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. 2017 Aug;14(2):1875-1883.
doi: 10.3892/ol.2017.6361. Epub 2017 Jun 9.

Wip1 gene silencing enhances the chemosensitivity of human colon cancer cells

Zhong-Sheng Xia  1 , Di Wu  1 , Wa Zhong  1 , Xi-Ji Lu  1 , Tao Yu  1 , Qi-Kui Chen  1
Affiliations

Wip1 gene silencing enhances the chemosensitivity of human colon cancer cells

Zhong-Sheng Xia et al. Oncol Lett. 2017 Aug.

Abstract

Colon cancer is one of the most common cancers in the world. Multidrug resistance is one of the main reasons for failure of therapy in patients with advanced colon cancer. In previous studies, multiple methods were investigated to reverse the multidrug resistance of colon cancer cells. However, to date, no clinical method has been identified to be satisfactory. Therefore, successful reversal of drug resistance in colon cancer cells still requires new therapeutic strategies or pharmaceuticals. Wild-type p53-induced phosphatase (Wip1), a member of the 2C type serine/threonine protein phosphatase family, is closely associated with the p53 gene, which is the most important tumor-suppressor gene. Wip1 was reported to be associated with the chemosensitivity of breast cancer cells. However, the correlation between the expression of Wip1 gene and the chemosensitivity of colon cancer cells has not been reported yet. In the present study, Wip1-811 small interfering RNA (siRNA) targeting Wip1 was investigated to reverse the multidrug resistance of colon cancer cells. The siRNA duplexes were transfected into RKO colon cancer cells. The messenger RNA (mRNA) expression of Wip1 was measured by reverse transcription-quantitative polymerase chain reaction. The protein level of Wip1 was detected by western blotting. The cell viability was measured by MTS assay. The cell apoptosis and cell cycle were analyzed by flow cytometry. Intracellular adriamycin cumulative concentration was determined using flow cytometry. Wip1-811 siRNA efficiently inhibited the expression of Wip1 at the mRNA and protein levels, and enhanced the sensitivity of RKO colon cancer cells towards chemotherapy, which was accompanied by increased cell apoptosis, following the inhibition of Wip1 gene expression. These results indicate that Wip1 gene silencing could enhance the chemosensitivity of colon cancer cells, which may provide a new potential approach for the reversal of multidrug resistance in colon cancer cells.

Keywords: Wip1 gene; chemosensitivity; colon cancer; small interfering RNA.

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Figures

Figure 1.
Figure 1.
Influence of different concentrations of Wip1-811 siRNA on the relative mRNA expression of Wip1 in RKO colon cancer cells (A) 24 and (B) 48 h after transfection. *P<0.05 vs. negative control. Wip1, wild-type p53-induced phosphatase; siRNA, small interfering RNA; mRNA, messenger RNA.
Figure 1.
Figure 1.
Influence of different concentrations of Wip1-811 siRNA on the relative mRNA expression of Wip1 in RKO colon cancer cells (A) 24 and (B) 48 h after transfection. *P<0.05 vs. negative control. Wip1, wild-type p53-induced phosphatase; siRNA, small interfering RNA; mRNA, messenger RNA.
Figure 2.
Figure 2.
Influence of different concentrations of Wip1-811 siRNA on the protein expression of Wip1 in RKO colon cancer cells (A) 24 and (B) 48 h after transfection. *P<0.05 vs. negative control. Wip1, wild-type p53-induced phosphatase; siRNA, small interfering RNA.
Figure 2.
Figure 2.
Influence of different concentrations of Wip1-811 siRNA on the protein expression of Wip1 in RKO colon cancer cells (A) 24 and (B) 48 h after transfection. *P<0.05 vs. negative control. Wip1, wild-type p53-induced phosphatase; siRNA, small interfering RNA.
Figure 3.
Figure 3.
Influence of Wip1-811 siRNA on the relative mRNA expression of Wip1 in RKO colon cancer cells after transfection for 24–96 h. *P<0.05 vs. negative control at the same time point. Wip1, wild-type p53-induced phosphatase; siRNA, small interfering RNA; mRNA, messenger RNA.
Figure 4.
Figure 4.
Influence of Wip1-811 siRNA on the protein expression of Wip1 in RKO colon cancer cells after transfection for 24–96 h. *P<0.05 vs. negative control at the same time point. Wip1, wild-type p53-induced phosphatase; siRNA, small interfering RNA.
Figure 5.
Figure 5.
Influence of Wip1-811 siRNA on cell apoptosis in RKO colon cancer cells treated with 5-FU. A, Control; B, Control+5-FU; C, Negative control+5-FU; D, Wip1-811 siRNA; E, Liposome+5-FU; F, Wip1-811 siRNA+5-FU. ∆P>0.05 vs. control; #P<0.05 vs. Wip1-811 siRNA group; *P<0.05 vs. negative control. Wip1, wild-type p53-induced phosphatase; siRNA, small interfering RNA; 5-FU, 5-fluorouracil; Annexin V-FITC, Annexin V-fluorescein isothiocyanate; PI, Propidium iodide; UL, upper left; UR, upper right; LL, lower left; LR, lower right.
Figure 6.
Figure 6.
Influence of Wip1-811 siRNA on cell apoptosis in RKO colon cancer cells treated with oxaliplatin. A, Control; B, Control+oxaliplatin; C, Negative control+oxaliplatin; D, Wip1-811 siRNA; E, Liposome+5oxaliplatin; F, Wip1-811 siRNA+oxaliplatin. ∆P>0.05 vs. control; #P<0.05 vs. Wip1-811 siRNA group; *P<0.05 vs. negative control. Wip1, wild-type p53-induced phosphatase; siRNA, small interfering RNA; Annexin V-FITC, Annexin V-fluorescein isothiocyanate; PI, Propidium iodide; UL, upper left; UR, upper right; LL, lower left; LR, lower right.
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