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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

The effect of GPR34 on migration of HGC-27 cells In vitro. Migration assay was conducted using transwell chamber. (a) The number of migrated cells that penetrated transwell chamber was expressed as the mean number of cells in the 5 random fields identified within (*P < 0.01 vs. controls). The results shown here were for one representative experiment of three with similar results; (b) Cells on the lower surface of the filter were photographed.
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Figure 4: The effect of GPR34 on migration of HGC-27 cells In vitro. Migration assay was conducted using transwell chamber. (a) The number of migrated cells that penetrated transwell chamber was expressed as the mean number of cells in the 5 random fields identified within (*P < 0.01 vs. controls). The results shown here were for one representative experiment of three with similar results; (b) Cells on the lower surface of the filter were photographed.

Mentions: To evaluate the function of GPR34 on HGC-27 cells migration, transwell chambers were utilized. We found that GPR34 knockdown led to a significantly decreased migratory ability compared with the control group (P < 0.01, Figure 4).


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)

The effect of GPR34 on migration of HGC-27 cells In vitro. Migration assay was conducted using transwell chamber. (a) The number of migrated cells that penetrated transwell chamber was expressed as the mean number of cells in the 5 random fields identified within (*P < 0.01 vs. controls). The results shown here were for one representative experiment of three with similar results; (b) Cells on the lower surface of the filter were photographed.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: The effect of GPR34 on migration of HGC-27 cells In vitro. Migration assay was conducted using transwell chamber. (a) The number of migrated cells that penetrated transwell chamber was expressed as the mean number of cells in the 5 random fields identified within (*P < 0.01 vs. controls). The results shown here were for one representative experiment of three with similar results; (b) Cells on the lower surface of the filter were photographed.
Mentions: To evaluate the function of GPR34 on HGC-27 cells migration, transwell chambers were utilized. We found that GPR34 knockdown led to a significantly decreased migratory ability compared with the control group (P < 0.01, Figure 4).

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