Research Papers:

MACC1 is post-transcriptionally regulated by miR-218 in colorectal cancer

Katharina Ilm, Steffen Fuchs, Giridhar Mudduluru and Ulrike Stein _

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Oncotarget. 2016; 7:53443-53458. https://doi.org/10.18632/oncotarget.10803

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Katharina Ilm1, Steffen Fuchs1, Giridhar Mudduluru1,2, Ulrike Stein1,2

1Experimental and Clinical Research Center, Charité University Medicine Berlin and Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany

2German Cancer Consortium (DKTK), Heidelberg, Germany

Correspondence to:

Ulrike Stein, email: ustein@mdc-berlin.de

Giridhar Mudduluru, email: giridhar.mudduluru@mdc-berlin.de

Keywords: colorectal cancer, MACC1, miR-218, epigenetic regulation, alternative polyadenylation

Received: May 20, 2016     Accepted: July 13, 2016     Published: July 23, 2016


Metastasis is a multistep molecular network process, which is lethal for more than 90% of the cancer patients. Understanding the regulatory functions of metastasis-inducing molecules is in high demand for improved therapeutic cancer approaches. Thus, we studied the post-transcriptional regulation of the crucial carcinogenic and metastasis-mediating molecule metastasis associated in colon cancer 1 (MACC1). In silico analysis revealed MACC1 as a potential target of miR-218, a tumor suppressor miRNA. Expression of these two molecules inversely correlated in colorectal cancer (CRC) cell lines. In a cohort of CRC patient tissues (n = 59), miR-218 is significantly downregulated and MACC1 is upregulated compared with normal mucosa. Luciferase reporter assays with a construct of the MACC1-3′-UTR harboring either the wild type or the mutated miR-218 seed sequence confirmed the specificity of the targeting. miR-218 inhibited significantly MACC1 protein expression, and consistently, MACC1-mediated migration, invasion and colony formation in CRC cells. Anti-miR-218 enhanced the MACC1-mediated migration, invasion and colony formation. Similar findings were observed in the gastric cancer cell line MKN-45. Further, we performed methylation-specific PCR of the SLIT2 and SLIT3 promoter, where miR-218 is encoded in intronic regions. The SLIT2 and SLIT3 promoters are hypermethylated in CRC cell lines. miR-218 and SLIT2 expressions correlated positively. Methyltransferase inhibitor 5-Azacytidine induced miR-218 expression and inhibited the expression of its target MACC1. We also determined that MACC1 has alternative polyadenylation (APA) sites, which results in different lengths of 3′-UTR variants in a CRC cell line. Taken together, miR-218 is post-transcriptionally inhibiting the MACC1 expression and its metastasis-inducing abilities.

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