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Gold nano-particles (AuNPs) carrying anti-EBV-miR-BART7-3p inhibit growth of EBV-positive nasopharyngeal carcinoma.

Cai L, Li J, Zhang X, Lu Y, Wang J, Lyu X, Chen Y, Liu J, Cai H, Wang Y, Li X - Oncotarget (2015)

Bottom Line: Based on these results, we conducted a therapeutic experiment by using gold nano-particles (AuNPs) carrying anti-EBV-miR-BART7-3p.Silencing of EBV-miR-BART7-3p reduced tumor growth in animal model.We conclude that EBV-miR-BART7-3p favors carcinogenesis, representing a potential target for miRNA-based therapy.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research Institute, Southern Medical University, Guangzhou 510515, China.

ABSTRACT
Epstein-Barr virus (EBV) infection is a major etiological factor for nasopharyngeal carcinoma (NPC). Several EBV-encoded BART miRNAs have been associated with viral latency, immune escape, cell survival, cell proliferation and apoptosis. Here, we report that EBV-miR-BART7-3p, an EBV-encoded BART miRNA highly expressed in NPC, was correlated with cell-cycle progression in vitro and increased tumor formation in vivo. This viral miRNA stimulated the PTEN/PI3K/Akt pathway and induced c-Myc and c-Jun. Knockdown of PTEN mimicked EBV-miR-BART7-3p-induced tumorigenic phenotype. Based on these results, we conducted a therapeutic experiment by using gold nano-particles (AuNPs) carrying anti-EBV-miR-BART7-3p. Silencing of EBV-miR-BART7-3p reduced tumor growth in animal model. We conclude that EBV-miR-BART7-3p favors carcinogenesis, representing a potential target for miRNA-based therapy.

No MeSH data available.


Related in: MedlinePlus

EBV-miR-BART7-3p stimulated PTEN/PI3K/Akt pathway and its downstream signals(A) The expression levels of Akt, p-Akt, c-Myc, c-Jun, CCND1, CCNE1 and p21 were detected using western blotting in 5-8F-BART7-3p and CNE1-BART7-3p cells or (B) after transfected with anti-miR or anti-C. β-actin served as an internal control. (C) The mRNA expression levels of cell-cycle regulators were analyzed by qPCR in 5-8F-BART7-3p and CNE1-BART7-3p cells before and after transfected with anti-miR. All data were normalized to GAPDH expression and plotted as mean values ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001). (D) The diagram of EBV-miR-BART7-3-mediated signaling pathway for the cell growth and proliferation in NPC.
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Figure 2: EBV-miR-BART7-3p stimulated PTEN/PI3K/Akt pathway and its downstream signals(A) The expression levels of Akt, p-Akt, c-Myc, c-Jun, CCND1, CCNE1 and p21 were detected using western blotting in 5-8F-BART7-3p and CNE1-BART7-3p cells or (B) after transfected with anti-miR or anti-C. β-actin served as an internal control. (C) The mRNA expression levels of cell-cycle regulators were analyzed by qPCR in 5-8F-BART7-3p and CNE1-BART7-3p cells before and after transfected with anti-miR. All data were normalized to GAPDH expression and plotted as mean values ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001). (D) The diagram of EBV-miR-BART7-3-mediated signaling pathway for the cell growth and proliferation in NPC.

Mentions: PTEN/PI3K/Akt constitutes an important pathway modulating multiple biological processes. To address the mechanism of EBV-miR-BART7-3p-mediated phenotypic changes, we conducted western blotting assay to examine phosphorylated Akt (Ser473), a centrally important effector of this pathway. We observed that EBV-miR-BART7-3p enhanced p-Akt expression via suppressing PTEN expression, whereas anti-miR-BART7-3p rescued its expression (Figure 2A and 2B). More interestingly, the expression levels of c-Myc and c-Jun, two transcriptional factors that favor cell growth and proliferation of cancers [27–29], were also increased accordingly (Figure 2A and 2B), indicating that EBV-miR-BART7-3p stimulated PI3K/Akt/c-myc and c-Jun through suppressing PTEN in NPC cells (Figure 2A and 2B). Following the observation of EBV-miR-BART7-3p-mediated growth promotion, we next evaluated the expression of cell-cycle associated genes in EBV-miR-BART7-3p overexpressing or suppressing cells. Two cell-cycle regulators (CCND1 and CCNE1) were highly expressed in EBV-miR-BART7-3p overexpressing cells and lowly expressed in EBV-miR-BART7-3p suppressing cells, whereas P21CIP1, a cell-cycle inhibitor, was lowly expressed in EBV-miR-BART7-3p overexpressing cells and highly expressed in EBV-miR-BART7-3p suppressing cells (Figure 2A and 2B), suggesting the effects of EBV-miR-BART7-3p on cell cycle process probably through activating Akt/c-myc and c-Jun. Furthermore, we did an expanded qPCR examination of cell-cycle regulators and inhibitors. The levels of cyclins and cyclin-dependent kinases were generally increased more than 2-fold upon EBV-miR-BART7-3p overexpression, whereas the levels of cell cycle inhibitors were decreased at least 2-fold (Figure 2C). The opposite results appeared in 5-8F-BART7-3p and CNE1-BART7-3p cells after treating with anti-BART7-3p (Figure 2C). Therefore, these data indicated that EBV-miR-BART7-3p stimulated the PTEN/PI3K/Akt signaling pathway and induced c-Myc and c-Jun, thereby influencing cell cycle process and eventually promoting cell growth and proliferation of NPC (Figure 2D).


Gold nano-particles (AuNPs) carrying anti-EBV-miR-BART7-3p inhibit growth of EBV-positive nasopharyngeal carcinoma.

Cai L, Li J, Zhang X, Lu Y, Wang J, Lyu X, Chen Y, Liu J, Cai H, Wang Y, Li X - Oncotarget (2015)

EBV-miR-BART7-3p stimulated PTEN/PI3K/Akt pathway and its downstream signals(A) The expression levels of Akt, p-Akt, c-Myc, c-Jun, CCND1, CCNE1 and p21 were detected using western blotting in 5-8F-BART7-3p and CNE1-BART7-3p cells or (B) after transfected with anti-miR or anti-C. β-actin served as an internal control. (C) The mRNA expression levels of cell-cycle regulators were analyzed by qPCR in 5-8F-BART7-3p and CNE1-BART7-3p cells before and after transfected with anti-miR. All data were normalized to GAPDH expression and plotted as mean values ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001). (D) The diagram of EBV-miR-BART7-3-mediated signaling pathway for the cell growth and proliferation in NPC.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4480720&req=5

Figure 2: EBV-miR-BART7-3p stimulated PTEN/PI3K/Akt pathway and its downstream signals(A) The expression levels of Akt, p-Akt, c-Myc, c-Jun, CCND1, CCNE1 and p21 were detected using western blotting in 5-8F-BART7-3p and CNE1-BART7-3p cells or (B) after transfected with anti-miR or anti-C. β-actin served as an internal control. (C) The mRNA expression levels of cell-cycle regulators were analyzed by qPCR in 5-8F-BART7-3p and CNE1-BART7-3p cells before and after transfected with anti-miR. All data were normalized to GAPDH expression and plotted as mean values ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001). (D) The diagram of EBV-miR-BART7-3-mediated signaling pathway for the cell growth and proliferation in NPC.
Mentions: PTEN/PI3K/Akt constitutes an important pathway modulating multiple biological processes. To address the mechanism of EBV-miR-BART7-3p-mediated phenotypic changes, we conducted western blotting assay to examine phosphorylated Akt (Ser473), a centrally important effector of this pathway. We observed that EBV-miR-BART7-3p enhanced p-Akt expression via suppressing PTEN expression, whereas anti-miR-BART7-3p rescued its expression (Figure 2A and 2B). More interestingly, the expression levels of c-Myc and c-Jun, two transcriptional factors that favor cell growth and proliferation of cancers [27–29], were also increased accordingly (Figure 2A and 2B), indicating that EBV-miR-BART7-3p stimulated PI3K/Akt/c-myc and c-Jun through suppressing PTEN in NPC cells (Figure 2A and 2B). Following the observation of EBV-miR-BART7-3p-mediated growth promotion, we next evaluated the expression of cell-cycle associated genes in EBV-miR-BART7-3p overexpressing or suppressing cells. Two cell-cycle regulators (CCND1 and CCNE1) were highly expressed in EBV-miR-BART7-3p overexpressing cells and lowly expressed in EBV-miR-BART7-3p suppressing cells, whereas P21CIP1, a cell-cycle inhibitor, was lowly expressed in EBV-miR-BART7-3p overexpressing cells and highly expressed in EBV-miR-BART7-3p suppressing cells (Figure 2A and 2B), suggesting the effects of EBV-miR-BART7-3p on cell cycle process probably through activating Akt/c-myc and c-Jun. Furthermore, we did an expanded qPCR examination of cell-cycle regulators and inhibitors. The levels of cyclins and cyclin-dependent kinases were generally increased more than 2-fold upon EBV-miR-BART7-3p overexpression, whereas the levels of cell cycle inhibitors were decreased at least 2-fold (Figure 2C). The opposite results appeared in 5-8F-BART7-3p and CNE1-BART7-3p cells after treating with anti-BART7-3p (Figure 2C). Therefore, these data indicated that EBV-miR-BART7-3p stimulated the PTEN/PI3K/Akt signaling pathway and induced c-Myc and c-Jun, thereby influencing cell cycle process and eventually promoting cell growth and proliferation of NPC (Figure 2D).

Bottom Line: Based on these results, we conducted a therapeutic experiment by using gold nano-particles (AuNPs) carrying anti-EBV-miR-BART7-3p.Silencing of EBV-miR-BART7-3p reduced tumor growth in animal model.We conclude that EBV-miR-BART7-3p favors carcinogenesis, representing a potential target for miRNA-based therapy.

View Article: PubMed Central - PubMed

Affiliation: Cancer Research Institute, Southern Medical University, Guangzhou 510515, China.

ABSTRACT
Epstein-Barr virus (EBV) infection is a major etiological factor for nasopharyngeal carcinoma (NPC). Several EBV-encoded BART miRNAs have been associated with viral latency, immune escape, cell survival, cell proliferation and apoptosis. Here, we report that EBV-miR-BART7-3p, an EBV-encoded BART miRNA highly expressed in NPC, was correlated with cell-cycle progression in vitro and increased tumor formation in vivo. This viral miRNA stimulated the PTEN/PI3K/Akt pathway and induced c-Myc and c-Jun. Knockdown of PTEN mimicked EBV-miR-BART7-3p-induced tumorigenic phenotype. Based on these results, we conducted a therapeutic experiment by using gold nano-particles (AuNPs) carrying anti-EBV-miR-BART7-3p. Silencing of EBV-miR-BART7-3p reduced tumor growth in animal model. We conclude that EBV-miR-BART7-3p favors carcinogenesis, representing a potential target for miRNA-based therapy.

No MeSH data available.


Related in: MedlinePlus