Background: Overexpression of G-protein coupled receptor 34 (GPR34) affects the progression and prognosis of human gastric adenocarcinoma, however, the role of GPR34 in gastric cancer development and progression has not been well-determined.
G protein - coupled receptor 34 (GPR34) is a 7-transmembrane receptor.
In this study, we investigated the effects of down-regulation of GPR34 by ShRNA on proliferation, apoptosis, and migration of HGC-27 gastric cancer cells, as well as PI3K/AKT and ERK expression, with the aim of evaluating whether the expression of GPR34 may be linked to gastric cancer progression, and exploring the underlying mechanisms.
One MSCV-based retroviral vector containing-shRNA that specifically target GPR34 transcript was purchased from Open Biosystems (Huntsville, AL, USA).
Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) was performed using an OPTICON 2 real-time PCR detection system (Bio-Rad, USA) using human GPR34 primers [sup] combined with SYBR Green Master Mix (Applied Biosystems, USA).
(GADPH primers: Forward, 5'- CTGGTAAAGT GGATATTGTTGCCAT-3', reverse, 5'-TGGAATCATA TTGGAACATGTAAACC-3'; human GPR34 primers (forward: 5'- CTCCCACAGAATGCGCTTTATA-3', reverse: 5'-CAACCAGTCCCACGATGAAAA-3').
Construction of HGC-27 GPR34 knock-down cell models
HGC-27, a low-phosphatase and tensin homolog gastric cancer cell line, was selected and used as a cell model in vitro to determine the correlation between GPR34 expression and proliferation, apoptosis and migration of gastric cancer cells.
GPR34 knockdown impairs the proliferation of HGC-27 gastric cancer cells
Significant up-regulation of GPR34 protein in gastric cancer tissues suggested a potential oncogenic role of this gene.
GPR34 knockdown dose not induce apoptosis of HGC-27 gastric cancer cells