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miR-182 and miR-135b Mediate the Tumorigenesis and Invasiveness of Colorectal Cancer Cells via Targeting ST6GALNAC2 and PI3K/AKT Pathway

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Abstract

Background

Metastasis is a leading cause of cancer-related death including colorectal cancer (CRC). MicroRNAs are known to regulate cancer pathways and to be expressed aberrantly in cancer. Aberrant sialylation is closely associated with malignant phenotype of tumor cells, including invasiveness and metastasis.

Aim

This study aimed to investigate the association of miR-182 and miR-135b with proliferation and invasion by targeting sialyltransferase ST6GALNAC2 in CRC cells and explore the potential molecular mechanism.

Methods

We measured the levels of miR-182, miR-135b, and ST6GALNAC2 in a series of CRC cell lines and tissues using real-time PCR. Bioinformatics analysis and luciferase reporter assay were performed to test the direct binding of miR-182 and miR-135b to the target gene ST6GALNAC2. We also analyzed the possible role of miR-182/-135b on colony formation, wound healing, invasion, and tube formation.

Results

The expression of miR-182 and miR-135b was higher in tumor tissues compared to adjacent noncancerous tissues of CRC patients, as well as up-regulated in SW620 cells than in SW480 cells with different metastatic potential. By applying bioinformatics analysis and luciferase reporter assay, we identified ST6GALNAC2 as the direct target of miR-182/-135b. Furthermore, miR-182/-135b inhibited significantly ST6GALNAC2 expression, and consistently, ST6GALNAC2 mediated migration, adhesion, invasion, proliferation, and tumor angiogenesis in CRC cell lines. Additionally, PI3K/AKT signaling pathway was regulated by miR-182/135b, which was partially blocked by altered level of ST6GALNAC2 in CRC.

Conclusions

The miR-182/-135b/ST6GALNAC2/PI3K/AKT axis may serve as a predictive biomarker and a potential therapeutic target in CRC treatment.

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Abbreviations

3′UTR:

3′-untranslated region

CRC:

Colorectal cancer

miRNAs:

microRNAs

siRNA:

Small interfering RNA

STs:

Sialyltransferases

PI3K:

Phosphoinositide-3 kinase

PBS:

Phosphate-buffered saline

HUVECs:

Human umbilical vein endothelial cells

BSA:

Bovine serum albumin

RAF1:

RAF proto-oncogene serine/threonine-protein kinase

ST6GALNAC2:

GalNAc alpha-2,6-sialyltransferase 2

GCNT3:

Beta-1,6-N-acetylglucosaminyltransferase 3

GALNT7:

Polypeptide N-acetylgalactosaminyltransferase 7

FUT8:

Alpha-1,6 fucosyltransferase 8

ST3GAL5:

GMP-NeuAc: lactosylceramide alpha-2,3-sialyltransferase

ST3GAL6:

GMP-NeuAc: alpha-2,3-sialyltransferase

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Acknowledgements

This work was supported by Grants from National Natural Science Foundation of China (81472014, 81772277).

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Author notes

  1. Shihua Luo and Xiang Ren have contributed equally to this work.

Authors and Affiliations

  1. College of Laboratory Medicine, Dalian Medical University, Dalian, 116044, Liaoning Province, China

    Li Jia, Shihua Luo, Yang Li, Jialei Hu, Bing Liu, Lifen Zhao, Yujia Shan & Huimin Zhou

  2. College of Stomatology, Dalian Medical University, Dalian, 116044, Liaoning Province, China

    Xiang Ren

  3. Department of Traumatology, Shanghai Ruijin Hospital, Jiaotong University, Shanghai, 200025, China

    Shihua Luo

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  1. Li Jia
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Correspondence to Li Jia.

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Jia, L., Luo, S., Ren, X. et al. miR-182 and miR-135b Mediate the Tumorigenesis and Invasiveness of Colorectal Cancer Cells via Targeting ST6GALNAC2 and PI3K/AKT Pathway. Dig Dis Sci 62, 3447–3459 (2017). https://doi.org/10.1007/s10620-017-4755-z

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