Abstract:

Objective To investigate whether re-expression of protocadherin 10(PCDH10) induced by 5-aza-2’-deoxycytidine (5-Aza-CdR) could affect the invasion and migration of MDA-MB-231 cells, and to explore the possible mechanism. Methods Human breast cancer cell line MDA-MB-231 was cultured in vitro. Control group and 5-Aza-CdR treatment group were set up. PCDH10 mRNA expression in MDA-MB-231 cell line was determined by reverse transcription-polymerase chain reaction (RT-PCR); Transwell chamber and wound healing assay were performed to measure the invasion and migration capacity of the cells, and protein expression of PCDH10, DNA methyltransferase(DNMT)3A, DNMT3B, nuclear factor(NF)-κB p65, matrix metalloproteinases(MMP)-2 and MMP-9 were detec Western blotting. Results 5-Aza-CdR could reverse the methylation status of PCDH10 gene in MDA-MB-231 cells in a dose-dependent manner. Re-expression of PCDH10 significantly inhibited cell invasion and migration capacity in vitro. Western blottoing analysis revealed that the expression of DNMT3A, DNMT3B, NF-κB p65, MMP-2 and MMP-9 in MDA-MB-231 cells were down-regulated after exposure to 5-Aza-CdR. Conclusion Re-expression of PCDH10 significantly inhibits MDA-MB-231 invasion and migration capacity. The inhibitory effect is characterized that 5-Aza-CdR treatment down-regulates DNMT3A and DNMT3B levels, recovers the expression of anti-oncogene PCDH10, further blocks the activation of NF-κB p65, resultsing in a decrease in the secretion of MMP-2 and MMP-9.

Key words: Breast cancer, Protocadherin 10, Methylation, Invasion, Migration, RT-PCR, Western blotting, Human