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Decreased expression of Beclin‑1 is significantly associated with a poor prognosis in oral tongue squamous cell carcinoma.

Hu Z, Zhong Z, Huang S, Wen H, Chen X, Chu H, Li Q, Sun C - Mol Med Rep (2016)

Bottom Line: Furthermore, MTT and colony formation assays were performed to investigate the effect of Beclin‑1 on the proliferation and clonogenicity of OTSCC cells.It was demonstrated that Beclin‑1 expression was significantly decreased in the majority of the 14 OTSCC tissues and the 5 OTSCC cell lines relative to the matched non‑cancerous tissues and the normal tongue epithelial cell line, respectively.The in vitro assays indicated that the overexpression of Beclin‑1 significantly inhibits the proliferation and clonogenicity of OTSCC cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Head and Neck Surgery, The Third Affiliated Hospital of Kunming Medical University, Kunming, Yunnan 650118, P.R. China.

ABSTRACT
The autophagy-related gene Beclin-1 is critical in the regulation of tumourigenesis and progression, but its role in oral tongue squamous cell carcinoma (OTSCC) has not yet been reported. This study aimed to investigate Beclin‑1 expression and its significance in OTSCC. Beclin‑1 expression was assessed by reverse transcription‑quantitative polymerase chain reaction or western blot analysis in 14 OTSCC tissues and matched adjacent noncancerous tissues as well as in 5 OTSCC cell lines and a normal tongue epithelial cell line. Beclin‑1 protein expression was examined by immunohistochemistry in 133 OTSCC specimens, and the correlation between Beclin‑1 expression and clinicopathological features was investigated. Furthermore, MTT and colony formation assays were performed to investigate the effect of Beclin‑1 on the proliferation and clonogenicity of OTSCC cells. It was demonstrated that Beclin‑1 expression was significantly decreased in the majority of the 14 OTSCC tissues and the 5 OTSCC cell lines relative to the matched non‑cancerous tissues and the normal tongue epithelial cell line, respectively. Immunohistochemistry analysis revealed that decreased Beclin‑1 expression was significantly correlated with poor differentiation, lymph node metastasis, advanced clinical tumour‑node‑metastasis stage, and a poor prognosis in patients with OTSCC. The in vitro assays indicated that the overexpression of Beclin‑1 significantly inhibits the proliferation and clonogenicity of OTSCC cells. These results demonstrate that Beclin‑1 acts as a tumour suppressor in the development or progression of OTSCC and that Beclin‑1 may represent a novel prognostic marker for patients with OTSCC.

No MeSH data available.


Related in: MedlinePlus

Beclin-1 overexpression inhibits the proliferation and transforma-overexpression inhibits the proliferation and transformation of OTSCC cells. (A) Beclin-1 inhibits the cell proliferation of OTSCC as demonstrated by an MTT assay. (B) Beclin-1 inhibits the transformation of OTSCC cells as demonstrated by a colony formation assay. (C) Beclin-1 inhibits the transformation of OTSCC cells as demonstrated by the soft-agar anchorage-independent growth assay. OTSCC, oral tongue squamous cell carcinoma. Magnification, ×200. ***P<0.001 compared with the control.
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f5-mmr-14-02-1567: Beclin-1 overexpression inhibits the proliferation and transforma-overexpression inhibits the proliferation and transformation of OTSCC cells. (A) Beclin-1 inhibits the cell proliferation of OTSCC as demonstrated by an MTT assay. (B) Beclin-1 inhibits the transformation of OTSCC cells as demonstrated by a colony formation assay. (C) Beclin-1 inhibits the transformation of OTSCC cells as demonstrated by the soft-agar anchorage-independent growth assay. OTSCC, oral tongue squamous cell carcinoma. Magnification, ×200. ***P<0.001 compared with the control.

Mentions: Beclin-1 is a pivotal gene in the process of autophagy. Therefore, the influence of Beclin-1 overexpression on autophagy upon starvation of OTSCC cells was investigated. As normal culture medium contains serum which may influence autophagy level, the cells were cultured in Earle's balanced salts (EBSS) to observe autophagy level. No significant difference in the expression levels of LC3-II and p62 was identified between the Beclin-1 overexpression cell line and the vehicle cell line subjected to starvation by culturing in EBSS for 1, 2, 4 or 8 h. The number of GFP-LC3 puncta was also counted in at least 50 cells after transient transfection of pEGF-C2-LC3, and no significant difference was observed between the cells overexpressing Beclin-1 and the vehicle group (12.66±0.32 vs. 11.98±0.34; P=0.7996) (Fig. 4). To determine whether Beclin-1 inhibited OTSCC cell proliferation, an MTT assay was performed, and the results suggested that Beclin-1 overexpression significantly inhibited the proliferation of OTSCC cell lines (Fig. 5A).


Decreased expression of Beclin‑1 is significantly associated with a poor prognosis in oral tongue squamous cell carcinoma.

Hu Z, Zhong Z, Huang S, Wen H, Chen X, Chu H, Li Q, Sun C - Mol Med Rep (2016)

Beclin-1 overexpression inhibits the proliferation and transforma-overexpression inhibits the proliferation and transformation of OTSCC cells. (A) Beclin-1 inhibits the cell proliferation of OTSCC as demonstrated by an MTT assay. (B) Beclin-1 inhibits the transformation of OTSCC cells as demonstrated by a colony formation assay. (C) Beclin-1 inhibits the transformation of OTSCC cells as demonstrated by the soft-agar anchorage-independent growth assay. OTSCC, oral tongue squamous cell carcinoma. Magnification, ×200. ***P<0.001 compared with the control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940089&req=5

f5-mmr-14-02-1567: Beclin-1 overexpression inhibits the proliferation and transforma-overexpression inhibits the proliferation and transformation of OTSCC cells. (A) Beclin-1 inhibits the cell proliferation of OTSCC as demonstrated by an MTT assay. (B) Beclin-1 inhibits the transformation of OTSCC cells as demonstrated by a colony formation assay. (C) Beclin-1 inhibits the transformation of OTSCC cells as demonstrated by the soft-agar anchorage-independent growth assay. OTSCC, oral tongue squamous cell carcinoma. Magnification, ×200. ***P<0.001 compared with the control.
Mentions: Beclin-1 is a pivotal gene in the process of autophagy. Therefore, the influence of Beclin-1 overexpression on autophagy upon starvation of OTSCC cells was investigated. As normal culture medium contains serum which may influence autophagy level, the cells were cultured in Earle's balanced salts (EBSS) to observe autophagy level. No significant difference in the expression levels of LC3-II and p62 was identified between the Beclin-1 overexpression cell line and the vehicle cell line subjected to starvation by culturing in EBSS for 1, 2, 4 or 8 h. The number of GFP-LC3 puncta was also counted in at least 50 cells after transient transfection of pEGF-C2-LC3, and no significant difference was observed between the cells overexpressing Beclin-1 and the vehicle group (12.66±0.32 vs. 11.98±0.34; P=0.7996) (Fig. 4). To determine whether Beclin-1 inhibited OTSCC cell proliferation, an MTT assay was performed, and the results suggested that Beclin-1 overexpression significantly inhibited the proliferation of OTSCC cell lines (Fig. 5A).

Bottom Line: Furthermore, MTT and colony formation assays were performed to investigate the effect of Beclin‑1 on the proliferation and clonogenicity of OTSCC cells.It was demonstrated that Beclin‑1 expression was significantly decreased in the majority of the 14 OTSCC tissues and the 5 OTSCC cell lines relative to the matched non‑cancerous tissues and the normal tongue epithelial cell line, respectively.The in vitro assays indicated that the overexpression of Beclin‑1 significantly inhibits the proliferation and clonogenicity of OTSCC cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Head and Neck Surgery, The Third Affiliated Hospital of Kunming Medical University, Kunming, Yunnan 650118, P.R. China.

ABSTRACT
The autophagy-related gene Beclin-1 is critical in the regulation of tumourigenesis and progression, but its role in oral tongue squamous cell carcinoma (OTSCC) has not yet been reported. This study aimed to investigate Beclin‑1 expression and its significance in OTSCC. Beclin‑1 expression was assessed by reverse transcription‑quantitative polymerase chain reaction or western blot analysis in 14 OTSCC tissues and matched adjacent noncancerous tissues as well as in 5 OTSCC cell lines and a normal tongue epithelial cell line. Beclin‑1 protein expression was examined by immunohistochemistry in 133 OTSCC specimens, and the correlation between Beclin‑1 expression and clinicopathological features was investigated. Furthermore, MTT and colony formation assays were performed to investigate the effect of Beclin‑1 on the proliferation and clonogenicity of OTSCC cells. It was demonstrated that Beclin‑1 expression was significantly decreased in the majority of the 14 OTSCC tissues and the 5 OTSCC cell lines relative to the matched non‑cancerous tissues and the normal tongue epithelial cell line, respectively. Immunohistochemistry analysis revealed that decreased Beclin‑1 expression was significantly correlated with poor differentiation, lymph node metastasis, advanced clinical tumour‑node‑metastasis stage, and a poor prognosis in patients with OTSCC. The in vitro assays indicated that the overexpression of Beclin‑1 significantly inhibits the proliferation and clonogenicity of OTSCC cells. These results demonstrate that Beclin‑1 acts as a tumour suppressor in the development or progression of OTSCC and that Beclin‑1 may represent a novel prognostic marker for patients with OTSCC.

No MeSH data available.


Related in: MedlinePlus