ABSTRACT
BACKGROUND: Non-coding RNAs have attracted considerable attention for their vital role in cancer. The purpose of this study was to determine the effects of non-coding RNAs on hepatocellular carcinoma (HCC) and reveal their regulatory mechanism in the pathophysiological process. METHODS: We measured the expression of mucin 1 (MUC1) and miR-485-5p in tissues from 15 HCC patients and in liver cancer cell lines by quantitative real-time polymerase chain reaction and Western blot, screened for aberrantly expressed microRNAs (miRNAs) by miRNA microarrays. Bioinformatics tools were used to find the miRNA and circular RNA that regulated MUC1, which were validated by RNA immunoprecipitation assay and luciferase reporter assay. Cell counting kit-8, Transwell assays, and flow cytometry were used to conduct functional experiments. Proteins were examined by western blot and immunohistochemical staining assays. Significant differences between groups were estimated using the one-way analysis of variance. A Pâ<â0.05 was considered statistically significant. RESULTS: MUC1 was overexpressed in HCC tissues compared with that in paratumor tissues (normal vs. tumor, 1.007â±â0.215 vs. 75.213â±â18.403, t = 18.401, P < 0.001) while miR-485-5p was down-regulated (normal vs. tumor, 4.894â±â0.684 vs. 1.586â±â0.398, tâ=â16.191, P < 0.001). Inhibition of miR-485-5p promoted cell proliferation (73.33%â±â5.13% vs. 41.33%â±â3.51%, tâ=â8.913, P < 0.001), migration (102â±â8 cells vs. 46â±â8 cells, tâ=â8.681, P < 0.001), invasion (59â±â7 cells vs. 28â±â2 cells, tâ=â8.034, P < 0.01), and suppressed apoptosis (22.64%â±â6.97% vs. 36.33%â±â3.96%, tâ=â2.958, P < 0.05) of HepG2 cells with which MUC1 is knocked down. Mechanically, miR-485-5p binds to MUC1, while circHECTD1 binds to miR-485-5p, resulting in the indirect up-regulation of the MUC1 level. CONCLUSIONS: Our findings reveal that circHECTD1 facilitates HCC progression by sponging miR-485-5p to up-regulate MUC1.
Subject(s)
Carcinoma, Hepatocellular , Liver Neoplasms , MicroRNAs , Mucin-1 , Carcinoma, Hepatocellular/genetics , Cell Line, Tumor , Cell Proliferation/genetics , Gene Expression Regulation, Neoplastic , Humans , Liver Neoplasms/genetics , MicroRNAs/genetics , Mucin-1/genetics , RNA, Circular , Ubiquitin-Protein LigasesABSTRACT
Gastric cancer stem cells (GCSCs) have an important role in metastasis and recurrence of gastric cancer, and novel treatment strategies that target GCSCs are urgently required. Although evodiamine (Evo), a derivative of the traditional herbal medicine Evodia rutaecarpa, has been reported to have various biological effects, its effect on GCSCs remains unknown. In order to determine the effect of Evo on apoptosis of GCSCs, an MTS assay, flow cytometry and western blot analysis were performed. The effect of Evo on selfrenewal in GCSCs was measured by alterations in the sphere formation ability, the expression of inducedpluripotent stem cell factors, expression of epithelial-to-mesenchymal transition (EMT) factors and oxaliplatin resistance of gastric cancer cells (GCCs). Evo inhibited proliferation, promoted the Bax/Bcell lymphoma 2 ratio and altered active caspase3 expression of GCSCs. In addition, Evo decreased the sphere formation ability, the expression of Sox2, KLF4, Bmi1 and Oct4, and oxaliplatin resistance in GCCs. Evo decreased the expression of Slug, Twist, Zeb1 and vimentin, suggesting an inhibitory effect on EMT. Furthermore, the expression of ßcatenin, cMyc and cyclin D1 was decreased in Evotreated spheroids from GCCs. In conclusion, Evo inhibited the Wnt/ßcatenin signaling pathway to inhibit proliferation and stem cell properties of GCSCs and repressed the EMT. The present findings highlight the prospect of Evo as a CSCs-targeted therapy in gastric cancer.
Subject(s)
Cell Proliferation/drug effects , Neoplastic Stem Cells/drug effects , Quinazolines/pharmacology , Wnt Signaling Pathway/drug effects , Animals , Apoptosis/drug effects , Blotting, Western , Cell Line, Tumor , Cell Survival/drug effects , Cyclin D1/metabolism , Epithelial-Mesenchymal Transition/drug effects , Flow Cytometry , Humans , Kruppel-Like Factor 4 , Molecular Structure , Neoplastic Stem Cells/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Proto-Oncogene Proteins c-myc/metabolism , Quinazolines/chemistry , Snail Family Transcription Factors , Stomach Neoplasms/metabolism , Stomach Neoplasms/pathology , Transcription Factors/metabolism , Twist-Related Protein 1/metabolism , bcl-2-Associated X Protein/metabolism , beta Catenin/metabolismABSTRACT
Nuclear receptors are a superfamily of transcription factors including the steroid hormone receptors, non-steroid hormone receptors and the orphan nuclear receptor family. Retinoic acid-related orphan receptor (ROR)ß, as a member of the orphan nuclear receptor family, plays an important regulatory role in the maintenance of a variety of physiological and pathological processes. RORß has been determined to act as an osteogenic repressor in regulating bone formation, and is involved in regulating circadian rhythm. The findings of recent studies concerning the association between tumorigenesis and circadian rhythm have shown that an aberrant circadian rhythm may promote tumorigenesis and tumor progression. The mechanisms discussed in this review demonstrate how aberrant RORß-induced circadian rhythm may become a new direction for future studies on tumorigenesis and strategy design for cancer prevention.
