Ann Surg Treat Res.  2016 Dec;91(6):273-277. 10.4174/astr.2016.91.6.273.

Silencing the livin gene enhances the cytotoxic effects of anticancer drugs on colon cancer cells

• Oh BY
• Kim KH
• Chung SS
• Lee RA

• KMID: 2360386
• DOI: http://doi.org/10.4174/astr.2016.91.6.273

Abstract

PURPOSE
Livin is associated with drug response in several cancers. The aim of this study was to investigate the effect of silencing the livin gene expression on anticancer drug response in colorectal cancer.
METHODS
siRNA was transfected at different concentrations (0, 10, and 30nM) into HCT116 cells, then cells were treated with either 5-fluorouracil (FU)/leucovorin (LV) or oxaliplatin (L-OHP)/5-FU/LV. Cellular viability and apoptosis were evaluated following silencing of livin gene expression combined with treatment with anticancer drugs.
RESULTS
Livin gene expression was effectively suppressed by 30nM siRNA compared with control and 10nM siRNA. The 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay showed that proliferation was effectively inhibited in cells treated with a combination of both siRNA and an anticancer drug, compared to cells treated with siRNA-Livin or anticancer drug alone. In particular, the combination of 30nM siRNA and L-OHP/5-FU/LV resulted in a 93.8% and 91.4% decrease, compared to untreated control or L-OHP/5-FU/LV alone, respectively. Cellular proliferation was most effectively suppressed by a combination of 30nM of siRNA and L-OHP/5-FU/LV compared to other combinations.
CONCLUSION
siRNA-mediated down-regulation of livin gene expression could significantly suppress colon cancer growth and enhance the cytotoxic effects of anticancer drugs such as 5-FU and L-OHP. The results of this study suggest that silencing livin gene expression in combination with treatment with anticancer drugs might be a novel cancer therapy for colorectal cancer.

Keyword

Livin protein; Oxaliplatin; Apoptosis; Colorectal neoplasms

MeSH Terms

Apoptosis
Cell Proliferation
Colon*
Colonic Neoplasms*
Colorectal Neoplasms
Down-Regulation
Fluorouracil
Gene Expression
HCT116 Cells
RNA, Small Interfering
Fluorouracil
RNA, Small Interfering

Figure

• 

  Fig. 1 Silencing of livin gene expression after siRNA transfection. Livin gene expression was effectively suppressed after transfection of 30nM siRNA in HCT116 cells. Livin mRNA detected by reverse transcription-polymerase chain reaction. Columns = means ± standard deviation (n = 3). *P < 0.05. **P < 0.01. ***P < 0.001.

• 

  Fig. 2 Antiproliferative effect of silencing livin gene in combination with anticancer drug treatment. Proliferation of cells treated with combination of siRNA-Livin and anticancer drug was effectively inhibited. This effect was most prominent with combination of 30nM siRNA and oxaliplatin/5-fluorouracil/leucovorin. Cell viability was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Columns = means ± standard deviation (n = 3). *P < 0.05. **P < 0.01. ***P < 0.001.

• 

  Fig. 3 Caspase activity after siRNA transfection and treatment with anticancer drugs. Expression of caspase 3 and 7 showed greater decrease in cells treated with combination of 30nM siRNA and oxaliplatin/5-fluorouracil/leucovorin (L-OHP/5-FU/LV) compared with either siRNA or L-OHP/5-FU/LV alone. Expression of caspase 3 and caspase 7 was evaluated by reverse transcription-polymerase chain reaction.

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