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PURPOSE: Chemoresistance is a concern in ovarian cancer patients, in whom survival remains. MicroRNA, a novel class of small RNAs, have frequently been found to be dysregulated in human malignancies and to act as negative regulators of gene expression. This study aimed to explore the function of miR-338-3p in cisplatin resistance in ovarian cancer and potential molecular mechanisms thereof. MATERIALS AND METHODS: The expression levels of miR-338-3p and WNT2B in ovarian cancer tissues and cells were estimated by real-time quantitative polymerase chain reaction (RT-qPCR). In addition, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazol-3-ium bromide (MTT), transwell, and flow cytometry assays were used to assess biological role of miR-338-3p in vitro. Western blot assay was conducted to measure protein expression of WNT2B, epithelial-mesenchymal transition (EMT)-related proteins, and apoptosis-related proteins. The relationship between miR-338-3p and WNT2B was confirmed by dual-luciferase reporter. Finally, a xenograft tumor model was developed to explore the effects of overexpression of miR-338-3p on tumor growth in ovarian cancer in vivo. RESULTS: MiR-338-3p was downregulated in cisplatin resistant ovarian cancer tissues and cells. Mechanistically, high expression of miR-338-3p enhanced cell sensitivity to cisplatin by inhibiting proliferation, motility, and EMT and by promoting apoptosis via targeting WNT2B expression in vitro. Furthermore, overexpression of miR-338-3p increased cisplatin sensitivity among ovarian cancer in an in vivo xenograft tumor model. CONCLUSION: MiR-338-3p enhances the sensitivity of ovarian cancer cells to cisplatin by downregulating WNT2B.
Subject(s)
Humans , Apoptosis , Blotting, Western , Cisplatin , Epithelial-Mesenchymal Transition , Flow Cytometry , Gene Expression , Heterografts , In Vitro Techniques , MicroRNAs , Ovarian Neoplasms , Polymerase Chain Reaction , RNAABSTRACT
Objective To investigate the role of miR-338-5p/SIRT1-related signaling pathway in the treatment of colorectal cancer by the chemopreventive effects of black raspberry (BRB) anthocyanins. Methods Mice were divided into normal healthy control group, AOM/DSS-induced colorectal cancer groups with or without BRB anthocyanin. miRNA microarray was used to investigate differentially expressed miRNAs and RT-qPCR was applied to verify the expression of selected miR-338-5p/SIRT1 in colon tissue of mice and human colorectal cancer cell lines of Caco-2, LoVo, HCT-116, HT29, and SW480. TargetScan and miRanda bioinformatics software was used to predict the targeted regulation relationships between miR-338-5p and SIRT1. The expression of SIRT1 protein in colon tissue of mice and downstream signaling pathway-related proteins were determined by Western blotting. Results miRNA microarray differential analysis demonstrated that the expression of miR-338-5p was significantly reduced in colon tissues of AOM/DSS induced mice fed with BRB anthocyanin. While after 9 weeks administration of BRB anthocyanins, the level of miR- 338-5p in AOM/DSS induced mice was decreased. The expression pattern of miR-338-5p was confirmed in the colon tissue and several colon cancer cell lines. Meanwhile, colorectal cancer cells were treated with BRB anthocyanin, miR-338-5p expression was reduced. TargetScan and miRanda predicted that the SIRT1 was one of target genes of miR-338-5p. BRB anthocyanins could promote the expression of SIRT1 protein in intestinal epithelial cells and regulate the protein levels of downstream moleculars including mTOR et al. Conclusion miR-338-5p/SIRT1-related signaling pathway might involve in the chemoprevention effects of BRB anthocyanin on colorectal cancer, which provided a new strategy for chemoprevention of colorectal cancer.
ABSTRACT
MicroRNAs(miRNAs) are a class of highly conserved,endogenous,small,non-coding RNAs that reg-ulate RNAi and are key regulators of tumorigenesis and development.miR-338 is abnormally expressed in most tumors. Aberrant expression of miR-338 can affect the proliferation, invasion, metastasis, angiogenesis and other physiological activities of tumor cells and play a vital role in the regulation on one or more target-genes.
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OBJECTIVES: MicroRNAs are important modulators of the cellular epithelial‑mesenchymal transition (EMT) process and are associated with metastasis in human nonsmall cell lung cancer (NSCLC). In this study, we tried to investigate the role of miR‑338‑3p in NSCLC cells. MATERIALS AND METHODS: Real‑time polymerase chain reaction was applied to quantify the expression levels of miR‑338‑3p, as well as EMT‑associated molecules in NSCLC cells. Wound healing and transwell assays were performed to evaluate the migration and invasion capacities, respectively. Dual‑luciferase reporter assay was finally performed to determine the targeting of zinc finger E‑box‑binding protein 2 (ZEB2) by miR‑338‑3p. RESULTS: We found that miR‑338‑3p was significantly reduced in NSCLC cell lines. Forced expression of miR‑338‑3p in A549 cells led to the suppression of migration/invasion capacity and inhibition of epithelial markers. In addition, we proved that miR‑338‑3p could directly target ZEB2. CONCLUSIONS: In general, we summarized that miR‑338‑3p could inhibit EMT and metastasis of human NSCLC cells, which probably via directly targeting ZEB2 expression.
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To explore the serum miR-338-5p expression characteristics in renal transplant recipients ,and the role of regulating BAFF signal ,then investigate its biological significance.Methods:Healthy volunteers were enrolled as control group.Serum miR-338-5p was detected by real-time PCR;soluble BAFF was detected by ELISA;anti-HLA-Ⅰantibody,anti-HLA-Ⅱ antibody and anti-MICA antibody were detected by liquid chip technology.SPSS17.0 software was applied.t-test was used to compare the means of two independent samples;Paired samples t-test was used to compare the means of two paired samples;Spearman method and Pearson method were used to analyse the correlation;P<0.05 was considered to be statistically significant.Results: Compared with healthy controls,serum miR-338-5p in renal transplant recipients decreased significantly (P<0.001),while serum BAFF increased significantly (P<0.01).Serum miR-338-5p levels within 1 year post-transplantation were significantly lower than that of more than 1 year post-transplantation (P<0.01);Serum miR-338-5p levels within 3 years post-transplantation were significantly lower than that of more than 3 years post-transplantation (P<0.01);To all research objects,serum miR-338-5p was significantly negatively correlated with serum BAFF (r=-0.51,P<0.001),and serum miR-338-5p was significantly negatively correlated with anti-HLA-Ⅱ antibody(r=-0.322, P<0.05);Serum miR-338-5p within 3 years was significantly negatively correlated with anti-HLA-Ⅱantibody (r=-0.423,P<0.05), and serum miR-338-5p within 3 years was significantly negatively correlated with anti-MICA antibody(r=-0.411,P<0.05);Serum miR-338-5p more than 3 years was significantly positively correlated with anti-MICA antibody(r=0.486,P<0.05),and Serum miR-338-5p more than 3 years was significantly positively correlated with anti-HLA & MICA antibody(r=0.578,P<0.01).Conclusion:miR-338-5p may directly or indirectly target BAFF signal ,and participate antibody mediated immune response by regulating its target genes and interfere with the long-term survival of transplanted renal.