Role of microRNA-21 in uveal melanoma cell invasion and metastasis by regulating p53 and its downstream protein

International Journal of Ophthalmology
Ying-Chih WangXiao-Lin Xu

Abstract

To reveal the insight mechanism of liver metastasis in uveal melanoma, we investigated cell functions of microRNA-21 in three different uveal melanoma cell lines and analyze the relationship of target gene p53 and its downstream targets. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect microRNA-21 expression in normal uveal tissue and uveal melanoma cell lines. Lenti-virus expression system was used to construct OCM-1, MuM-2B and M619 cell line with stable overexpression and inhibition of microRNA-21. In vitro cell function tests such as cell proliferation, cell apoptosis, cell circle and abilities of migration and invasion were examined by MTT, BrdU assay, flow cytometry, transwell assay and Matrigel invasion assay respectively. The target gene was predicted by bioinformatics and confirmed by using a dual luciferase reporter assay. The expression of p53 and its suspected downstream targets LIM and SH3 protein 1 (LASP1) and glutathione S transferase pi (GST-Pi) were determined by qRT-PCR in mRNA level and Western blotting analysis in protein level. Finally, the effect of microRNA-21 in a xenograft tumor model was assessed in four-week-old BALB/c nude mice. Compared to normal uveal melano...Continue Reading

Citations

Aug 13, 2020·Cellular and Molecular Life Sciences : CMLS·Chun YangPierre Hardy
Jul 17, 2020·OncoTargets and Therapy·Yi Fan LiWen Bin Wei
May 11, 2021·Frontiers in Oncology·Soudeh Ghafouri-FardMohammad Taheri

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