Epigenetic activation of alpha4, beta2 and beta6 integrins involved in cell migration in trichostatin A-treated Hep3B cells.
The epigenetic modulation by histone deacetylase (HDAC) inhibitors including trichostatin A (TSA) has been known to block cell proliferation, induce apoptosis and inhibit cell migration in human cancer cells that represents the potential therapeutic agents for cancers and fibrosis. However, more than 55% of Hep3B cells remained alive after our initial study of 100 nM TSA treatment. To further study the epigenetic modulation and the biological function of newly activated genes by HDAC inhibitor involved in HCC progression and metastasis, we profiled 23 integrin genes including 15alpha and 8beta in TSA-treated Hep3B cells. Six integrins including three down-regulated alpha6, alpha10, beta8 and three significant up-regulated alpha4, beta2, beta6 integrins were revealed after semi-quantitative RT-PCR. To confirm the epigenetic modulation and explore their biological functions, we selected the three significantly up-regulated integrins for confirmation of protein up-regulation, hyperacetylated-histones by ChIP assays, and functional inhibition by specific neutralizing antibodies of integrins. Our results indicated that epigenetic modulation in TSA-treated Hep3B cells up-regulated new integrins including alpha4, beta2 and beta6 and reduced migration activities by specific neutralizing antibodies to 61.3%, 42.4% and 34.5%, respectively. Our novel findings provided a better understanding of the epigenetic modulation of integrins and suggested that targeting the epigenetic up-regulated integrins to abrogate the migration activity might be a promising strategy to prevent HCC progression.
Lin
Kuen-Tyng
KT
Graduate Institute of Life Sciences, National Defense Medical Center, National Defense University, Taipei, Taiwan.
Yeh
Shiou-Hwei
SH
Chen
Ding-Shinn
DS
Chen
Pei-Jer
PJ
Jou
Yuh-Shan
YS
D001483:Base Sequence; D015153:Blotting, Western; D018821:CD18 Antigens / Q000235:genetics* / Q000502:physiology; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology*; D017931:DNA Primers; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D044127:Epigenesis, Genetic; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology*; D039441:Integrin alpha4 / Q000235:genetics* / Q000502:physiology; D039641:Integrin beta Chains / Q000235:genetics* / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation / Q000187:drug effects
D018821:CD18 Antigens; D017931:DNA Primers; D004791:Enzyme Inhibitors; D006877:Hydroxamic Acids; D039641:Integrin beta Chains; C072170:integrin beta6; D039441:Integrin alpha4; C012589:trichostatin A