Abstract
BackgroundmiRNA-27a has been confirmed as an important regulator in carcinogenesis and other pathological processes. Whether and how it plays a role in the laryngeal carcinoma is unknown.MethodsMature miRNA-27a expression in laryngeal cancer was detected by qRT-PCR. Gain-of-function studies using mature miR-27a were performed to investigate cell proliferation and apoptosis in the Hep2 cells. In silico database analysis and luciferase reporter assay were applied to predict and validate the direct target, respectively. Loss-of-function assays were performed to investigate the functional significance of the miR-27a target gene. qRT-PCR and Western blot were used to evaluate mRNA and protein levels of the target, respectively.ResultsmiR-27a was significantly up-regulated in the laryngeal tumor tissues compared to the adjacent non-tumor tissues. In silico database analysis result revealed that PLK2 is a potential target of miR-27a. luciferase reporter assay result showed the direct inhibition of miR-27a on PLK2-3′UTR. In the cases with miR-27a up-regulation, PLK2 protein expression level was significantly lower in cancer tissues than that in the adjacent non-tumor tissues, which showed a negative correlation with miR-27a expression level. Both miR-27a and knockdown of PLK2 caused the increase of the cell viability and colony formation and inhibition of the late apoptosis in the Hep2 cell lines. Moreover, miR-27a but not PLK2 also repressed the early apoptosis in the Hep2 cells. Additionally, no alteration of the Hep2 cell cycle induced by miR-27a was detected.ConclusionsmiR-27a acts as an oncogene in laryngeal squamous cell carcinoma through down-regulation of PLK2 and may provide a novel clue into the potential mechanism of LSCC oncogenesis or serve as a useful biomarker in diagnosis and therapy in laryngeal cancer.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2407-14-678) contains supplementary material, which is available to authorized users.
Highlights
MiRNA-27a has been confirmed as an important regulator in carcinogenesis and other pathological processes
Results miR-27a is up-regulated in the human Laryngeal squamous cell carcinoma (LSCC) specimens and Hep2 cells We explored the expression of miR-27a in LSCC by qRT-PCR in 67 paired of fresh LSCC tissues and the paired adjacent tissues
Hep2 cells were silenced by small interfering RNA (siRNA)-PLK2 and the PLK2 protein was detected by Western blot. β-actin was used as internal control. (C) Effect of siRNA-PLK2 on the Hep2 cell proliferation measured by the MTT assay
Summary
MiRNA-27a has been confirmed as an important regulator in carcinogenesis and other pathological processes. Whether and how it plays a role in the laryngeal carcinoma is unknown. Laryngeal squamous cell carcinoma (LSCC) is one of the most common head and neck cancers in the world. Much work is focused on the identification of useful biologic and molecular markers in the diagnosis and therapy of LSCC [4,5]. MicroRNAs (miRNAs), a 20–23 nt functional RNA molecule, are a class of short non-coding RNAs and play important regulatory roles by sequence-specific base pairing on the 3′ untranslated region (3′-UTR) of target messenger RNAs (mRNAs), in promoting mRNA degradation or inhibiting translation [6]. It has been suggested that miRNA may be a molecular target for cancer diagnosis and therapy
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