Influence of ezrin-shRNA in combination with HSP70 on the apoptosis and proliferation of osteosarcoma cells

Yao,Qin; Pei,Yihua; Zhuo,Huiqin; Xie,Bozhen

doi:10.3892/ol.2016.5103

Influence of ezrin-shRNA in combination with HSP70 on the apoptosis and proliferation of osteosarcoma cells

Authors:
- Qin Yao
- Yihua Pei
- Huiqin Zhuo
- Bozhen Xie
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Affiliations: Central Laboratory, Zhongshan Hospital Xiamen University, Xiamen, Fujian 361004, P.R. China, Department of Spine Surgery, Zhongshan Hospital Xiamen University, Xiamen, Fujian 361004, P.R. China
Published online on: September 8, 2016 https://doi.org/10.3892/ol.2016.5103
Pages: 3441-3448
Copyright: © Yao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Ezrin and heat shock protein (HSP)70 have been reported to regulate cell apoptosis and tumor development of osteosarcoma. However, there has not been reported the synergy effect of knocking down ezrin and overexpressing HSP70. In the present study, two vectors, pGFP‑V‑RS‑shRNA and pGFP‑V‑RS‑shRNA‑HSP70, were constructed and transfected into LM8 cells [denoted as small hairpin (sh)RNA group and dual group, respectively]. The apoptosis rates in these two transfected groups were significantly higher than those in the control group (empty vector) (P=0.036), while significantly lower proliferation rates were observed in these two groups (P=0.023). The cytotoxic T lymphocyte activity on target LM8 tumor cells in the dual group was significantly higher than in other groups, with cytotoxicity as high as 55.56±2.10%. Further studies revealed that the transfection of ezrin‑shRNA/HSP70 also suppressed tumor formation in vivo in nude mice. A lower cluster of differentiation (CD)4/CD8 ratio was detected in the tumor formed by injecting cells in the dual group (P=0.006). Furthermore, the serum level of interleukin‑4 in the dual group was significantly decreased, while the serum level of interferon‑γ was significantly increased, compared with the other two groups (P=0.004). Simultaneously knocking down ezrin and overexpressing HSP70 promotes cellular apoptosis and suppresses the proliferation of osteosarcoma cells in vitro, and enhances the tumor killing effects of HSP70‑induced immune killing.

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