Targeting of NHERF1 through RNA interference inhibits the proliferation and migration of metastatic prostate cancer cells

Ma,Qiang; Jiao,Yanna; Hao,Yumei; Yan,Shanshan; Lyu,Nenan; Gao,Honglin; Li,Deguan; Liu,Qiang; Zheng,Junfang; Song,Naling

doi:10.3892/ol.2015.4007

Targeting of NHERF1 through RNA interference inhibits the proliferation and migration of metastatic prostate cancer cells

Authors:
- Qiang Ma
- Yanna Jiao
- Yumei Hao
- Shanshan Yan
- Nenan Lyu
- Honglin Gao
- Deguan Li
- Qiang Liu
- Junfang Zheng
- Naling Song
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Affiliations: Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Institute of Radiation Medicine, Peking Union Medical College and Chinese Academy of Medical Science, Tianjin 300192, P.R. China, Department of Genetics, National Research Institute for Family Planning, Beijing 100081, P.R. China, Department of Molecular Biology, Institute of Medical Biology, Peking Union Medical College and Chinese Academy of Medical Science, Kunming, Yunnan 650000, P.R. China, Department of Gynecologic Oncology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, P.R. China, Department of Biochemistry and Molecular Biology, Capital Medical University, Beijing 100069, P.R. China
Published online on: December 4, 2015 https://doi.org/10.3892/ol.2015.4007
Pages: 1149-1154

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Abstract

The present study aimed to investigate the effects of Na+/H+ exchanger regulatory factor 1 (NHERF1) gene knockdown, using short‑hairpin RNA (shRNA), on the malignant behaviors of prostate cancer cells. A pSuper.puro NHERF1 shRNA vector was transfected into PC‑3M prostate cancer cells using Lipofectamine 2000. Stable cell lines were obtained and NHERF1 knockdown was verified through western blot analysis. MTT assays were then used to measure PC‑3M cell proliferation; in addition, cell migration was assessed using a wound healing assay. Flow cytometry was employed in order to determine the effects of NHERF1 knockdown on apoptosis. Expression levels of apoptotic pathway proteins B cell lymphoma‑2 (Bcl‑2) and Bcl‑2‑associated X protein were then determined by western blot analysis. The results demonstrated that shRNA knockdown of NHERF1 significantly suppressed the proliferation of PC‑3M cells by >50%. In addition, knockdown of NHERF1 significantly inhibited the migration of PC‑3M cells. PC‑3M cells harboring NHERF1 shRNA exhibited significantly increased apoptosis, with an ~4‑fold increase compared with that of the parental PC‑3M cells and cells transfected with an empty vector. Furthermore, the results revealed that knockdown of NHERF1 reduced the protein expression of Bcl‑2, although the expression of Bax was unaltered. In conclusion, NHERF1 knockdown using shRNA inhibited the proliferation and migration of PC‑3M cells and promoted apoptosis, highlighting the role of NHERF1 in prostate cancer progression.

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