Research Paper Volume 13, Issue 4 pp 4962—4975

Methylation-dependent MCM6 repression induced by LINC00472 inhibits triple-negative breast cancer metastasis by disturbing the MEK/ERK signaling pathway

LINC00472 down-regulates MCM6 expression via recruitment of DNA methyltransferases. (A) The subcellular localization of LINC00472 predicted on lncATLAS. (B) The subcellular localization (× 400) of LINC00472 detected by FISH assay. (C) The nuclear and cytoplasmic expression of LINC00472 in MDA-MB-231 cells as determined by means of RT-qPCR. (D) The binding relation between LINC00472 and DNMT1, DNMT3a or DNMT3b analyzed on RPISeq, with Random Forest and Support Vector Machine > 0.5 indicative of positive results. (E) The binding relation between LINC00472 and DNMT1, DNMT3a or DNMT3b relative to IgG detected by means of RIP assay. (F) The expression of DNMT1, DNMT3a or DNMT3b pulled down by LINC00472 normalized to Input. (G) The enrichment of DNMT1, DNMT3a and DNMT3b in MCM6 promoter region normalized to IgG detected by means of ChIP assay. * p vs. the oe-NC group (MDA-MB-231 cells treated with oe-NC). The results were measurement data and expressed as mean ± standard deviation. Data comparison between two groups was analyzed by the independent sample t-test. The experiment was conducted 3 times independently.

Figure 4. LINC00472 down-regulates MCM6 expression via recruitment of DNA methyltransferases. (A) The subcellular localization of LINC00472 predicted on lncATLAS. (B) The subcellular localization (× 400) of LINC00472 detected by FISH assay. (C) The nuclear and cytoplasmic expression of LINC00472 in MDA-MB-231 cells as determined by means of RT-qPCR. (D) The binding relation between LINC00472 and DNMT1, DNMT3a or DNMT3b analyzed on RPISeq, with Random Forest and Support Vector Machine > 0.5 indicative of positive results. (E) The binding relation between LINC00472 and DNMT1, DNMT3a or DNMT3b relative to IgG detected by means of RIP assay. (F) The expression of DNMT1, DNMT3a or DNMT3b pulled down by LINC00472 normalized to Input. (G) The enrichment of DNMT1, DNMT3a and DNMT3b in MCM6 promoter region normalized to IgG detected by means of ChIP assay. * p < 0.05 vs. the oe-NC group (MDA-MB-231 cells treated with oe-NC). The results were measurement data and expressed as mean ± standard deviation. Data comparison between two groups was analyzed by the independent sample t-test. The experiment was conducted 3 times independently.