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HOXB1 Is a Tumor Suppressor Gene Regulated by miR-3175 in Glioma.

Han L, Liu D, Li Z, Tian N, Han Z, Wang G, Fu Y, Guo Z, Zhu Z, Du C, Tian Y - PLoS ONE (2015)

Bottom Line: The HOXB1 gene plays a critical role as an oncogene in diverse tumors.We show that HOXB1 expression is significantly downregulated in glioma tissues and cell lines, and that its expression may be closely associated with the degree of malignancy.Our results suggest that HOXB1 functions as a tumor suppressor, regulated by miR-3175 in glioma.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.

ABSTRACT
The HOXB1 gene plays a critical role as an oncogene in diverse tumors. However, the functional role of HOXB1 and the mechanism regulating HOXB1 expression in glioma are not fully understood. A preliminary bioinformatics analysis showed that HOXB1 is ectopically expressed in glioma, and that HOXB1 is a possible target of miR-3175. In this study, we investigated the function of HOXB1 and the relationship between HOXB1 and miR-3175 in glioma. We show that HOXB1 expression is significantly downregulated in glioma tissues and cell lines, and that its expression may be closely associated with the degree of malignancy. Reduced HOXB1 expression promoted the proliferation and invasion of glioma cells, and inhibited their apoptosis in vitro, and the downregulation of HOXB1 was also associated with worse survival in glioma patients. More importantly, HOXB1 was shown experimentally to be a direct target of miR-3175 in this study. The downregulated expression of miR-3175 inhibited cell proliferation and invasion, and promoted apoptosis in glioma. The oncogenicity induced by low HOXB1 expression was prevented by an miR-3175 inhibitor in glioma cells. Our results suggest that HOXB1 functions as a tumor suppressor, regulated by miR-3175 in glioma. These results clarify the pathogenesis of glioma and offer a potential target for its treatment.

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Oncogenicity induced by low HOXB1 expression is prevented by miR-3175 inhibitor in glioma cells.(A) An MTT cell proliferation assay was performed at 48 h after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC in glioma cell lines. (B-C) Flow cytometric analysis of apoptosis in glioma cell lines after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC. Apoptosis was assessed with Annexin V-FITC/PI. (D-E) Western blot analysis of several apoptosis-related proteins (procaspase-3, p53 and cytochrome c) expression in U87 cells after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC. All results are representative of three independent experiments, and a statistical analysis is performed (mean ± SD, *P < 0.05, **P < 0.01, and ***P < 0.001).
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pone.0142387.g006: Oncogenicity induced by low HOXB1 expression is prevented by miR-3175 inhibitor in glioma cells.(A) An MTT cell proliferation assay was performed at 48 h after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC in glioma cell lines. (B-C) Flow cytometric analysis of apoptosis in glioma cell lines after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC. Apoptosis was assessed with Annexin V-FITC/PI. (D-E) Western blot analysis of several apoptosis-related proteins (procaspase-3, p53 and cytochrome c) expression in U87 cells after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC. All results are representative of three independent experiments, and a statistical analysis is performed (mean ± SD, *P < 0.05, **P < 0.01, and ***P < 0.001).

Mentions: As shown above, we found that HOXB1 is a direct target of miR-3175 and HOXB1 expression was significantly reduced after transfection with the miR-3175 mimic. We also demonstrated that the reduced expression of HOXB1 promoted cell proliferation and invasion and inhibited apoptosis, similar to the phenomena induced by miR-3175. The miR-3175 inhibitor was shown to inhibit the proliferation and invasion of glioma cells and to promote cell apoptosis in glioma. Cell proliferation and apoptosis assays were performed to clarify whether the oncogenicity induced by low HOXB1 expression was affected by miR-3175 in glioma, after cells were cotransfected with si-HOXB1, the miR-3175 inhibitor, or the corresponding NCs. The results showed that the miR-3175 inhibitor significantly prevented the increased cell proliferation and suppressed the apoptosis induced by low HOXB1 expression (Fig 6A–6C), indicating strongly that miR-3175 is a critical participant in the tumor-suppressor activity of HOXB1 in glioma. A western blot analysis of several downstream apoptosis-related proteins (procaspase-3, p53, and cytochrome c) was performed to preliminarily investigate the signaling pathway affecting HOXB1-induced apoptosis, which is dysregulated in the pathogenesis of glioma. This analysis showed that procaspase-3, p53, and cytochrome c are involved in HOXB1-induced apoptosis (Fig 6D and 6E).


HOXB1 Is a Tumor Suppressor Gene Regulated by miR-3175 in Glioma.

Han L, Liu D, Li Z, Tian N, Han Z, Wang G, Fu Y, Guo Z, Zhu Z, Du C, Tian Y - PLoS ONE (2015)

Oncogenicity induced by low HOXB1 expression is prevented by miR-3175 inhibitor in glioma cells.(A) An MTT cell proliferation assay was performed at 48 h after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC in glioma cell lines. (B-C) Flow cytometric analysis of apoptosis in glioma cell lines after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC. Apoptosis was assessed with Annexin V-FITC/PI. (D-E) Western blot analysis of several apoptosis-related proteins (procaspase-3, p53 and cytochrome c) expression in U87 cells after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC. All results are representative of three independent experiments, and a statistical analysis is performed (mean ± SD, *P < 0.05, **P < 0.01, and ***P < 0.001).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4643923&req=5

pone.0142387.g006: Oncogenicity induced by low HOXB1 expression is prevented by miR-3175 inhibitor in glioma cells.(A) An MTT cell proliferation assay was performed at 48 h after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC in glioma cell lines. (B-C) Flow cytometric analysis of apoptosis in glioma cell lines after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC. Apoptosis was assessed with Annexin V-FITC/PI. (D-E) Western blot analysis of several apoptosis-related proteins (procaspase-3, p53 and cytochrome c) expression in U87 cells after transfection with si-NC + inhibitor NC, si-HOXB1 + miR-3175 inhibitor and si-HOXB1 + inhibitor NC. All results are representative of three independent experiments, and a statistical analysis is performed (mean ± SD, *P < 0.05, **P < 0.01, and ***P < 0.001).
Mentions: As shown above, we found that HOXB1 is a direct target of miR-3175 and HOXB1 expression was significantly reduced after transfection with the miR-3175 mimic. We also demonstrated that the reduced expression of HOXB1 promoted cell proliferation and invasion and inhibited apoptosis, similar to the phenomena induced by miR-3175. The miR-3175 inhibitor was shown to inhibit the proliferation and invasion of glioma cells and to promote cell apoptosis in glioma. Cell proliferation and apoptosis assays were performed to clarify whether the oncogenicity induced by low HOXB1 expression was affected by miR-3175 in glioma, after cells were cotransfected with si-HOXB1, the miR-3175 inhibitor, or the corresponding NCs. The results showed that the miR-3175 inhibitor significantly prevented the increased cell proliferation and suppressed the apoptosis induced by low HOXB1 expression (Fig 6A–6C), indicating strongly that miR-3175 is a critical participant in the tumor-suppressor activity of HOXB1 in glioma. A western blot analysis of several downstream apoptosis-related proteins (procaspase-3, p53, and cytochrome c) was performed to preliminarily investigate the signaling pathway affecting HOXB1-induced apoptosis, which is dysregulated in the pathogenesis of glioma. This analysis showed that procaspase-3, p53, and cytochrome c are involved in HOXB1-induced apoptosis (Fig 6D and 6E).

Bottom Line: The HOXB1 gene plays a critical role as an oncogene in diverse tumors.We show that HOXB1 expression is significantly downregulated in glioma tissues and cell lines, and that its expression may be closely associated with the degree of malignancy.Our results suggest that HOXB1 functions as a tumor suppressor, regulated by miR-3175 in glioma.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosurgery, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.

ABSTRACT
The HOXB1 gene plays a critical role as an oncogene in diverse tumors. However, the functional role of HOXB1 and the mechanism regulating HOXB1 expression in glioma are not fully understood. A preliminary bioinformatics analysis showed that HOXB1 is ectopically expressed in glioma, and that HOXB1 is a possible target of miR-3175. In this study, we investigated the function of HOXB1 and the relationship between HOXB1 and miR-3175 in glioma. We show that HOXB1 expression is significantly downregulated in glioma tissues and cell lines, and that its expression may be closely associated with the degree of malignancy. Reduced HOXB1 expression promoted the proliferation and invasion of glioma cells, and inhibited their apoptosis in vitro, and the downregulation of HOXB1 was also associated with worse survival in glioma patients. More importantly, HOXB1 was shown experimentally to be a direct target of miR-3175 in this study. The downregulated expression of miR-3175 inhibited cell proliferation and invasion, and promoted apoptosis in glioma. The oncogenicity induced by low HOXB1 expression was prevented by an miR-3175 inhibitor in glioma cells. Our results suggest that HOXB1 functions as a tumor suppressor, regulated by miR-3175 in glioma. These results clarify the pathogenesis of glioma and offer a potential target for its treatment.

Show MeSH
Related in: MedlinePlus