Intest Res.  2013 Jan;11(1):28-33. 10.5217/ir.2013.11.1.28.

The Effect of Transfection of RET9 and RET51 on Apoptosis in SW48 and RKO Colon Cancer Cells

Affiliations
  • 1Department of Internal Medicine, Ulsan University College of Medicine, Gangneung Asan Hospital, Gangneung, Korea.
  • 2Department of Internal Medicine, Sungkyunkwan University College of Medicine, Kangbuk Samsung Hospital, Seoul, Korea. diksmc.park@samsung.com

Abstract

BACKGROUND/AIMS
Dependent receptor can transmit both positive signal: proliferation, differentiation or migration; and negative signal: apoptosis. It depends on the presence of its ligand. This study was performed to determine the effects of transfection of dependent receptors in human colon cancer cell lines.
METHODS
Two dependent receptors (rearranged during transfection [RET]9 and RET51) were transfected into three human colon cancer cell lines: SW48, RKO and V400. Then, half of them were treated with glial cell line-derived neurotrophic factor (GDNF). Using ELISA and caspase assay, apoptosis was measured. Dose-response relation between GDNF and apoptosis was also analyzed. A pcDNA was used as an empty vector.
RESULTS
After transfection of RET51, apoptosis was increased in SW48 (70% with ELISA and 119% with caspase assay) and RKO (255% with ELISA and 106% with caspase assay) cell lines when compared with the pcDNA group. V400 cell line did not show increased apoptosis. Transfection of RET9 did not induce apoptosis in all of the three human colon cancer cell lines. Treatment with GDNF 12 hours after transfection of RET51 decreased apoptosis in SW48 (66% with ELISA and 60% with caspase assay) and RKO (39% with ELISA and 57% with caspase assay) when compared with the cell lines transfected with RET51 only. Apoptosis was down-regulated with increasing concentration of GDNF in RKO cell line.
CONCLUSIONS
This study showed that the apoptosis of human colon cancer cell line can be controlled by manipulating the dependent receptors and its ligands. We present the possibility of therapeutic method using dependent receptor in colon cancer.

Keyword

Colonic neoplasms; Transfection; Dependent receptor; Apoptosis, Ligand

MeSH Terms

Apoptosis
Cell Line
Colon
Colonic Neoplasms
Enzyme-Linked Immunosorbent Assay
Glial Cell Line-Derived Neurotrophic Factor
Humans
Ligands
Transfection
Glial Cell Line-Derived Neurotrophic Factor
Ligands
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