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G-protein coupled receptor 34 knockdown impairs the proliferation and migration of HGC-27 gastric cancer cells in vitro.

Jin ZT, Li K, Li M, Ren ZG, Wang FS, Zhu JY, Leng XS, Yu WD - Chin. Med. J. (2015)

Bottom Line: We also measured expression profile of PI3K/PDK1/AKT and ERK using Western blotting.The ShRNA directed against GPR34 effectively inhibited both endogenous mRNA and protein expression levels of GPR34, and significantly down-regulated the expression of PIK3CB (P < 0.01), PIK3CD (P < 0.01), PDK1 (P < 0.01), phosphorylation of PDK1 (P < 0.01), Akt (P < 0.01), and ERK (P < 0.01).Furthermore, GPR34 knockdown resulted in an obvious reduction in HGC-27 cancer cell proliferation and migration activity (P < 0.01).

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology; Institute of Clinical Molecular Biology, Peking University People's Hospital, Beijing 100044, China.

ABSTRACT

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. The current study aimed to investigate the effect of GPR34 knockdown on the proliferation, migration, and apoptosis of HGC-27 gastric cancer cells and the underlying mechanisms.

Methods: The expression of GPR34 in gastric cancer cell line HGC-27 was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. HGC-27 cells were employed to construct the stable GPR34 knockdown cell model in this study. Real-time RT-PCR and Western blotting were applied to validate the effect of short hairpin RNA (ShRNA) on the expression of GPR34 in HGC-27 gastric cells. The proliferation, migration of these cells were examined by Cell Counting Kit-8 and transwell. We also measured expression profile of PI3K/PDK1/AKT and ERK using Western blotting.

Results: The ShRNA directed against GPR34 effectively inhibited both endogenous mRNA and protein expression levels of GPR34, and significantly down-regulated the expression of PIK3CB (P < 0.01), PIK3CD (P < 0.01), PDK1 (P < 0.01), phosphorylation of PDK1 (P < 0.01), Akt (P < 0.01), and ERK (P < 0.01). Furthermore, GPR34 knockdown resulted in an obvious reduction in HGC-27 cancer cell proliferation and migration activity (P < 0.01).

Conclusions: GPR34 knockdown impairs the proliferation and migration of HGC-27 gastric cancer cells in vitro and provides a potential implication for therapy of gastric cancer.

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Related in: MedlinePlus

GPR34 expression involved in gastric cell proliferation and migration (HGC-27 cell) in vitro by ERK and PIK3/AKT pathways. (a) Basal expression of p110 (α, β, and δ) as compared to HGC-27 and HGC-27-V. Only p110β was found to be significantly downregulated in HGC-27-K (*P< 0.01). The phosphorylated PDK1, AKT, and ERK were constitutively suppressed; (b) Densitometry analysis of all tested proteins in Western blotting was performed using ImageJ software (*P< 0.01).
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Figure 5: GPR34 expression involved in gastric cell proliferation and migration (HGC-27 cell) in vitro by ERK and PIK3/AKT pathways. (a) Basal expression of p110 (α, β, and δ) as compared to HGC-27 and HGC-27-V. Only p110β was found to be significantly downregulated in HGC-27-K (*P< 0.01). The phosphorylated PDK1, AKT, and ERK were constitutively suppressed; (b) Densitometry analysis of all tested proteins in Western blotting was performed using ImageJ software (*P< 0.01).

Mentions: Combined with our previous study,[14] to understand decreased expression of GPR34 in HGC-27 represents a potential molecular mechanism by which HGC-27-K results in the decreased abilities of proliferation and migration any differently from that seen with HGC-27 parental cells in vitro. We studied alterations in ERK, the class I PI3K subunits (p110α, p110β, and p110δ), and PDK1/AKT. When compared with HGC-27 and HGC-27-V cells, both p110β and p110δ were found to be down-regulated in HGC-27-K cells. We did not observe any alteration in the expression of p110α and detect the expression of p110γ in either cell-line [Figure 5]. Furthermore, low expression levels of p-PDK1, p-AKT, and p-ERK were observed in HGC-27-K cells [Figure 5].


G-protein coupled receptor 34 knockdown impairs the proliferation and migration of HGC-27 gastric cancer cells in vitro.

Jin ZT, Li K, Li M, Ren ZG, Wang FS, Zhu JY, Leng XS, Yu WD - Chin. Med. J. (2015)

GPR34 expression involved in gastric cell proliferation and migration (HGC-27 cell) in vitro by ERK and PIK3/AKT pathways. (a) Basal expression of p110 (α, β, and δ) as compared to HGC-27 and HGC-27-V. Only p110β was found to be significantly downregulated in HGC-27-K (*P< 0.01). The phosphorylated PDK1, AKT, and ERK were constitutively suppressed; (b) Densitometry analysis of all tested proteins in Western blotting was performed using ImageJ software (*P< 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836262&req=5

Figure 5: GPR34 expression involved in gastric cell proliferation and migration (HGC-27 cell) in vitro by ERK and PIK3/AKT pathways. (a) Basal expression of p110 (α, β, and δ) as compared to HGC-27 and HGC-27-V. Only p110β was found to be significantly downregulated in HGC-27-K (*P< 0.01). The phosphorylated PDK1, AKT, and ERK were constitutively suppressed; (b) Densitometry analysis of all tested proteins in Western blotting was performed using ImageJ software (*P< 0.01).
Mentions: Combined with our previous study,[14] to understand decreased expression of GPR34 in HGC-27 represents a potential molecular mechanism by which HGC-27-K results in the decreased abilities of proliferation and migration any differently from that seen with HGC-27 parental cells in vitro. We studied alterations in ERK, the class I PI3K subunits (p110α, p110β, and p110δ), and PDK1/AKT. When compared with HGC-27 and HGC-27-V cells, both p110β and p110δ were found to be down-regulated in HGC-27-K cells. We did not observe any alteration in the expression of p110α and detect the expression of p110γ in either cell-line [Figure 5]. Furthermore, low expression levels of p-PDK1, p-AKT, and p-ERK were observed in HGC-27-K cells [Figure 5].

Bottom Line: We also measured expression profile of PI3K/PDK1/AKT and ERK using Western blotting.The ShRNA directed against GPR34 effectively inhibited both endogenous mRNA and protein expression levels of GPR34, and significantly down-regulated the expression of PIK3CB (P < 0.01), PIK3CD (P < 0.01), PDK1 (P < 0.01), phosphorylation of PDK1 (P < 0.01), Akt (P < 0.01), and ERK (P < 0.01).Furthermore, GPR34 knockdown resulted in an obvious reduction in HGC-27 cancer cell proliferation and migration activity (P < 0.01).

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology; Institute of Clinical Molecular Biology, Peking University People's Hospital, Beijing 100044, China.

ABSTRACT

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. The current study aimed to investigate the effect of GPR34 knockdown on the proliferation, migration, and apoptosis of HGC-27 gastric cancer cells and the underlying mechanisms.

Methods: The expression of GPR34 in gastric cancer cell line HGC-27 was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. HGC-27 cells were employed to construct the stable GPR34 knockdown cell model in this study. Real-time RT-PCR and Western blotting were applied to validate the effect of short hairpin RNA (ShRNA) on the expression of GPR34 in HGC-27 gastric cells. The proliferation, migration of these cells were examined by Cell Counting Kit-8 and transwell. We also measured expression profile of PI3K/PDK1/AKT and ERK using Western blotting.

Results: The ShRNA directed against GPR34 effectively inhibited both endogenous mRNA and protein expression levels of GPR34, and significantly down-regulated the expression of PIK3CB (P < 0.01), PIK3CD (P < 0.01), PDK1 (P < 0.01), phosphorylation of PDK1 (P < 0.01), Akt (P < 0.01), and ERK (P < 0.01). Furthermore, GPR34 knockdown resulted in an obvious reduction in HGC-27 cancer cell proliferation and migration activity (P < 0.01).

Conclusions: GPR34 knockdown impairs the proliferation and migration of HGC-27 gastric cancer cells in vitro and provides a potential implication for therapy of gastric cancer.

Show MeSH
Related in: MedlinePlus