MiR-23a transcriptional activated by Runx2 increases metastatic potential of mouse hepatoma cell via directly targeting Mgat3

Abstract

MicroRNAs (miRNAs) and aberrant glycosylation both play important roles in tumor metastasis. In this study, the role of miR-23a in N-glycosylation and the metastasis of mouse hepatocellular carcinoma (HCC) cells was investigated. The miRNA expression array profiles that were confirmed by qPCR and Western blot analyses revealed higher miR-23a expression levels in Hca-P cells (with lymphatic metastasis potential) than in Hepa1–6 cells (with no lymphatic metastasis potential), while the expression of mannoside acetylglucosaminyltransferase 3 (Mgat3) was negatively associated with metastasis potential. Mgat3 is a key glycosyltransferase in the synthesis of the bisecting (β1,4GlcNAc branching) N-glycan structure. Bioinformatics analysis indicated that Mgat3 may be a target of miR-23a, and this hypothesis was verified by dual-luciferase reporter gene assays. Furthermore, we found that the transcription factor Runx2 can directly bind to the miR-23a gene promoter and promote its expression, as shown in dual-luciferase reporter gene assays and ChIP assays. Collectively, these results indicate that miR-23a might increase the metastatic potential of mouse HCC by affecting the branch formation of N-glycan chains presented on the cell surface through the targeting of the glycosyltransferase Mgat3. These findings may provide insight into the relationship between abnormal miRNA expression and aberrant glycosylation during tumor lymphatic metastasis.

RETRACTED ARTICLE: Caveolin-1 promotes invasion and metastasis by upregulating Pofut1 expression in mouse hepatocellular carcinoma

“…Total protein extracts and western blot procedures were carried out as previously described 25 . Antibodies specific for the following targets were used: HNF4A (Abcam, USA); Sp1, c-Myc, and CREB (Cell Signaling Technology, USA); p-HNF4A (Ser313), p-Sp1 (1:500, Thr739), p-c-Myc (Ser62), p-CREB (Ser133), p-JNK (Thr183+Tyr185), and p-p38 (Tyr323) (Biosynthesis Biotechnology, Beijing, China); extracellular signal–regulated kinase (ERK; Proteintech, Wuhan, China); p-ERK (Thr202+Tyr204, Bioworld, Nanjing, China); and c-Jun N-terminal kinase (JNK), p38, and GAPDH (Proteintech, Wuhan, China).…”

Caveolin‐1 upregulates Fut8 expression by activating the Wnt/β‐catenin pathway to enhance HCC cell proliferative and invasive ability

“…The reverse transcription experiment followed the procedure of PrimeScript™ RT Reagent Kit (TaKaRa, Japan). qPCR was carried out as previously described (Huang et al, 2018). The upstream and downstream primers sequences for Cav‐1, Fut8, β‐catenin, Gapdh , and β‐actin are shown in Table 1.…”

Reduced expression of ppGalNAc‐T4 promotes proliferation of human breast cancer cells

“…Total protein extracts, western blot and lectin blot were performed as previously described (Huang et al, 2018). The following antibodies and lectins were obtained: anti‐GAPDH, anti‐phosphorylated retinoblastoma protein (p‐Rb), anti‐Cyclin D1, anti‐CDK1, anti‐Cyclin E2, anti‐p57, anti‐p27, anti‐Cyclin A1/2, Cullin antibodies from CST, anti‐Notch1, Hes‐related with YRPW motif (Hey1), hairy/enhancer‐of‐split (Hes1) antibodies from Proteintech, biotinylated Vicia villosa lectin (VVL) from Vector Laboratories, respectively.…”