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MicroRNA-24 inhibits growth, induces apoptosis, and reverses radioresistance in laryngeal squamous cell carcinoma by targeting X-linked inhibitor of apoptosis protein.

Xu L, Chen Z, Xue F, Chen W, Ma R, Cheng S, Cui P - Cancer Cell Int. (2015)

Bottom Line: Functional analyses indicated that re-expression of miR-24 inhibits growth, reduces colony formation, and enhances apoptosis in LSCC cells.Upregulation of miR-24 inhibits XIAP protein expression in LSCC cells, and silencing of XIAP mimics the effects of miR-24 upregulation on LSCC cells.In addition, XIAP mRNA expression significantly increases in LSCC tissues and is inversely correlated with miR-24 expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology-Head and Neck Surgery, Tangdu Hospital and Laboratory for Laryngotracheal Reconstruction, Fourth Military Medical University, Xi'an, Shaanxi 710038 PR China ; Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Command, Nanjing, Jiangsu 210002 PR China.

ABSTRACT

Background: Increasing evidence indicates that dysregulation of microRNAs is involved in tumor progression and development. The aim of this study was to investigate the expression of microRNA-24 (miR-24) and its function in laryngeal squamous cell carcinoma (LSCC).

Methods: Quantitative RT-PCR (qRT-PCR) was used to detect miR-24 expression in LSCC cell lines and tissue samples. MTT, colony formation, and flow cytometry was performed to analyze the effects of miR-24 expression on growth, apoptosis, and radiosensitivity of LSCC cells. Dual-luciferase reporter assays were performed to examine regulation of putative miR-24 targets. Expression of X-linked inhibitor of apoptosis protein (XIAP) mRNA and protein, cleaved or total caspase-3, and cleaved or total PARP protein were detected by qRT-PCR and western blotting assays, respectively.

Results: miR-24 expression levels in LSCC cell lines or tissue were significantly lower than in a normal human keratinocyte cell line or adjacent normal tissues. Functional analyses indicated that re-expression of miR-24 inhibits growth, reduces colony formation, and enhances apoptosis in LSCC cells. In addition, miR-24 upregulation increases LSCC sensitivity to irradiation by enhancing irradiation-induced apoptosis, and luciferase activity indicated that miR-24 binds to the 3'-untranslated region (3'-UTR) of XIAP mRNA. Upregulation of miR-24 inhibits XIAP protein expression in LSCC cells, and silencing of XIAP mimics the effects of miR-24 upregulation on LSCC cells. In addition, XIAP mRNA expression significantly increases in LSCC tissues and is inversely correlated with miR-24 expression.

Conclusions: Our data suggest that miR-24 inhibits growth, increases apoptosis, and enhances radiosensitivity in LSCC cells by targeting XIAP. Therefore, miR-24 may be a potential molecular target for the treatment of human LSCC.

No MeSH data available.


Related in: MedlinePlus

XIAP expression is upregulated in LSCC cells and tissues and inversely correlated with miR-24 expression. a qRT-PCR detection of XIAP mRNA expression in Hep-2 and AMC-HN-8, and HaCaT cells. GAPDH was used as an internal control. b qRT-PCR detection of XIAP mRNA expression in 15 paired LSCC tissues and adjacent normal tissues. GAPDH was used as an internal control. T: LSCC tissues; N: the adjacent normal tissues. c Statistical analysis reveals an inverse correlation between relative miR-24 and XIAP mRNA expression level in LSCC tissues (n = 15; r = −0.508; P < 0.001). Corresponding P values analyzed by Spearman correlation test are indicated. **P < 0.01 vs control
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Fig7: XIAP expression is upregulated in LSCC cells and tissues and inversely correlated with miR-24 expression. a qRT-PCR detection of XIAP mRNA expression in Hep-2 and AMC-HN-8, and HaCaT cells. GAPDH was used as an internal control. b qRT-PCR detection of XIAP mRNA expression in 15 paired LSCC tissues and adjacent normal tissues. GAPDH was used as an internal control. T: LSCC tissues; N: the adjacent normal tissues. c Statistical analysis reveals an inverse correlation between relative miR-24 and XIAP mRNA expression level in LSCC tissues (n = 15; r = −0.508; P < 0.001). Corresponding P values analyzed by Spearman correlation test are indicated. **P < 0.01 vs control

Mentions: We further examined expression of XIAP mRNA in HaCaT and Hep-2 and AMC-HN-8 cells by qRT-PCR, and found that the expression of XIAP mRNA in HaCaT cell line was lower than that in the LSCC cell lines (Fig. 7a). In addition, expression of XIAP mRNA in 15 paired LSCC and adjacent normal tissues showed that XIAP mRNA in LSCC tissues was elevated (P < 0.001; Fig. 7b). We then observed that expression level of XIAP mRNA expression levels inversely correlated with the expression level of miR-24 in LSCC tissues (Pearson’s correlation, r = −0.508; P < 0.001; Fig. 7c). Therefore, the increased XIAP mRNA expression in LSCC tissues correlates with low-level miR-24 expression.Fig. 7


MicroRNA-24 inhibits growth, induces apoptosis, and reverses radioresistance in laryngeal squamous cell carcinoma by targeting X-linked inhibitor of apoptosis protein.

Xu L, Chen Z, Xue F, Chen W, Ma R, Cheng S, Cui P - Cancer Cell Int. (2015)

XIAP expression is upregulated in LSCC cells and tissues and inversely correlated with miR-24 expression. a qRT-PCR detection of XIAP mRNA expression in Hep-2 and AMC-HN-8, and HaCaT cells. GAPDH was used as an internal control. b qRT-PCR detection of XIAP mRNA expression in 15 paired LSCC tissues and adjacent normal tissues. GAPDH was used as an internal control. T: LSCC tissues; N: the adjacent normal tissues. c Statistical analysis reveals an inverse correlation between relative miR-24 and XIAP mRNA expression level in LSCC tissues (n = 15; r = −0.508; P < 0.001). Corresponding P values analyzed by Spearman correlation test are indicated. **P < 0.01 vs control
© Copyright Policy - open-access
Related In: Results  -  Collection

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Fig7: XIAP expression is upregulated in LSCC cells and tissues and inversely correlated with miR-24 expression. a qRT-PCR detection of XIAP mRNA expression in Hep-2 and AMC-HN-8, and HaCaT cells. GAPDH was used as an internal control. b qRT-PCR detection of XIAP mRNA expression in 15 paired LSCC tissues and adjacent normal tissues. GAPDH was used as an internal control. T: LSCC tissues; N: the adjacent normal tissues. c Statistical analysis reveals an inverse correlation between relative miR-24 and XIAP mRNA expression level in LSCC tissues (n = 15; r = −0.508; P < 0.001). Corresponding P values analyzed by Spearman correlation test are indicated. **P < 0.01 vs control
Mentions: We further examined expression of XIAP mRNA in HaCaT and Hep-2 and AMC-HN-8 cells by qRT-PCR, and found that the expression of XIAP mRNA in HaCaT cell line was lower than that in the LSCC cell lines (Fig. 7a). In addition, expression of XIAP mRNA in 15 paired LSCC and adjacent normal tissues showed that XIAP mRNA in LSCC tissues was elevated (P < 0.001; Fig. 7b). We then observed that expression level of XIAP mRNA expression levels inversely correlated with the expression level of miR-24 in LSCC tissues (Pearson’s correlation, r = −0.508; P < 0.001; Fig. 7c). Therefore, the increased XIAP mRNA expression in LSCC tissues correlates with low-level miR-24 expression.Fig. 7

Bottom Line: Functional analyses indicated that re-expression of miR-24 inhibits growth, reduces colony formation, and enhances apoptosis in LSCC cells.Upregulation of miR-24 inhibits XIAP protein expression in LSCC cells, and silencing of XIAP mimics the effects of miR-24 upregulation on LSCC cells.In addition, XIAP mRNA expression significantly increases in LSCC tissues and is inversely correlated with miR-24 expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Otolaryngology-Head and Neck Surgery, Tangdu Hospital and Laboratory for Laryngotracheal Reconstruction, Fourth Military Medical University, Xi'an, Shaanxi 710038 PR China ; Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Command, Nanjing, Jiangsu 210002 PR China.

ABSTRACT

Background: Increasing evidence indicates that dysregulation of microRNAs is involved in tumor progression and development. The aim of this study was to investigate the expression of microRNA-24 (miR-24) and its function in laryngeal squamous cell carcinoma (LSCC).

Methods: Quantitative RT-PCR (qRT-PCR) was used to detect miR-24 expression in LSCC cell lines and tissue samples. MTT, colony formation, and flow cytometry was performed to analyze the effects of miR-24 expression on growth, apoptosis, and radiosensitivity of LSCC cells. Dual-luciferase reporter assays were performed to examine regulation of putative miR-24 targets. Expression of X-linked inhibitor of apoptosis protein (XIAP) mRNA and protein, cleaved or total caspase-3, and cleaved or total PARP protein were detected by qRT-PCR and western blotting assays, respectively.

Results: miR-24 expression levels in LSCC cell lines or tissue were significantly lower than in a normal human keratinocyte cell line or adjacent normal tissues. Functional analyses indicated that re-expression of miR-24 inhibits growth, reduces colony formation, and enhances apoptosis in LSCC cells. In addition, miR-24 upregulation increases LSCC sensitivity to irradiation by enhancing irradiation-induced apoptosis, and luciferase activity indicated that miR-24 binds to the 3'-untranslated region (3'-UTR) of XIAP mRNA. Upregulation of miR-24 inhibits XIAP protein expression in LSCC cells, and silencing of XIAP mimics the effects of miR-24 upregulation on LSCC cells. In addition, XIAP mRNA expression significantly increases in LSCC tissues and is inversely correlated with miR-24 expression.

Conclusions: Our data suggest that miR-24 inhibits growth, increases apoptosis, and enhances radiosensitivity in LSCC cells by targeting XIAP. Therefore, miR-24 may be a potential molecular target for the treatment of human LSCC.

No MeSH data available.


Related in: MedlinePlus