S6K1沉默对宫颈癌细胞侵袭迁移及相关基因表达的影响

发布时间：2018-04-26 11:28

本文选题：核糖体蛋白S激酶 + 宫颈癌　；参考：《中国老年学杂志》2017年19期

【摘要】：目的探讨RNA干扰核糖体蛋白S6激酶1(S6K1)对人宫颈癌细胞Hela侵袭迁移及相关基因表达的影响。方法根据文献报道及siRNA设计原则合成靶向S6K1基因的siRNA,以阳离子脂质体Lipofectamine 2000为载体转染对数生长期Hela细胞(沉默组),同时设转染不针对任何基因随机序列的阴性对照组和转染试剂的空白对照组。转染48 h后采用实时定量聚合酶链反应(qRT-PCR)检测干扰效率,分别于转染24、48、72 h后采用WST法评价靶向沉默S6K1对Hela细胞增殖的影响,采用Transwell小室法检测经S6K1沉默后对Hela细胞侵袭迁移的影响,采用Western印迹法检测S6K1沉默后Hela细胞中人基质金属蛋白酶(MMP)-2、MMP-9和基质金属蛋白酶组织抑制因子(TIMP)-1、TIMP-2的蛋白水平。结果以空白对照组为参照(其S6K1 mRNA水平为1.000),阴性对照组的S6K1 mRNA水平为(1.027±0.034),差异无统计学意义(P0.05),而沉默组的S6K1 mRNA水平(0.158±0.012)低于其余两组(P0.05),相对于空白对照组的沉默率为(92.4±3.7)%。与空白对照组和阴性对照组比较,沉默组转染后的增殖率降低,迁移实验和侵袭实验中的穿膜细胞数均降低,MMP-2、MMP-9蛋白水平均降低,而TIMP-1、TIMP-2蛋白水平均升高(P0.05)。结论 RNA靶向沉默S6K1表达可抑制人宫颈癌Hela细胞侵袭迁移,可能与其抑制MMP-2/9表达及促进TIMP-1/2表达有关。
[Abstract]:Objective to investigate the effects of RNA interfering ribosomal protein S6 kinase 1 (S6K1) on Hela invasion and migration and related gene expression in human cervical cancer cells. Methods siRNAs targeting S6K1 gene were synthesized according to literature reports and siRNA design principles. Cationic liposome Lipofectamine 2000 was used as vector to transfect logarithmic growth phase Hela cells (silencing group). Irradiation group and blank control group of transfection reagent. The interference efficiency was detected by real-time quantitative polymerase chain reaction qRT-PCR 48 h after transfection, and the effect of targeted silencing S6K1 on the proliferation of Hela cells was evaluated by WST assay after transfection for 48 h. Transwell chamber assay was used to detect the effect of S6K1 silencing on the invasion and migration of Hela cells, and Western blot was used to detect the protein levels of human matrix metalloproteinase MMP-2MMP-2MMP-9 and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in Hela cells after S6K1 silencing. Results compared with the control group (the S6K1 mRNA level was 1.000g), the S6K1 mRNA level of the negative control group was 1.027 卤0.034 (P 0.05), but the S6K1 mRNA level of the silencing group (0.158 卤0.012) was lower than that of the other two groups (P 0.05), and the silencing rate was 92.4 卤3.7 in comparison with the blank control group. Compared with the blank control group and the negative control group, the proliferation rate of the silencing group decreased, and the number of transmembrane cells in migration and invasion experiments decreased, while the level of TIMP-1 and TIMP-2 protein increased (P 0.05). Conclusion RNA targeting silencing of S6K1 expression can inhibit the invasion and migration of human cervical cancer Hela cells, which may be related to the inhibition of MMP-2/9 expression and the promotion of TIMP-1/2 expression.
【作者单位】：承德医学院附属医院妇科;南京医科大学附属南京市妇幼保健院医学研究中心;
【基金】：国家自然科学基金(81302304)
【分类号】：R737.33

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