Department of Histology and Embryology, Basic Medical College of Zhengzhou University, Zhengzhou, Henan, China.
Eur Rev Med Pharmacol Sci. 2018 Apr;22(8):2287-2296. doi: 10.26355/eurrev_201804_14817.
To investigate the role of miR-198 and its target gene Toll-like receptor 4 (TLR4) of tumorigenesis of gastric cancer (GC).
The expression of miR-198 in GC cells was detected by quantitative polymerase chain reaction (qPCR). The proliferation, apoptosis, migration, and invasion of GC cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), flow cytometry, transwell chamber, and wound scratch assay. Bioinformatics analysis for the results of protein chip was performed to identify the target genes of miR-198. TLR4 was further confirmed to be the target gene of miR-198 by TLR4 luciferase reporter assay.
miR-198 expression level in GC SGC-7901 cells significantly decreased compared with the normal cells. When the miR-198 was overexpressed, the proliferation, migration, and invasion of GC cells were significantly decreased, while the apoptosis was increased. The expression of TLR4 in SGC-7901 cells was significantly higher, while the expression of TLR4 in SGC-7901 cells transfected with miR-198 significantly lowered, which was consistent with the Western blot for TLR4. The luciferase reporter assay confirmed that TLR4 was the target genes of miR-198 in GC SGC7901 cells.
miR-198 could induce apoptosis and inhibit the proliferation, migration, and invasion of GC cells through downregulating TLR4 expression.
探讨微小 RNA-198(miR-198)及其靶基因 Toll 样受体 4(TLR4)在胃癌(GC)发生中的作用。
采用实时定量聚合酶链反应(qPCR)检测 GC 细胞中 miR-198 的表达。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)、流式细胞术、Transwell 小室和划痕实验检测 GC 细胞的增殖、凋亡、迁移和侵袭。通过蛋白质芯片结果的生物信息学分析鉴定 miR-198 的靶基因。进一步通过 TLR4 荧光素酶报告基因实验验证 TLR4 是 miR-198 的靶基因。
与正常细胞相比,GC SGC-7901 细胞中 miR-198 的表达水平明显降低。当 miR-198 过表达时,GC 细胞的增殖、迁移和侵袭明显减少,而凋亡增加。SGC-7901 细胞中 TLR4 的表达明显升高,而转染 miR-198 的 SGC-7901 细胞中 TLR4 的表达明显降低,这与 TLR4 的 Western blot 结果一致。荧光素酶报告基因实验证实 TLR4 是 GC SGC7901 细胞中 miR-198 的靶基因。
miR-198 可通过下调 TLR4 的表达诱导 GC 细胞凋亡,并抑制其增殖、迁移和侵袭。