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.2018 Oct;70(5):1315-1323.
doi: 10.1007/s10616-018-0221-9. Epub 2018 May 25.

Knockdown of ribosomal protein S15A inhibits proliferation of breast cancer cells through induction of apoptosis in vitro

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Knockdown of ribosomal protein S15A inhibits proliferation of breast cancer cells through induction of apoptosis in vitro

Weiliang Feng et al. Cytotechnology.2018 Oct.

Abstract

To explore the role of ribosomal protein S15A (RPS15A) in breast cancer. The Oncomine database was used to compare the expression of RPS15A in human breast cancer tissues and normal tissues. RPS15A in breast cancer cell line ZR-75-30 and BT474 was specifically knocked down using lentivirus-mediated short hairpin RNAs (shRNAs). RPS15A knockdown efficiency was validated by quantitative polymerase chain reaction and western blot analysis. Subsequently, the functional effects of RPS15A on proliferation of breast cancer cells were investigated by MTT, colony formation and flow cytometry assays. Functional analysis indicated that RPS15A knockdown could inhibit cell proliferation, induced cell cycle arrest and apoptosis. Mechanism analysis revealed RPS15A mediated apoptosis via activating of caspase-3 and PARP cleavage, upregulating of Bad and BAX and downregulating of Bcl-2. Our preliminary study highlighted the importance of RPS15A in breast cancer growth. The inhibition of RPS15A may be a promising therapeutic target for breast cancer treatment.

Keywords: Apoptosis; Breast cancer; Cell proliferation; PRS15A.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Knockdown efficiency of RPS15A by lentivirus transduction of the breast cancer cells, ZR-75-30 and BT474.a Representative microscopic images of breast cancer cells were shown. Transduction efficiency was evidences by the expression of the visible green fluorescence protein (GFP).b RT-PCR was used to analyze the RPS15A mRNA level in three groups of ZR-75-30 and BT474 cells after 5 days infection.c RPS15A Protein levels were detected by western blot analysis in ZR-75-30 and BT474 cells 4 days after transduction. ***P < 0.001; Scale bars, 10 μm
Fig. 2
Fig. 2
RPS15A knockdown inhibited cell proliferation and colony formation in ZR-75-30 and BT474 cells.a Cell proliferation was significantly inhibited after RPS15A knockdown, which was examined by the MTT assay.b Representative photographs of colonies showed a significant inhibition of colony formation in the shRPS15A (S1) condition.c Statistical analysis of the number of colonies (>= 50 single cells) stained with crystal violet. ***P < 0.001; Scale bars, 25 μm
Fig. 3
Fig. 3
Knockdown of RPS15A induces cell-cycle arrest.a Cell cycle distribution of ZR-75-30 cells was analyzed by flow cytometry.b,c Statistical analysis of the percentage of cells in the different cell cycle phases **P < 0.01; ***P < 0.001, Scale bars, 10 μm
Fig. 4
Fig. 4
Knockdown of RPS15A promotes apoptosis in ZR-75-30 cells.a Representative images showing Annexin V/7-ADD staining results in ZR-75-30 cells.b Representative images showing Annexin V/7-ADD staining results in BT474 cells.c,d Quantification ofa andb.e,f Expression of downstream proteins related to apoptosis in ZR-75-30 and BT474 cells by western blot analysis. ***P < 0.001; Scale bars, 10 μm
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