Targeting of NHERF1 through RNA interference inhibits the proliferation and migration of metastatic prostate cancer cells
Targeting of NHERF1 through RNA interference inhibits the proliferation and migration of metastatic prostate cancer cells
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通过RNA干扰靶向NHERF1抑制转移性前列腺癌细胞的增殖和迁移
DOI：
10.3892/ol.2015.4007
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发表时间：
2016-02-01
期刊：
影响因子：
2.9
通讯作者：
Song, Naling
中科院分区：
文献类型：
--
作者：
Ma, Qiang;Jiao, Yanna;Song, Naling
本研究旨在探讨利用短发夹RNA（short-hairpin RNA，shRNA）技术下调Na+/H+交换调节因子1（Na+/H+ exchanger regulatory factor 1，NHERF 1）基因对前列腺癌细胞恶性行为的影响。超级。使用Lipofectamine 2000将puro NHERF 1 shRNA载体转染到PC-3 M前列腺癌细胞中。获得稳定的细胞系，并通过蛋白质印迹分析验证NHERF 1敲低。然后使用MTT测定来测量PC-3 M细胞增殖;此外，使用伤口愈合测定来评估细胞迁移。采用流式细胞术以确定NHERF 1敲低对细胞凋亡的影响。然后通过蛋白质印迹分析确定凋亡途径蛋白B细胞淋巴瘤-2（Bcl-2）和Bcl-2相关X蛋白的表达水平。结果表明，NHERF 1的shRNA敲低显著抑制PC-3 M细胞的增殖，抑制率> 50%。此外，敲低NHERF 1显著抑制PC-3 M细胞的迁移。携带NHERF 1 shRNA的PC-3 M细胞表现出显著增加的凋亡，与亲本PC-3 M细胞和用空载体转染的细胞相比，增加了接近4倍。此外，结果显示，敲低NHERF 1降低Bcl-2的蛋白表达，但Bax的表达没有改变。总之，使用shRNA敲低NHERF 1抑制PC-3 M细胞的增殖和迁移并促进凋亡，突出了NHERF 1在前列腺癌进展中的作用。
The present study aimed to investigate the effects of Na+/H+ exchanger regulatory factor1 (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 similar to 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.