）沉默黏着斑激酶（FAK）基因对人宫颈癌Hela细胞生物学特征的影响。方法　培养人宫颈癌Hela细胞，根据转染物的不同分为siRNA-FAK组、siRNA-阴性对照组和空白对照组，实时荧光定量聚合酶链反应技术检测细胞中FAK基因表达，噻唑蓝法检测细胞增殖能力，Transwell法检测细胞迁移和侵袭能力，Western blot法检测各组细胞中FAK、RAF原癌基因丝氨酸/苏氨酸激酶-1（Raf-1）、磷酸化-Raf-1（p-Raf-1）、丝裂原活化蛋白激酶1/2（MEK1/2）、磷酸化-MEF1/2（p-MEK1/2）、细胞外信号调节激酶（ERK）和磷酸化-ERK（p-ERK）蛋白表达。结果　空白对照组、siRNA-阴性对照组、siRNA-FAK组细胞中FAK mRNA相对表达量、细胞凋亡率、细胞迁移细胞数、侵袭细胞数、FAK、Raf-1、p-Raf-1、MEK1/2、p-MEK1/2、ERK和p-ERK蛋白相对表达量及细胞增殖能力比较差异均有统计学意义（P<0.05）。空白对照组与siRNA-阴性对照组细胞中FAK mRNA的相对表达量、细胞增殖能力、细胞凋亡率、迁移细胞数、侵袭细胞数以及FAK、Raf-1、p-Raf-1、MEK1/2、p-MEK1/2、ERK和p-ERK蛋白表达比较差异均无统计学意义（P>0.05）；siRNA-FAK组细胞中FAK mRNA相对表达量、细胞增殖能力、迁移细胞数、侵袭细胞数以及FAK、p-Raf-1、p-MEK1/2、p-ERK蛋白相对表达量均显著低于空白对照组和siRNA-阴性对照组（P<0.05），细胞凋亡率及Raf-1、MEK1/2和ERK蛋白相对表达量均显著高于空白对照组和siRNA-阴性对照组（P<0.05）。结论　特异性抑制人宫颈癌Hela细胞中FAK基因可减少细胞增殖，促进细胞凋亡，抑制细胞迁移和侵袭，其机制可能与抑制Raf/MEK/ERK信号通路有关。
Abstract:
Objective　To investigate the effect of focal adhesion kinase （FAK） gene silence by small interfering RNA（siRNA） on the biological characteristics of human cervical carcinoma Hela cells.Methods　The human cervical carcinoma Hela cells were cultured divided into siRNA-FAK group，siRNA-negative control group and blank control group according to the different transfectants. The expression of FAK gene in the Hela cells was detected by real-time fluorescence quantitative polymerase chain reaction. The cell proliferation was detected by methylthiazolyldiphenyl-tetrazolium assay. The cell migration and invasion were detected by Transwell method. The expressions of FAK，RAF proto-oncogene serine/threonine kinase -1（Raf-1），phosphorylated Raf-1（p-Raf-1），mitogen-activated protein kinase 1/2（MEK1/2），phosphorylated MEK1/2（p-MEK1/2），extracellular regulated protein kinase （ERK） and phosphorylated ERK （p-ERK） protein were detected by Western blot.Results　There was significant difference in the relative expression of FAK mRNA，FAK，Raf-1，p-Raf-1，MEK1/2，p-MEK1/2，ERK，p-ERK protein and the apoptosis rate，the number of migrating cells，the number of invasive cells,cell proliferation ability among the blank control group，the siRNA-negative control group and the siRNA-FAK group（P<0.05）.There was no significant difference in the relative expression of FAK mRNA，FAK，Raf-1，p-Raf-1，MEK1/2，p-MEK1/2，ERK，p-ERK protein and cell proliferation，apoptosis rate，the numbers of migrating cells，the numbers of invasive cells between the blank control group and the siRNA-negative control group（P>0.05）. The relative expression of FAK mRNA，FAK，p-Raf-1，p-MEK1/2，p-ERK protein and the cell proliferation，the number of migrating cells，the number of invasive cells in siRNA-FAK group were significantly lower than those in the blank control group and the siRNA-negative control group（P<0.05）. The apoptosis rate and the expression of Raf-1，MEK1/2 and ERK protein in siRNA-FAK group were significantly higher than those in the blank control group and the siRNA-negative control group（P<0.05）.Conclusion　Specific inhibition of FAK gene in human cervical cancer Hela cells can reduce cell proliferation，promote cell apoptosis，inhibit cell migration and invasion，and its mechanism may be related to the inhibition of Raf/MEK/ERK signaling pathway.

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更新日期/Last Update: 2018-04-05