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Suppression of Human Liver Cancer Cell Migration and Invasion via the GABAA Receptor.

Chen ZA, Bao MY, Xu YF, Zha RP, Shi HB, Chen TY, He XH - Cancer Biol Med (2012)

Bottom Line: The effects of GABA on liver cancer cell cytoskeletal were determined by immunofluorescence staining.GABA inhibited human liver cancer cell migration and invasion via the ionotropic GABAA receptor as a result of the induction of liver cancer cell cytoskeletal reorganization.Pretreatment with GABA also significantly reduced intrahepatic liver metastasis and primary tumor formation in vivo.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200032, China.

ABSTRACT

Objective: To investigate the roles of the γ-aminobutyric acid (GABA) in the metastasis of hepatocellular carcinoma (HCC) and to explore the potential of a novel therapeutic approach for the treatment of HCC.

Methods: The expression levels of GABA receptor subunit genes in various HCC cell lines and patients' tissues were detected by quantitative real-time polymerase chain reaction and Western blot analysis. Transwell cell migration and invasion assays were carried out for functional analysis. The effects of GABA on liver cancer cell cytoskeletal were determined by immunofluorescence staining. And the effects of GABA on HCC metastasis in nude mice were evaluated using an in vivo orthotopic model of liver cancer.

Results: The mRNA level of GABA receptor subunits varied between the primary hepatocellular carcinoma tissue and the adjacent non-tumor liver tissue. GABA inhibited human liver cancer cell migration and invasion via the ionotropic GABAA receptor as a result of the induction of liver cancer cell cytoskeletal reorganization. Pretreatment with GABA also significantly reduced intrahepatic liver metastasis and primary tumor formation in vivo.

Conclusions: These findings introduce a potential and novel therapeutic approach for the treatment of cancer patients based on the modulation of the GABAergic system.

No MeSH data available.


Related in: MedlinePlus

GABA-inhibited HCC migration and invasion via GABAA receptor. A, B: Migration and invasion assays were performed in SK-Hep1 and SMMC-7721 cells stimulated with 10 µM GABAA or GABAB receptor agonists for 24 h. Bac, baclofen. The results show the quantification of 5 randomly selected fields. C, D: Migration and invasion assays were performed in SK-Hep1 and SMMC-7721 cells stimulated with 10 µM GABA for 24 h with or without 2 h pre-incubation with GABAA or GABAB receptors antagonists. CGP, CGP 35348 hydrate. Data are representative of three independent experiments and are shown as means ±SEM (*, P<0.05, **, P<0.01, by Student’s t-test).
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f3: GABA-inhibited HCC migration and invasion via GABAA receptor. A, B: Migration and invasion assays were performed in SK-Hep1 and SMMC-7721 cells stimulated with 10 µM GABAA or GABAB receptor agonists for 24 h. Bac, baclofen. The results show the quantification of 5 randomly selected fields. C, D: Migration and invasion assays were performed in SK-Hep1 and SMMC-7721 cells stimulated with 10 µM GABA for 24 h with or without 2 h pre-incubation with GABAA or GABAB receptors antagonists. CGP, CGP 35348 hydrate. Data are representative of three independent experiments and are shown as means ±SEM (*, P<0.05, **, P<0.01, by Student’s t-test).

Mentions: The effects of different agonists and antagonists were compared to identify a potential target receptor that is involved in the GABA-mediated inhibition of HCC cell migration and invasion. After administration of specific agonists for the GABAA and GABAB receptors in serum-free culture conditions, a marked decrease was observed in the number of migratory and invasive cells when the cells were incubated with GABAA receptor agonists (T101), but not with GABAB receptor agonists (Baclofen) (Figure 3A and 3B). A decrease in the number of migratory and invasive cells was also found with muscimol, a selective GABAA receptor agonist (data not shown). This finding was further supported when the cells were treated with S106 and CGP 35348 hydrate. When S106, a general GABAA receptor antagonist, was added during incubation with GABA, GABA-mediated inhibition was suppressed. However, with CGP 35348 hydrate, a GABAB receptor antagonist, the number of migratory and invasive cells decreased after incubation with GABA (Figure 3C and 3D). Similar results were observed with B7561, a selective GABAA receptor antagonist, and with 2-hydroxysaclofenwere, a GABAB receptor antagonist (data not shown). In sum, the inhibition of GABA on the migration and the invasion of liver cancer cells is mediated by GABAA receptors.


Suppression of Human Liver Cancer Cell Migration and Invasion via the GABAA Receptor.

Chen ZA, Bao MY, Xu YF, Zha RP, Shi HB, Chen TY, He XH - Cancer Biol Med (2012)

GABA-inhibited HCC migration and invasion via GABAA receptor. A, B: Migration and invasion assays were performed in SK-Hep1 and SMMC-7721 cells stimulated with 10 µM GABAA or GABAB receptor agonists for 24 h. Bac, baclofen. The results show the quantification of 5 randomly selected fields. C, D: Migration and invasion assays were performed in SK-Hep1 and SMMC-7721 cells stimulated with 10 µM GABA for 24 h with or without 2 h pre-incubation with GABAA or GABAB receptors antagonists. CGP, CGP 35348 hydrate. Data are representative of three independent experiments and are shown as means ±SEM (*, P<0.05, **, P<0.01, by Student’s t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3643652&req=5

f3: GABA-inhibited HCC migration and invasion via GABAA receptor. A, B: Migration and invasion assays were performed in SK-Hep1 and SMMC-7721 cells stimulated with 10 µM GABAA or GABAB receptor agonists for 24 h. Bac, baclofen. The results show the quantification of 5 randomly selected fields. C, D: Migration and invasion assays were performed in SK-Hep1 and SMMC-7721 cells stimulated with 10 µM GABA for 24 h with or without 2 h pre-incubation with GABAA or GABAB receptors antagonists. CGP, CGP 35348 hydrate. Data are representative of three independent experiments and are shown as means ±SEM (*, P<0.05, **, P<0.01, by Student’s t-test).
Mentions: The effects of different agonists and antagonists were compared to identify a potential target receptor that is involved in the GABA-mediated inhibition of HCC cell migration and invasion. After administration of specific agonists for the GABAA and GABAB receptors in serum-free culture conditions, a marked decrease was observed in the number of migratory and invasive cells when the cells were incubated with GABAA receptor agonists (T101), but not with GABAB receptor agonists (Baclofen) (Figure 3A and 3B). A decrease in the number of migratory and invasive cells was also found with muscimol, a selective GABAA receptor agonist (data not shown). This finding was further supported when the cells were treated with S106 and CGP 35348 hydrate. When S106, a general GABAA receptor antagonist, was added during incubation with GABA, GABA-mediated inhibition was suppressed. However, with CGP 35348 hydrate, a GABAB receptor antagonist, the number of migratory and invasive cells decreased after incubation with GABA (Figure 3C and 3D). Similar results were observed with B7561, a selective GABAA receptor antagonist, and with 2-hydroxysaclofenwere, a GABAB receptor antagonist (data not shown). In sum, the inhibition of GABA on the migration and the invasion of liver cancer cells is mediated by GABAA receptors.

Bottom Line: The effects of GABA on liver cancer cell cytoskeletal were determined by immunofluorescence staining.GABA inhibited human liver cancer cell migration and invasion via the ionotropic GABAA receptor as a result of the induction of liver cancer cell cytoskeletal reorganization.Pretreatment with GABA also significantly reduced intrahepatic liver metastasis and primary tumor formation in vivo.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200032, China.

ABSTRACT

Objective: To investigate the roles of the γ-aminobutyric acid (GABA) in the metastasis of hepatocellular carcinoma (HCC) and to explore the potential of a novel therapeutic approach for the treatment of HCC.

Methods: The expression levels of GABA receptor subunit genes in various HCC cell lines and patients' tissues were detected by quantitative real-time polymerase chain reaction and Western blot analysis. Transwell cell migration and invasion assays were carried out for functional analysis. The effects of GABA on liver cancer cell cytoskeletal were determined by immunofluorescence staining. And the effects of GABA on HCC metastasis in nude mice were evaluated using an in vivo orthotopic model of liver cancer.

Results: The mRNA level of GABA receptor subunits varied between the primary hepatocellular carcinoma tissue and the adjacent non-tumor liver tissue. GABA inhibited human liver cancer cell migration and invasion via the ionotropic GABAA receptor as a result of the induction of liver cancer cell cytoskeletal reorganization. Pretreatment with GABA also significantly reduced intrahepatic liver metastasis and primary tumor formation in vivo.

Conclusions: These findings introduce a potential and novel therapeutic approach for the treatment of cancer patients based on the modulation of the GABAergic system.

No MeSH data available.


Related in: MedlinePlus