The lncRNA TDRG1 promotes cell proliferation, migration and invasion by targeting miR-326 to regulate MAPK1 expression in cervical cancer.

BACKGROUND: Recently, lncRNA-Testis developmental related gene 1 (TDRG1) was proved to be a key modulator in reproductive organ-related cancers. The biological role of TDRG1 in cervical cancer (CC) progression remains largely unknown. METHOD: Real-time PCR (qRT-PCR) examined the expression level of TDRG1, microRNA (miR)-326 and MAPK1 mRNA. OS tissues and corresponding relative normal tissues, as well as CC cell lines and normal cell line Ect1/E6E7 were collected to determine the expression of TDRG1 in CC. MTT, colony formation, wound-healing, transwell and flow cytometer assay detected the influence of TDRG1 and miR-326 on CC cells growth, metastasis and apoptosis. Western blot examined proteins level. Bioinformatics, RNA pull-down assay, RNA immunoprecipitation and dual-luciferase reporter assays detected the molecular mechanism of TDRG1 in CC. Xenograft tumour model was established to determine the role of TDRG1 in vivo. RESULTS: The expression of TDRG1 was significantly increased in CC tissues and cell lines compared with normal tissue and normal cell line respectively and its expression was associated with clinicopathological characteristics of CC patients. Knockdown of TDRG1 inhibited the cell proliferation, migration and invasion in Hela and SIHA cells. Moreover, TDRG1 directly interacted with miR-326, and the inhibition effect on cell growth and metastasis induced by TDRG1 siRNA can be abrogated by miR-326 silencing by its inhibitor in Hela and SIHA cells. Further, MAPK1 was proved to be a direct target of miR-326, and its expression was negatively regulated by miR-326 while positively modulated by TDRG1. CONCLUSION: TDRG1 acts as a competing endogenous lncRNA (ceRNA) to modulate MAPK1 by sponging miR-326 in CC, shedding new light on TDRG1-directed diagnostics and therapeutics in CC.

Knockdown of ACTA2­AS1 promotes liver cancer cell proliferation, migration and invasion.

Long noncoding RNAs (lncRNAs) are important regulators of various cellular and biological processes. The present study aimed to investigate the functions of a novel lncRNA, ACTA2­AS1:4, a transcript variant of smooth muscle α­actin 2­antisense 1 (ACTA2­AS1), in regulating liver cancer progression. Expression of lncRNAs in liver cancer tissues and cell lines were analyzed by reverse transcription quantitative polymerase chain reaction (RT­qPCR). Knockdown of ACTA2­AS1:4 expression in LM3 liver cancer cells was achieved by transfection with small interfering RNAs (siRNAs) that specifically targeted ACTA2­AS1:4. The proliferation and cell cycle progression of ACTA2­AS1:4­silenced LM3 cells were determined using MTS assay and flow cytometry, respectively. A Transwell system assay was used to evaluate the migration and invasion capacities of LM3 cells transfected with ACTA2­AS1:4 siRNA. The expression levels of major genes associated with important cellular processes were finally determined by RT­qPCR and western blot analysis. ACTA2­AS1:4 expression in liver cancer tissues and multiple cell lines was markedly downregulated by specific siRNAs. This inhibition of ACTA2­AS1:4 expression significantly promoted the proliferation, cell cycle progression, migration and invasion of LM3 cells. A decrease in ACTA2­AS1:4 expression also suppressed E­cadherin expression, increased N­cadherin expression, decreased caspase 3 expression and increased cyclin D1 and matrix metalloproteinase expression in liver cancer cells. Downregulation of ACTA2­AS1:4 affects a number of key mechanisms involved in liver cancer progression. These data may be important for the future of liver cancer diagnosis and subsequent treatments.

MicroRNA-122a Regulates Zonulin by Targeting EGFR in Intestinal Epithelial Dysfunction.

BACKGROUND/AIMS: This study aimed to investigate the role of microRNA (miR)-122a in regulating zonulin during the modulation of intestinal barrier. METHODS: Zonulin proteins and their target gene expression were analyzed in miR-122a-overexpressing cell lines and in the target gene of epidermal growth factor receptor (EGFR). An mmu-miR-122a intestinal epithelial conditional transgenic (miR-122a-TG) mouse model was established to investigate EGFR and zonulin expression. MiR-122a was also detected in the clinical specimens of inflammatory bowel disease. RESULTS: EGFR was identified as a target gene of miR-122a. The expression level of miR-122a was positively correlated with that of zonulin. The expression level of zonulin was significantly increased, whereas the expression level of EGFR was significantly decreased in the miR-122a-TG mice and in the corresponding primary epithelial culture (P < 0.05). These results were consistent with the data of the clinical specimens. CONCLUSIONS: miR-122a could be a positive factor of zonulin by targeting EGFR, which increased the intestinal epithelial permeability in vivo and in vitro.

Long non-coding RNA LINC00673 promotes hepatocellular carcinoma progression and metastasis through negatively regulating miR-205.

Increasing evidence demonstrates abnormal expression of long non-coding RNA (lncRNA) is closely correlated with various malignancies including hepatocellular carcinoma (HCC). The present study aims to investigate the role of lncRNA long intergenic noncoding RNA 00673 (LINC00673) in tumorigenesis of HCC. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed LINC00673 was upregulated in HCC cancerous tissue and cell lines compared to adjacent normal tissue and normal liver cell lines. LINC00673 overexpression is associated with poor prognosis and low survival rate. LINC00673 silencing inhibited the proliferation, invasion and epithelial-mesenchymal transition (EMT) of HCC cells in vitro. Bioinformatics analysis revealed that miR-205 targeted 3'-UTR of LINC00673. Rescue experiments confirmed that miR-205 could reverse the effect of LINC00673 on HCC cells. In vivo xenograft tumor assay LINC00673 silencing reduced the tumor volume and weight. Taken together, findings indicate overexpression of LINC00673 promotes HCC cells progression by regulating miR-205, providing a prognostic biomarker and therapeutic target for HCC and is associated with poor survival of HCC patients.