Limits...
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

Silencing of EBV-miR-BART7-3p reduced the in vitro growth of EBV-positive NPC cells(A) A layer-by-layer method was applied to prepare gold-PEI nanoparticles carrying anti-miRNA. (B) The transfection efficiency of nanoparticles carrying anti-miRNA was evaluated by confocal microscopy at cellular level. (C) The inhibition efficiency of nano-anti-miR in HONE1-EBV cells was evaluated by qPCR in different time points. Data were shown as the mean ± SEM (**P < 0.01, ***P < 0.001). (D) Cell growth ability was evaluated by MTT assay in HONE1-EBV cells after transfected with anti-miR or anti-C. Data were shown as the mean ± SEM (***P < 0.001). (E) The expression levels of PTEN, p-Akt, c-Myc and CCND1 were analyzed by western blot in HONE1/EBV cells after transfected with anti-miR or anti-C. β-actin served as an internal control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4480720&req=5

Figure 4: Silencing of EBV-miR-BART7-3p reduced the in vitro growth of EBV-positive NPC cells(A) A layer-by-layer method was applied to prepare gold-PEI nanoparticles carrying anti-miRNA. (B) The transfection efficiency of nanoparticles carrying anti-miRNA was evaluated by confocal microscopy at cellular level. (C) The inhibition efficiency of nano-anti-miR in HONE1-EBV cells was evaluated by qPCR in different time points. Data were shown as the mean ± SEM (**P < 0.01, ***P < 0.001). (D) Cell growth ability was evaluated by MTT assay in HONE1-EBV cells after transfected with anti-miR or anti-C. Data were shown as the mean ± SEM (***P < 0.001). (E) The expression levels of PTEN, p-Akt, c-Myc and CCND1 were analyzed by western blot in HONE1/EBV cells after transfected with anti-miR or anti-C. β-actin served as an internal control.

Mentions: Given that EBV-miR-BART7-3p modulated growth promotion, we further explored whether or not EBV-miR-BART7-3p was a potential therapeutic target for NPC. We firstly analyzed EBV-miR-BART7-3p expression by qPCR in several EBV-positive NPC cell lines. As shown in Figure S7, there was a similar expression level of EBV-miR-BART7-3p in EBV positive NPC cells to the pooled NPC tissues. Of these EBV-positive NPC cell lines, HONE1-EBV cell line was selected as a representative cellular model because it grew better and was suitable for the following in vitro and in vivo experiments [24]. Subsequently, we fabricated a nano-carrier (gold-PEI) using a layer-by-layer method [30] (Figure 4A, see materials and methods) to deliver anti-EBV-miR-BART7-3p into HONE1-EBV cells. The size of Gold-PEI nano-carrier was about 20–30 nm as measured by dynamic lighting scatter (DLS) (Figure S8A). The zeta potential of gold-PEI was positive charge (about 20 mV) that facilitated the absorption of anti-miR with negative charge (Figure S8B). The nano-anti-miR displayed an approximately spherical shape with good dispersion and its size was similar to that of gold-PEI observed by TEM (Figure S9). To validate the transfection efficiency at the cellular level, we next investigated the cellular uptake of nano-carrier with FAM-anti-miR using confocal microscopy. There were much more visible green fluorescence particles in HONE-EBV cells transfected with nano-anti-miR (Figure 4B). Furthermore, the inhibition efficiency of nano-anti-miR was evaluated. qPCR revealed that nano-anti-miR effectively inhibited EBV-miR-BART-3p expression level in HONE-EBV cells in the first three days (Figure 4C). After treating with nano-anti-miR, cell growth was accordingly reduced (Figure 4D) and PTEN expression was obviously increased in HONE-EBV while the relevant signals (p-Akt, c-Myc and CCND1) were decreased (Figure 4E).


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)

Silencing of EBV-miR-BART7-3p reduced the in vitro growth of EBV-positive NPC cells(A) A layer-by-layer method was applied to prepare gold-PEI nanoparticles carrying anti-miRNA. (B) The transfection efficiency of nanoparticles carrying anti-miRNA was evaluated by confocal microscopy at cellular level. (C) The inhibition efficiency of nano-anti-miR in HONE1-EBV cells was evaluated by qPCR in different time points. Data were shown as the mean ± SEM (**P < 0.01, ***P < 0.001). (D) Cell growth ability was evaluated by MTT assay in HONE1-EBV cells after transfected with anti-miR or anti-C. Data were shown as the mean ± SEM (***P < 0.001). (E) The expression levels of PTEN, p-Akt, c-Myc and CCND1 were analyzed by western blot in HONE1/EBV cells after transfected with anti-miR or anti-C. β-actin served as an internal control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Silencing of EBV-miR-BART7-3p reduced the in vitro growth of EBV-positive NPC cells(A) A layer-by-layer method was applied to prepare gold-PEI nanoparticles carrying anti-miRNA. (B) The transfection efficiency of nanoparticles carrying anti-miRNA was evaluated by confocal microscopy at cellular level. (C) The inhibition efficiency of nano-anti-miR in HONE1-EBV cells was evaluated by qPCR in different time points. Data were shown as the mean ± SEM (**P < 0.01, ***P < 0.001). (D) Cell growth ability was evaluated by MTT assay in HONE1-EBV cells after transfected with anti-miR or anti-C. Data were shown as the mean ± SEM (***P < 0.001). (E) The expression levels of PTEN, p-Akt, c-Myc and CCND1 were analyzed by western blot in HONE1/EBV cells after transfected with anti-miR or anti-C. β-actin served as an internal control.
Mentions: Given that EBV-miR-BART7-3p modulated growth promotion, we further explored whether or not EBV-miR-BART7-3p was a potential therapeutic target for NPC. We firstly analyzed EBV-miR-BART7-3p expression by qPCR in several EBV-positive NPC cell lines. As shown in Figure S7, there was a similar expression level of EBV-miR-BART7-3p in EBV positive NPC cells to the pooled NPC tissues. Of these EBV-positive NPC cell lines, HONE1-EBV cell line was selected as a representative cellular model because it grew better and was suitable for the following in vitro and in vivo experiments [24]. Subsequently, we fabricated a nano-carrier (gold-PEI) using a layer-by-layer method [30] (Figure 4A, see materials and methods) to deliver anti-EBV-miR-BART7-3p into HONE1-EBV cells. The size of Gold-PEI nano-carrier was about 20–30 nm as measured by dynamic lighting scatter (DLS) (Figure S8A). The zeta potential of gold-PEI was positive charge (about 20 mV) that facilitated the absorption of anti-miR with negative charge (Figure S8B). The nano-anti-miR displayed an approximately spherical shape with good dispersion and its size was similar to that of gold-PEI observed by TEM (Figure S9). To validate the transfection efficiency at the cellular level, we next investigated the cellular uptake of nano-carrier with FAM-anti-miR using confocal microscopy. There were much more visible green fluorescence particles in HONE-EBV cells transfected with nano-anti-miR (Figure 4B). Furthermore, the inhibition efficiency of nano-anti-miR was evaluated. qPCR revealed that nano-anti-miR effectively inhibited EBV-miR-BART-3p expression level in HONE-EBV cells in the first three days (Figure 4C). After treating with nano-anti-miR, cell growth was accordingly reduced (Figure 4D) and PTEN expression was obviously increased in HONE-EBV while the relevant signals (p-Akt, c-Myc and CCND1) were decreased (Figure 4E).

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