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PLGA-based dual targeted nanoparticles enhance miRNA transfection efficiency in hepatic carcinoma

View Article: PubMed Central - PubMed

ABSTRACT

Hepatic carcinoma (HCC) is a lethal disease associated with high morbidity and poor prognosis. Recently years, gene therapies have offered novel modalities to improve the prognosis of HCC patients. MicroRNA-99a (miR-99a) is frequently down-regulated in HCC, where it acts as a tumor suppressor. Therefore, we constructed monomethoxy (polyethylene glycol)-poly(D,L-lactide-co-glycolide)-poly(L-lysine)-lactobionic acid- anti-vascular endothelial growth factor antibody (mPEG-PLGA-PLL-LA/VEGFab or PEAL-LA/VEGFab) nanoparticles (NPs) with highly specific targeting properties as carriers to restore the expression of miR-99a both in vitro and in vivo, to inhibit HCC progression. In vitro, PEAL-LA/VEGFab NPs showed more efficient delivery of miR-99a to HepG2 cells than the conventional transfection reagent LipofectamineTM2000 (Lip2000). The higher delivery efficiency associated with PEAL-LA/VEGFab NPs consequently resulted in down-regulation of target genes and suppression of the proliferation, migration and invasion of HepG2 cells. In vivo, miR-99a-PEAL-LA/VEGFab NPs inhibited tumor xenograft growth in HCC-bearing mice without causing obvious systemic toxicity. Our results demonstrate that PEAL-LA/VEGFab NPs selectively and effectively deliver miR-99a to HCC cells based on the double-targeting character of these nanoparticles, thereby offering potential for translation into effective clinical therapies for HCC.

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(A) The effects on colony formation of HepG2 cells treated with PBS (a), miR-99a-Lip2000 (b), miR-99a-PEAL-LA NPs (c) and miR-99a-PEAL-LA-VEGFab NPs (d) was evaluated by the clonogenic assay. (B) The corresponding quantification of colony formation efficiency. Data are presented as the means ± S.D., (n = 5). **p < 0.01. (C) Inhibition of migration and invasion of HepG2 cells upon treatment with PBS, miR-99a-Lip2000, miR-99a-PEAL-LA NPs and miR-99a-PEAL-LA/VEGFab NPs. Data are represented as the means ± S.D. (N = 3). Magnification 100 ×, the scale bar is 200 μm. (D) The corresponding quantification of migration and invasion. (n = 3) **p < 0.01.
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f5: (A) The effects on colony formation of HepG2 cells treated with PBS (a), miR-99a-Lip2000 (b), miR-99a-PEAL-LA NPs (c) and miR-99a-PEAL-LA-VEGFab NPs (d) was evaluated by the clonogenic assay. (B) The corresponding quantification of colony formation efficiency. Data are presented as the means ± S.D., (n = 5). **p < 0.01. (C) Inhibition of migration and invasion of HepG2 cells upon treatment with PBS, miR-99a-Lip2000, miR-99a-PEAL-LA NPs and miR-99a-PEAL-LA/VEGFab NPs. Data are represented as the means ± S.D. (N = 3). Magnification 100 ×, the scale bar is 200 μm. (D) The corresponding quantification of migration and invasion. (n = 3) **p < 0.01.

Mentions: Next, we evaluated the ability of NPs to inhibit tumor progression in vitro. First, the clonogenic assay was performed to investigate the inhibitory effect of the NPs on clonogenic formation in HepG2 cells. As shown in Fig. 5A and B, a reduced number of colonies was observed in cells treated with miR-99a-Lip2000 and miR-99a-PEAL-LA NPs compared with the control group (38.40 ± 1.36 vs. 77.60 ± 1.44, P < 0.01; 36.00 ± 2.43 vs. 77.60 ± 1.44, P < 0.01). Moreover, more effective inhibition was observed in cells treated with miR-99a-PEAL-LA/VEGFab NPs (24.00 ± 1.00 vs. 77.60 ± 1.43, P < 0.01), which exhibited an obviously higher inhibition rate of 69.1% for clonogenic growth potential in HCC. These results suggest that the dual-targeting PEAL-LA/VEGFab NPs could deliver miR-99a into HepG2 cells most effectively, which results in the inhibition of the proliferation of tumor cells and enhanced anti-tumor activity in vitro.


PLGA-based dual targeted nanoparticles enhance miRNA transfection efficiency in hepatic carcinoma
(A) The effects on colony formation of HepG2 cells treated with PBS (a), miR-99a-Lip2000 (b), miR-99a-PEAL-LA NPs (c) and miR-99a-PEAL-LA-VEGFab NPs (d) was evaluated by the clonogenic assay. (B) The corresponding quantification of colony formation efficiency. Data are presented as the means ± S.D., (n = 5). **p < 0.01. (C) Inhibition of migration and invasion of HepG2 cells upon treatment with PBS, miR-99a-Lip2000, miR-99a-PEAL-LA NPs and miR-99a-PEAL-LA/VEGFab NPs. Data are represented as the means ± S.D. (N = 3). Magnification 100 ×, the scale bar is 200 μm. (D) The corresponding quantification of migration and invasion. (n = 3) **p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5384185&req=5

f5: (A) The effects on colony formation of HepG2 cells treated with PBS (a), miR-99a-Lip2000 (b), miR-99a-PEAL-LA NPs (c) and miR-99a-PEAL-LA-VEGFab NPs (d) was evaluated by the clonogenic assay. (B) The corresponding quantification of colony formation efficiency. Data are presented as the means ± S.D., (n = 5). **p < 0.01. (C) Inhibition of migration and invasion of HepG2 cells upon treatment with PBS, miR-99a-Lip2000, miR-99a-PEAL-LA NPs and miR-99a-PEAL-LA/VEGFab NPs. Data are represented as the means ± S.D. (N = 3). Magnification 100 ×, the scale bar is 200 μm. (D) The corresponding quantification of migration and invasion. (n = 3) **p < 0.01.
Mentions: Next, we evaluated the ability of NPs to inhibit tumor progression in vitro. First, the clonogenic assay was performed to investigate the inhibitory effect of the NPs on clonogenic formation in HepG2 cells. As shown in Fig. 5A and B, a reduced number of colonies was observed in cells treated with miR-99a-Lip2000 and miR-99a-PEAL-LA NPs compared with the control group (38.40 ± 1.36 vs. 77.60 ± 1.44, P < 0.01; 36.00 ± 2.43 vs. 77.60 ± 1.44, P < 0.01). Moreover, more effective inhibition was observed in cells treated with miR-99a-PEAL-LA/VEGFab NPs (24.00 ± 1.00 vs. 77.60 ± 1.43, P < 0.01), which exhibited an obviously higher inhibition rate of 69.1% for clonogenic growth potential in HCC. These results suggest that the dual-targeting PEAL-LA/VEGFab NPs could deliver miR-99a into HepG2 cells most effectively, which results in the inhibition of the proliferation of tumor cells and enhanced anti-tumor activity in vitro.

View Article: PubMed Central - PubMed

ABSTRACT

Hepatic carcinoma (HCC) is a lethal disease associated with high morbidity and poor prognosis. Recently years, gene therapies have offered novel modalities to improve the prognosis of HCC patients. MicroRNA-99a (miR-99a) is frequently down-regulated in HCC, where it acts as a tumor suppressor. Therefore, we constructed monomethoxy (polyethylene glycol)-poly(D,L-lactide-co-glycolide)-poly(L-lysine)-lactobionic acid- anti-vascular endothelial growth factor antibody (mPEG-PLGA-PLL-LA/VEGFab or PEAL-LA/VEGFab) nanoparticles (NPs) with highly specific targeting properties as carriers to restore the expression of miR-99a both in vitro and in vivo, to inhibit HCC progression. In vitro, PEAL-LA/VEGFab NPs showed more efficient delivery of miR-99a to HepG2 cells than the conventional transfection reagent LipofectamineTM2000 (Lip2000). The higher delivery efficiency associated with PEAL-LA/VEGFab NPs consequently resulted in down-regulation of target genes and suppression of the proliferation, migration and invasion of HepG2 cells. In vivo, miR-99a-PEAL-LA/VEGFab NPs inhibited tumor xenograft growth in HCC-bearing mice without causing obvious systemic toxicity. Our results demonstrate that PEAL-LA/VEGFab NPs selectively and effectively deliver miR-99a to HCC cells based on the double-targeting character of these nanoparticles, thereby offering potential for translation into effective clinical therapies for HCC.

No MeSH data available.


Related in: MedlinePlus