miR-21 promotes cell migration and invasion of hepatocellular carcinoma by targeting KLF5 - PubMed (original) (raw)
miR-21 promotes cell migration and invasion of hepatocellular carcinoma by targeting KLF5
Jian Wang et al. Oncol Lett. 2019 Feb.
Abstract
Previous studies have reported that microRNAs regulate gene expression and transcription. miR-21 have been identified to play a role in many types of cancer. KLF5 functions as a tumor inhibitor in certain cancers. However, the role of KLF5 plays in hepatocellular carcinoma (HCC), especially concerning the relationship between miR-21 and the KLF5 gene remains to be determined. Reverse transcription-quantitative PCR (RT-qPCR), western blot analysis, as well as luciferase reporter and Transwell assays were used to determine the expression of miR-21 and KLF5 in Huh 7, SK-HEP-1, LO-2, and HCC tissues. In HCC cells and tissues, the upregulation of miR-21 was identified. HCC cell migratory and invasive abilities significantly increased because of miR-21 overexpression. KLF5 expression was inhibited by miR-21 by targeting its 3'-UTR. KLF5 overexpression alleviated the effect induced by miR-21 on the migratory and invasive ability of the Huh 7 cells. The results therefore show that, HCC cell migration and invasion is significantly suppressed by miR-21 via targeting KLF5. The newly identified miR-21/KLF5 axis provides a useful therapeutic biomarker for HCC treatment.
Keywords: Kruppel-like family of transcription factor 5; hepatocellular carcinoma; invasion; miR-21; migration.
Figures
Figure 1.
miR-21 was significantly upregulated in HCC tissues. (A) The expression levels of the miR-21 in HCC and theirs corresponding tissues were tested using RT-qPCR. The expression levels were calculated using the miR21/U6 expression ratio (2−∆∆Cq). (B) The miR-12 expression in HCC cell lines (Huh 7 and SK-HEP-1) and the LO-2 normal liver cells were tested using RT-qPCR. (C) The overall survival rate of the patients associated with the expression level of miR-21. **P<0.01, ***P<0.001.
Figure 2.
Overexpression of miR-21 promotes HCC cell line invasion and migration in vitro. (A) The successful overexpression of miR-21 in Huh 7 was confirmed by RT-qPCR. (B and C) Cell line invasion and migration were determined using the Transwell assays in the two HCC cell lines (Huh 7). *P<0.05, **P<0.01.
Figure 3.
The expression of KLF5 can be suppressed by miR-21, which binds to 3′-UTR. (A) The binding site of KLF for miR-21 was predicted using the online TargetScan tool. (B) Huh 7 cells were transfected with the Renilla luciferase psiCHECK-2 plasmid, and psiCHECK-2 plasmid which had either wide-type or mutant KLF5 3′-UTR, and the miR-21 mimic. (C) Huh 7 cell lines were transfected with NC or the miR-21 mimic. The SK-HEP-1 cell line was transfected with NC or LNA-miR-21. The expression of the KLF5 was tested by RT-qPCR. (D) The protein expression of KLF5 was determined using western blot analysis in the Huh 7 and SK-HEP-1 cell lines. *P<0.05.
Figure 4.
Overexpression of KLF5 alleviates the miR-21-driving promoting effect of cell line migration and invasion in vitro. (A and B) Transwell assays were used to determine the Huh 7 cell line migration and invasion following transfection with the miR-21 mimic and KLF5 without its 3′-UTR or control plasmid. *P<0.05, **P<0.01.
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