Abstract
Background: MiR-198 has been considered as an inhibitor of cell proliferation, invasion, migration and a promoter of apoptosis in most cancer cells, while its effect on non-cancer cells is poorly understood. Methods: The effect of miR-198 transfection on HaCaT cell proliferation was firstly detected using Cell Count Kit-8 and the cell cycle progression was analyzed by flow cytometry. Using bioinformatics analyses and luciferase assay, a new target of miR-198 was searched and identified. Then, the effect of the new target gene of miR-198 on cell proliferation and cell cycle was also detected. Results: Here we showed that miR-198 directly bound to the 3′-UTR of CCND2 mRNA, which was a key regulator in cell cycle progression. Overexpressed miR-198 repressed CCND2 expression at mRNA and protein levels and subsequently led to cell proliferation inhibition and cell cycle arrest in the G1 phase. Transfection ofSiCCND2 in HaCaT cells showed similar inhibitory effects on cell proliferation and cell cycle progression. Conclusion: In conclusion, we have identified that miR-198 inhibited HaCaT cell proliferation by directly targeting CCND2.
Highlights
MiRNAs are small non-coding RNAs ranging in length from 19 to 25 nucleotides, which regulate gene expression usually by targeting the 3′-UTR of mRNA for translation repression, degradation or both [1]
Ten putative mRNAs were unanimously predicted by the three algorithms (Figure 2A), and among which, CCND2 was of particular interest since previous studies have reported its important role in the cell cycle progress
To verify if miR-198 expression affects the expression of CCND2 at both mRNA and protein levels, the miR-198 mimic its negative control were transfected to HaCaT cells
Summary
MiRNAs are small non-coding RNAs ranging in length from 19 to 25 nucleotides, which regulate gene expression usually by targeting the 3′-UTR of mRNA for translation repression, degradation or both [1]. D-type cyclins are known to play critical roles in cell cycle progression [11]. Various studies on different cells have shown miR-26a [15], miR-302b, miR-497 [16], miR-133b [17], miR-1, miR-206, miR-29 [18], and miR-603 [19] could regulate cell proliferation by targeting CCND2. We show that miR-198 represses the proliferation of HaCaT cells, a keratinocyte cell line, by targeting cyclin D2. MiR-198 may be a key regulator in keratinocyte cell growth
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