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. 2018 Oct;16(4):4279-4284.
doi: 10.3892/ol.2018.9172. Epub 2018 Jul 18.

Effects of RSF-1 on proliferation and apoptosis of breast cancer cells

Yuhui Liu  1 , Junda Gai  1 , Lin Fu  1 , Xiuwei Zhang  2 , Enhua Wang  1 , Qingchang Li  1
Affiliations

Effects of RSF-1 on proliferation and apoptosis of breast cancer cells

Yuhui Liu et al. Oncol Lett. 2018 Oct.

Abstract

Effect of interference with chromatin remodeling and spacing factor-1 (RSF-1) on proliferation and apoptosis of breast cancer cells was investigated. MCF-7 and SKBR-3 cells were cultured in vitro and were divided into 3 groups: control group, negative siRNA control group (NC) and RSF-1 siRNA group. Western blot analysis was used to detect the expression of RSF protein after interference. Cell Counting Kit-8 (CCK-8) method was used to detect the effect of RSF-1 siRNA on cell proliferation. Plate cloning assay was used to detect the effect of RSF-1 siRNA on cell clone formation ability. Annexin V/PI double staining method was used to detect the effect of RSF-1 siRNA on cell apoptosis. Effect of RSF-1 siRNA on nuclear factor-κB (NF-κB) and its downstream signaling pathway were detected by western blot analysis. Western blot analysis showed that RSF-1 siRNA significantly downregulated the expression of RSF-1 protein in MCF-7 and SKBR-3 cells at 72 h after transfection (P<0.01). Cell proliferation assay showed that RSF-1 siRNA significantly reduced the proliferation ability and clone formation ability of MCF-7 and SKBR-3 cells compared with the control group (P<0.01). Annexin V/PI double staining assay results showed that compared with the control group, RSF-1 siRNA significantly increased the apoptosis rate of MCF-7 and SKBR-3 cells (P<0.01). Helenalin and Rsf-1 siRNA significantly reduced the expression levels of p-p65, Bcl-2, and XIAP proteins (P<0.01). Interfering with the expression of RSF-1, gene can effectively inhibit the proliferation of MCF-7 and SKBR-3 cells and promote their apoptosis. RSF-1 can be used as a potential new therapeutic target for the treatment of breast cancer.

Keywords: MCF-7; NF-κB; RSF-1; SKBR-3; apoptosis; proliferation.

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Figures

Figure 1.
Figure 1.
Western blot analysis of effect of RSF-1 siRNA interference on RSF-1 protein expression. (A) Expression of RSF-1 protein in MCF-7 cells. (B) Expression of RSF-1 protein in SKBR-3 cells. **P<0.01, compared with control group; ##P<0.01, compared with negative control group.
Figure 2.
Figure 2.
Cell proliferation detected by CCK-8. (A) Proliferation rate of MCF-7 cells. (B) Proliferation rate of SKBR-3 cells. **P<0.01, compared with control group; ##P<0.01, compared with negative control group.
Figure 3.
Figure 3.
Cell clone formation assay results. (A) Representative results of MCF-7 and SKBR-3 cell clone formation. (B) Normalized MCF-7 cell clone formation data. (C) Normalized SKBR-3 cell clone formation data. **P<0.01, compared with control group; ##P<0.01, compared with negative control group.
Figure 4.
Figure 4.
Annexin V/PI staining to detect the effect of RSF-1 siRNA on cell apoptosis. (A) Representative results of MCF-7 cells; (B) normalized data of MCF-7 cells; (C) representative results of SKBR-3 cells; (D) normalized data of SKBR-3 cells. **P<0.01, compared with control group; ##P<0.01, compared with negative control group.
Figure 5.
Figure 5.
Western blot analysis of the effect of RSF-1 siRNA on p65 and its downstream signaling pathways. (A) Representative results of MCF-7 cells; (B) normalized data of MCF-7 cells; (C) representative results of SKBR-3 cells; (D) normalized data of SKBR-3 cells.
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