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中国生物工程杂志

China Biotechnology
China Biotechnology  2017, Vol. 37 Issue (3): 18-26    DOI: 10.13523/j.cb.20170303
    
Construction of the Stable Prostate Cancer Cell Lines Overexpress GPRC6A and EMT Study
SHENG Bin1, YANG Fan2, SUN Xin-yi1, CHEN Yao1, LI Li1, YANG Jian-yi1, DU Sheng-jia1, LIU Ming1
1. School of Basic Medical Science, Shanxi Medical University, Taiyuan 030001, China;
2. Peking Union Medical College and Chinese Academy of Medical Sciences, Graduate School, Beijing 100005, China
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Abstract  

Objective:To construct the prostate cancer cell line LncapC4-2 that overexpress GPRC6A gene, detect the cell migration and invasion abilities and the expression of EMT related genes, thus to establish the basis of exploring the role of EMT induced by GPRC6A in prostate cancer progression. Methods:The GPRC6A overexpression lentivirus vector pCDH-GPRC6A was constructed, The lentivirus were packaged using human embryonic kidney 293T cells and LncapC4-2 cell line was infected, and GPRC6A stably overexpressing cell line was screened using puromycin. RT-qPCR and Western blot methods were used to verify the overexpression of GPRC6A in the cell line, scratches and transwell cell migration assay were used to test the cell migration and invasion abilities and QPCR was used to detect the expressions of EMT-associated genes. Results:The lentivirus vector pCDH-GPRC6A was constructed successfully. In GPRC6A overexpression Lncap C4-2 cells, the GPRC6A expression was increased in mRNA and protein levels detected by using RT-PCR and Western blot methods. The cell migration and invasion abilities were higher in GPRC6A overexpression Lncap C4-2 cells than those in control cells. The expressions of EMT-associated gene E-cadherin and SNAIL were reduced and increased, respectively, in GPRC6A overexpression LncapC4-2 cells. Conclusion:The prostate cancer cell line LncapC4-2 that stably overexpress GPRC6A has been constructed successfully. GPRC6A maybe involves in the progression of prostate cancer by enhancing the abilities of cell migration and invasion, and promoting EMT in cancer cells.



Key wordsGPRC6A      EMT      Prostate cancer     
Received: 29 September 2016      Published: 25 March 2017
ZTFLH:  Q813  
Cite this article:

SHENG Bin, YANG Fan, SUN Xin-yi, CHEN Yao, LI Li, YANG Jian-yi, DU Sheng-jia, LIU Ming. Construction of the Stable Prostate Cancer Cell Lines Overexpress GPRC6A and EMT Study. China Biotechnology, 2017, 37(3): 18-26.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20170303     OR     https://manu60.magtech.com.cn/biotech/Y2017/V37/I3/18

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