S6K2沉默对宫颈癌细胞增殖凋亡及PI3K／Akt／NF-κB通路的影响

临床肿瘤学杂志 ›› 2018, Vol. 23 ›› Issue (10): 886-892.

S6K2沉默对宫颈癌细胞增殖凋亡及PI3K／Akt／NF-κB通路的影响

067000 河北承德承德医学院附属医院妇科

收稿日期:2018-04-18 修回日期:2018-07-21 出版日期:2018-10-31 发布日期:2019-03-20
通讯作者: 张玉娟

Effects of S6K2 silencing on proliferation， apoptosis and PI3K／Akt／NF-κB signaling pathway in cervical cancer cells

Department of Gynaecology， the Affiliated Hospital of Chengde Medical College， Chengde 067000

Received:2018-04-18 Revised:2018-07-21 Online:2018-10-31 Published:2019-03-20
Contact: ZHANG Yujuan

摘要/Abstract

摘要：

目的探讨RNA干扰核糖体蛋白S6激酶2（S6K2）基因表达对宫颈癌细胞增殖、凋亡及磷脂酰肌醇3激酶（PI3K）／蛋白激酶B（Akt）／核因子κB（NF-κB）信号通路的影响。方法通过生物信息学方法—Oncomine（肿瘤微阵列数据库）分析宫颈癌组织中S6K2的表达情况，向人宫颈癌细胞系HeLa和Caski分别转染靶向S6K2的siRNA片段（siRNAS6K2）和对照随机序列（siRNA-Ctrl），采用实时荧光定量PCR（QPCR）检测各组转染48 h后的S6K2 mRNA水平，CCK-8法检测增殖情况，Annexin Ⅴ-FITC／PI双染流式细胞术检测细胞凋亡率，Western blotting检测PI3K／Akt通路相关蛋白（p-Akt、Akt、p-mTOR和mTOR）水平，免疫荧光法评估NF-κB p65的核转位情况。结果生物信息学分析3个与宫颈癌相关的数据集Biewenga、Scotto和Zhai中的数据，发现宫颈癌组织中S6K2相对表达量均高于正常宫颈组织（P＜0.05）。QPCR和Western blotting结果均显示转染siRNA-S6K2后的S6K2 mRNA和蛋白水平均较对照组显著降低（P＜0.05）。转染siRNA-S6K2的HeLa和Caski细胞的增殖活力、p-Akt和p-mTOR水平降低而凋亡率升高，与转染siRNA-Ctrl的细胞比较，差异有统计学意义（P＜0.05）。免疫荧光法检测结果显示，转染siRNA-S6K2的宫颈癌细胞的核内荧光量降低，与转染siRNA-Ctrl的细胞比较，差异有统计学意义（P＜0.05）。结论S6K2在宫颈癌组织中高表达，且参与了宫颈癌细胞的增殖和凋亡，在宫颈癌防治中有一定潜能。

关键词: font-family:宋体, 宫颈癌')">">宫颈癌, 核糖体蛋白S6激酶2, 增殖, 凋亡

Abstract:

Objective To investigate the effect of RNA interference （siRNA） on ribosomal protein S6 kinase 2 （S6K2） expression on proliferation and apoptosis of cervical cancer cells and phosphatidylinositol 3 kinase （PI3K）／protein kinase B （Akt）／nuclear factor-κB （NF-κB） signaling pathway. Methods The expression of S6K2 in human cervical cancer tissues was analyzed by Oncomine （tumor microarray database）. Human cervical cancer cell lines HeLa and Caski were transfected with siRNA targeting S6K2 （siRNA-S6K2） and control random sequence （siRNA-Ctrl）， respectively. Real-time fluorescence quantitative PCR （QPCR） was used to detect the mRNA levels of S6K2 in each group. CCK-8 method was used to detect the cell proliferation. Annexin V-FITC／PI doublestaining flow cytometry was used to detect the cell apoptotic rates. Western blotting was used to detect the levels of PI3K／Akt pathwayrelated proteins （p-Akt， Akt， p-mTOR and mTOR） 48 h after transfection. Immunofluorescence was used to evaluate the nuclear activity of NF-κB p65. Results Oncomine bioinformatics analysis showed that the relative expression of S6K2 in cervical cancer tissues was significantly higher than that in normal cervical tissues in three cervical cancerrelated datasets （Biewenga, Scotto and Zhai）（P＜0.05）. The results of QPCR and Western blotting showed that the mRNA and protein levels of S6K2 after transfection of siRNA-S6K2 were significantly lower than those of the control group （P＜0.05）. The proliferative activity and levels of p-Akt and p-mTOR in HeLa and Caski cells transfected with siRNA-S6K2 decreased while the apoptotic rate increased， which was significantly different from that of cells transfected with siRNA-Ctrl （P＜0.05）. Immunofluorescence assay showed that the intranuclear fluorescence of cervical cancer cells transfected with siRNA-S6K2 decreased compared with the transfected siRNA-Ctrl cells, and the difference was statistically significant（P＜0.05）. Conclusion S6K2 is highly expressed in cervical cancer tissues and participates in the proliferation and apoptosis of cervical cancer cells， so S6K2 has certain potential in the prevention and treatment of cervical cancer.

Key words: font-family:宋体, Cervical cancer">Cervical cancerfont-family:宋体, ')">">, Ribosomal protein S6 kinase 2, Proliferation, Apoptosis

中图分类号:

R737.33

. S6K2沉默对宫颈癌细胞增殖凋亡及PI3K／Akt／NF-κB通路的影响[J]. 临床肿瘤学杂志, 2018, 23(10): 886-892.

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