Simultaneously expressed miR-424 and miR-381 synergistically suppress the proliferation and survival of renal cancer cells—Cdc2 activity is up-regulated by targeting WEE1

Abstract

OBJECTIVES:MiRNAs are intrinsic RNAs that interfere with protein translation. Few studies on the synergistic effects of miRNAs have been reported. Both miR-424 and miR-381 have been individually reported to be involved in carcinogenesis. They share a common putative target, WEE1, which is described as an inhibitor of G2/M progression. Here, we studied the synergistic effects of miR-424 and miR-381 on renal cancer cells.METHODS:The viability of 786-O cells was analyzed after transfection with either a combination of miR-424 and miR-381 or each miRNA alone. We investigated cell cycle progression and apoptosis with flow cytometry. To confirm apoptosis and the abrogation of G2/M arrest, we determined the level of pHH3, which is an indicator of mitosis, and caspase-3/7 activity. The expression levels of WEE1, Cdc25, γH2AX, and Cdc2 were manipulated to investigate the roles of these proteins in the miRNA-induced anti-tumor effects. To verify that WEE1 was a direct target of both miR-424 and miR-381, we performed a dual luciferase reporter assay.RESULTS:We showed that the combination of these miRNAs synergistically inhibited proliferation, abrogated G2/M arrest, and induced apoptosis. This combination led to Cdc2 activation through WEE1 inhibition. This regulation was more effective when cells were treated with both miRNAs than with either miRNA alone, indicating synergy between these miRNAs. WEE1 was verified to be a direct target of each miRNA according to the luciferase reporter assay.CONCLUSIONS:These data clearly demonstrate that these two miRNAs might synergistically act as novel modulators of tumorigenesis by down-regulating WEE1 expression in renal cell cancer cells.

Suppression of MicroRNA 424 Levels by Human Papillomaviruses Is Necessary for Differentiation-Dependent Genome Amplification

“…It is also possible that additional targets of miR-424 besides CHK1 contribute to regulating stable HPV genome replication. This is consistent with our observation that Wee1, a kinase that regulates G 2 , is also controlled by miR-424 (30). Since miRNAs target a large spectrum of cellular genes, it is likely other miR-424 targets are also involved in regulating HPV replication.…”

“…The CIN612 cells stably expressing miR-424 exhibited reduced levels of both total CHK1 and p-CHK1. We also examined another miR-424 target, Wee1 (26,30), which controls the G 2 checkpoint, and found that increased expression of miR-424 similarly reduced Wee1 levels (Fig. 3E).…”

MicroRNA‑381 protects myocardial cell function in children and mice with viral myocarditis via targeting cyclooxygenase‑2 expression

“…miR-381 is closely related to MDR1 gene and plays an important role in multidrug resistance (24). Together with miR-424, miR-381 can target WEE1 gene to inhibit the activity of renal cancer Cdc2 cells (25). Moreover, miR-381 is closely related to lung adenocarcinoma (26).…”

MicroRNA Expression Patterns Involved in Amyloid Beta–Induced Retinal Degeneration

“…These viral proteins can trigger cancer-related genes and motivate several signaling pathways including genetic instability, inflammatory, and host immune responses. 56 These pathways, especially the proliferation 57,61-63 and apoptosis 58,63 pathways, were included in our model. Thirdly, as we made predictions with limited confirmed miRNA data, some possible pathways might be missing as an inevitable undesired result.…”