The effect of NaB, SAHA and TSA on expression of the tumor suppressor genes SPINT2 and NDRG2 in LoVo cells

Sascha Gyrud, Liv Nordahl Terkelsen, Malte Wegan Frier, Anne-Line Strange Laursen, Sif Baungaard & Emilie Kyhse Bender

Studenteropgave: Fagmodulprojekt

Abstrakt

Colon cancer is a disease affecting an increasing number of people and therefore new
treatments are in constant development. Research suggests that drugs which utilize epigenetic
mechanisms can possibly be used for treatment. One of these epigenetic mechanisms is
histone acetylation which alter the accessibility of the genome for the transcriptional
machinery. Histone deacetylases (HDACs) remove acetyl groups on histones of the genome.
The drugs under investigation utilize this mechanism by being inhibitors of HDACs
(HDACis) and is speculated to have an anti-tumor effect by upregulating specific tumor
suppressor genes as they generally increase transcription. The purpose of this project is to
examine how the expression of the tumor suppressor genes Serine protease inhibitor 2
( SPINT2 ) and N-myc downstream-regulated gene 2 ( NDRG2 ) in a colon cancer cell line,
LoVo, is affected by treatment with three HDACis: Sodium Butyrate (NaB), Suberoylanilide
Hydroxamic Acid (SAHA) and Trichostatin A (TSA). Our hypothesis is that inhibition of
HDAC activity can upregulate SPINT2 and NDRG2 expression through histone acetylation
and thereby exhibit anti-tumor effects. In order to examine the expression of the tumor
suppressor genes, histone acetylation and morphological changes, the methods RNA
extraction, cDNA synthesis, PCR, gel electrophoresis; protein extraction, protein gel
electrophoresis, Western blot; and phase-contrast microscopy were used, respectively.
Furthermore, the results from the experiments are discussed and brought in relation to current
literature and other experiments conducted investigating the same aspects.
Based on the results, it is not possible to determine how the expression of SPINT2 and
NDRG2 is affected by HDACis, but other studies support our hypothesis. In addition, induced
apoptosis and corresponding increased acetylation of histones might indicate an upregulation
of the tumor suppressor genes by HDACi treatment.

UddannelserMolekylærbiologi, (Bachelor/kandidatuddannelse) Bachelor
SprogEngelsk
Udgivelsesdato28 maj 2018
Antal sider97

Emneord

  • Epigenetics
  • HDACi
  • SAHA
  • NaB
  • TSA
  • SPINT2
  • NDRG2
  • Colorectal cancer
  • LoVo cells
  • tumor suppressor genes
  • histone acetylation