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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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Intravenous delivery of miR-99a-loaded NPs suppresses growth of HCC xenografts in nude mice.(A) Relative tumor volume of nude mice bearing HCC tumors treated with PBS, blank PEAL NPs, miR-99a-PEAL-LA NPs and miR-99a-PEAL-LA/VEGFab NPs. Data are presented as the means ± S.D. (n = 5). (B) Photographs of excised tumors from the different groups harvested at the endpoint of treatment. (C) Ki-67 analysis by immunohistochemistry and TUNEL assays of tumor tissues after treatment with the various formulations. The scale bar is 200 μm.
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f6: Intravenous delivery of miR-99a-loaded NPs suppresses growth of HCC xenografts in nude mice.(A) Relative tumor volume of nude mice bearing HCC tumors treated with PBS, blank PEAL NPs, miR-99a-PEAL-LA NPs and miR-99a-PEAL-LA/VEGFab NPs. Data are presented as the means ± S.D. (n = 5). (B) Photographs of excised tumors from the different groups harvested at the endpoint of treatment. (C) Ki-67 analysis by immunohistochemistry and TUNEL assays of tumor tissues after treatment with the various formulations. The scale bar is 200 μm.

Mentions: To explore the in vivo antitumor efficiency of NPs, we used a HepG2 subcutaneous xenograft nude mouse model to monitor tumor progression and test the superiority of the miR-99-PEAL-LA/VEGFab NPs delivery system. The NPs were injected through the tail vein, which is consistent with the anticipated clinical route of administration. As indicated in Fig. 6A and B, we observed no significant difference in tumor size among mice treated with PBS or blank PEAL NPs alone. In contrast, the tumor growth inhibition of mice treated with miR-99a-PEAL-LA NPs or miR-99a-PEAL-LA-VEGFab NPs was much higher than that of mice receiving PBS or PEAL NPs. The final tumor size of mice treated with miR-99a-PEAL-LA/VEGFab NPs was approximately 50 mm3, which was remarkably smaller than that of the other groups, confirming effective antitumor efficiency of this NP formulation in HCC-bearing mice. These results were further validated by immunohistochemistry and TUNEL assay analysis (Fig. 6C). The expression of Ki-67, a representative marker for proliferating cells, was detected to investigate the proliferative activity of HCC xenografts treated with NPs. The percentage of Ki-67-positive cells was remarkably decreased in the miR-99a-PEAL-LA/VEGFab group compared with the other groups (Fig. 6C). These data are consistent with the results of the TUNEL assay, in which apoptosis induced by miR-99a-PEAL-LA/VEGFab NPs treatment was increased compared with that of either PBS, PEAL NP or miR-99a-PEAL-LA NPs treatment (Fig. 6C). These results suggest that miR-99a-PEAL-LA/VEGFab NPs effectively suppress tumor growth by inhibiting proliferation and inducing apoptosis in HCC tumor cells.


PLGA-based dual targeted nanoparticles enhance miRNA transfection efficiency in hepatic carcinoma
Intravenous delivery of miR-99a-loaded NPs suppresses growth of HCC xenografts in nude mice.(A) Relative tumor volume of nude mice bearing HCC tumors treated with PBS, blank PEAL NPs, miR-99a-PEAL-LA NPs and miR-99a-PEAL-LA/VEGFab NPs. Data are presented as the means ± S.D. (n = 5). (B) Photographs of excised tumors from the different groups harvested at the endpoint of treatment. (C) Ki-67 analysis by immunohistochemistry and TUNEL assays of tumor tissues after treatment with the various formulations. The scale bar is 200 μm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5384185&req=5

f6: Intravenous delivery of miR-99a-loaded NPs suppresses growth of HCC xenografts in nude mice.(A) Relative tumor volume of nude mice bearing HCC tumors treated with PBS, blank PEAL NPs, miR-99a-PEAL-LA NPs and miR-99a-PEAL-LA/VEGFab NPs. Data are presented as the means ± S.D. (n = 5). (B) Photographs of excised tumors from the different groups harvested at the endpoint of treatment. (C) Ki-67 analysis by immunohistochemistry and TUNEL assays of tumor tissues after treatment with the various formulations. The scale bar is 200 μm.
Mentions: To explore the in vivo antitumor efficiency of NPs, we used a HepG2 subcutaneous xenograft nude mouse model to monitor tumor progression and test the superiority of the miR-99-PEAL-LA/VEGFab NPs delivery system. The NPs were injected through the tail vein, which is consistent with the anticipated clinical route of administration. As indicated in Fig. 6A and B, we observed no significant difference in tumor size among mice treated with PBS or blank PEAL NPs alone. In contrast, the tumor growth inhibition of mice treated with miR-99a-PEAL-LA NPs or miR-99a-PEAL-LA-VEGFab NPs was much higher than that of mice receiving PBS or PEAL NPs. The final tumor size of mice treated with miR-99a-PEAL-LA/VEGFab NPs was approximately 50 mm3, which was remarkably smaller than that of the other groups, confirming effective antitumor efficiency of this NP formulation in HCC-bearing mice. These results were further validated by immunohistochemistry and TUNEL assay analysis (Fig. 6C). The expression of Ki-67, a representative marker for proliferating cells, was detected to investigate the proliferative activity of HCC xenografts treated with NPs. The percentage of Ki-67-positive cells was remarkably decreased in the miR-99a-PEAL-LA/VEGFab group compared with the other groups (Fig. 6C). These data are consistent with the results of the TUNEL assay, in which apoptosis induced by miR-99a-PEAL-LA/VEGFab NPs treatment was increased compared with that of either PBS, PEAL NP or miR-99a-PEAL-LA NPs treatment (Fig. 6C). These results suggest that miR-99a-PEAL-LA/VEGFab NPs effectively suppress tumor growth by inhibiting proliferation and inducing apoptosis in HCC tumor cells.

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