Intron‑specific shRNA‑mediated downregulation of survivin and promotion of apoptosis in HeLa cells

Wang,Yue‑Li; Shao,Xin; Wang,Fa; Zeng,Liang; Hu,Liang; Cui,Shi‑Quan; Hou,Gan; Huang,Di‑Nan

doi:10.3892/ol.2017.6996

Intron‑specific shRNA‑mediated downregulation of survivin and promotion of apoptosis in HeLa cells

Authors:
- Yue‑Li Wang
- Xin Shao
- Fa Wang
- Liang Zeng
- Liang Hu
- Shi‑Quan Cui
- Gan Hou
- Di‑Nan Huang
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Affiliations: Department of Basic Medicine, Guangdong Medical University, Zhanjiang, Guangdong 524023, P.R. China, People's Hospital of Shiyan, Shiyan, Hubei 442000, P.R. China, Department of Clinical Biochemistry, Guangdong Medical University, Dongguan, Guangdong 523808, P.R. China
Published online on: September 18, 2017 https://doi.org/10.3892/ol.2017.6996
Pages: 5927-5933
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Overexpression of the survivin gene contributes to tumorigenesis; it has been recognized as an important target for cancer therapy. In the present study, survivin expression was suppressed using recombinant plasmid mediated short hairpin RNAs (shRNAs) that were constructed to target exonic or intronic sequences of the survivin gene. In addition, a negative control shRNA was constructed. HeLa cells were transfected with specific shRNA constructs and the blocking efficiency of each shRNA was assessed at the mRNA and protein levels; and the five shRNA constructs with higher blocking efficiency were selected. Cell apoptosis was assessed by flow cytometry (FCM) following Annexin V‑fluorescein isothiocyanate/propidium iodide double staining. Hoechst staining was used to detect the morphological diversity of the nuclei in apoptotic cells. The results demonstrated that survivin expression was effectively reduced by the transfection of shRNAs in HeLa cells. In addition, the apoptotic rates of the shRNA‑treated groups were significantly increased compared with the negative control group according to the FCM results. The nuclei of HeLa cells exhibited apoptotic characteristics in the shRNA‑treated groups as identified by Hoechst staining. Survivin‑targeting shRNAs effectively downregulated the expression of the gene and markedly increased the apoptotic rate of HeLa cells. Data from the present study also indicated that the intron‑specific shRNA demonstrate a high efficiency of inhibition of survivin expression and were able to induce cell apoptosis of HeLa cells through RNAi, potentially providing novel target sites for tumor therapy. In conclusion, the present study suggests that intron‑specific blocking of survivin by RNAi may provide a tool for anticancer therapy.

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