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Cytosolic co-delivery of miRNA-34a and docetaxel with core-shell nanocarriers via caveolae-mediated pathway for the treatment of metastatic breast cancer

View Article: PubMed Central - PubMed

ABSTRACT

-: Codelivery of microRNAs and chemotherapeutic drugs into tumor cells is an attractive strategy for synergetic breast cancer therapy due to their complementary mechanisms. In this work, a core-shell nanocarrier coated by cationic albumin was developed to simultaneously deliver miRNA-34a and docetaxel (DTX) into breast cancer cells for improved therapeutic effect. The co-delivery nanocarriers showed a spherical morphology with an average particle size of 183.9 nm, and they efficiently protected miRNA-34a from degradation by RNase and serum. Importantly, the nanocarriers entered the cytosol via a caveolae-mediated pathway without entrapment in endosomes/lysosomes, thus improving the utilization of the cargo. In vitro, the co-delivery nanocarriers suppressed the expression of anti-apoptosis gene Bcl-2 at both transcription and protein levels, inhibited tumor cell migration and efficiently induced cell apoptosis and cytotoxicity. In vivo, the co-delivery nanocarriers prolonged the blood circulation of DTX, enhanced tumor accumulation of the cargo and significantly inhibited tumor growth and metastasis in 4T1-tumor bearing mice models. Taken together, the present nanocarrier co-loading with DTX and miRNA-34a is a new nanoplatform for the combination of insoluble drugs and gene/protein drugs and provides a promising strategy for the treatment of metastatic breast cancer.

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In vitro antitumor efficacy.(a) Cell cytotoxicity of 4T1 cells and A549 cells treated with BNCs of different concentrations at 37 °C. (b) Cell cytotoxicity of 4T1 cells treated with different formulations at 37 °C. The concentration of miRNA-34a was fixed at 100 nM. *P < 0.05, **P < 0.01, ***P < 0.001. (c) Cell apoptosis induction and morphological changes in 4T1 cells treated with different formulations imaged by fluorescence microscopy. Cell nuclei were stained with DAPI. The concentrations of miRNA-34a and DTX were 100 nM and 10 μg/mL, respectively. The control group was the normal cells without treatment. Red arrow heads indicate the typical apoptosis features such as chromatin condensation, fragmentation and apoptotic body formation. Scale bar: 50 μm. (d) Cell apoptosis detected by FCM using an AnnexinV-FITC/PI apoptosis detection kit. (e) Inhibition migration of 4T1 cells examined by transwell chamber assay at 37 °C. Blue regions indicate the migrated cells. The control group was the normal cells without treatment. Scale bar: 50 μm.
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f7: In vitro antitumor efficacy.(a) Cell cytotoxicity of 4T1 cells and A549 cells treated with BNCs of different concentrations at 37 °C. (b) Cell cytotoxicity of 4T1 cells treated with different formulations at 37 °C. The concentration of miRNA-34a was fixed at 100 nM. *P < 0.05, **P < 0.01, ***P < 0.001. (c) Cell apoptosis induction and morphological changes in 4T1 cells treated with different formulations imaged by fluorescence microscopy. Cell nuclei were stained with DAPI. The concentrations of miRNA-34a and DTX were 100 nM and 10 μg/mL, respectively. The control group was the normal cells without treatment. Red arrow heads indicate the typical apoptosis features such as chromatin condensation, fragmentation and apoptotic body formation. Scale bar: 50 μm. (d) Cell apoptosis detected by FCM using an AnnexinV-FITC/PI apoptosis detection kit. (e) Inhibition migration of 4T1 cells examined by transwell chamber assay at 37 °C. Blue regions indicate the migrated cells. The control group was the normal cells without treatment. Scale bar: 50 μm.

Mentions: The in vitro cytotoxicity of CNCs was evaluated by MTT assay. As shown in Fig. 7a, the viability of A549 and 4T1 cells treated with BNCs was higher than 80% as the concentration of CBSA changes from 0.1 to 500 μg/mL, indicating that the blank nanocarriers had low cytotoxicity under the tested concentrations. Free DTX, DNCs and CNCs exhibited concentration dependent cytotoxicity on 4T1 cells (Fig. 7b). DNCs showed slightly higher cytotoxicity than free DTX at the same drug concentration, which could be attributed to the higher uptake efficiency of drugs by nanocarriers. The cells treated with RNCs displayed a reduced viability at a fixed miRNA-34a concentration of 100 nM. Although miRNA-34a can inhibit cell proliferation, it cannot induce strong cytotoxicity completely. However, compared with free DTX, DNCs or RNCs at the same dose, significant decreases of cell viability were observed in the cells treated with the CNCs, suggesting the synergistic cytotoxicity by co-delivery of DTX and miRNA-34a.


Cytosolic co-delivery of miRNA-34a and docetaxel with core-shell nanocarriers via caveolae-mediated pathway for the treatment of metastatic breast cancer
In vitro antitumor efficacy.(a) Cell cytotoxicity of 4T1 cells and A549 cells treated with BNCs of different concentrations at 37 °C. (b) Cell cytotoxicity of 4T1 cells treated with different formulations at 37 °C. The concentration of miRNA-34a was fixed at 100 nM. *P < 0.05, **P < 0.01, ***P < 0.001. (c) Cell apoptosis induction and morphological changes in 4T1 cells treated with different formulations imaged by fluorescence microscopy. Cell nuclei were stained with DAPI. The concentrations of miRNA-34a and DTX were 100 nM and 10 μg/mL, respectively. The control group was the normal cells without treatment. Red arrow heads indicate the typical apoptosis features such as chromatin condensation, fragmentation and apoptotic body formation. Scale bar: 50 μm. (d) Cell apoptosis detected by FCM using an AnnexinV-FITC/PI apoptosis detection kit. (e) Inhibition migration of 4T1 cells examined by transwell chamber assay at 37 °C. Blue regions indicate the migrated cells. The control group was the normal cells without treatment. Scale bar: 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5382875&req=5

f7: In vitro antitumor efficacy.(a) Cell cytotoxicity of 4T1 cells and A549 cells treated with BNCs of different concentrations at 37 °C. (b) Cell cytotoxicity of 4T1 cells treated with different formulations at 37 °C. The concentration of miRNA-34a was fixed at 100 nM. *P < 0.05, **P < 0.01, ***P < 0.001. (c) Cell apoptosis induction and morphological changes in 4T1 cells treated with different formulations imaged by fluorescence microscopy. Cell nuclei were stained with DAPI. The concentrations of miRNA-34a and DTX were 100 nM and 10 μg/mL, respectively. The control group was the normal cells without treatment. Red arrow heads indicate the typical apoptosis features such as chromatin condensation, fragmentation and apoptotic body formation. Scale bar: 50 μm. (d) Cell apoptosis detected by FCM using an AnnexinV-FITC/PI apoptosis detection kit. (e) Inhibition migration of 4T1 cells examined by transwell chamber assay at 37 °C. Blue regions indicate the migrated cells. The control group was the normal cells without treatment. Scale bar: 50 μm.
Mentions: The in vitro cytotoxicity of CNCs was evaluated by MTT assay. As shown in Fig. 7a, the viability of A549 and 4T1 cells treated with BNCs was higher than 80% as the concentration of CBSA changes from 0.1 to 500 μg/mL, indicating that the blank nanocarriers had low cytotoxicity under the tested concentrations. Free DTX, DNCs and CNCs exhibited concentration dependent cytotoxicity on 4T1 cells (Fig. 7b). DNCs showed slightly higher cytotoxicity than free DTX at the same drug concentration, which could be attributed to the higher uptake efficiency of drugs by nanocarriers. The cells treated with RNCs displayed a reduced viability at a fixed miRNA-34a concentration of 100 nM. Although miRNA-34a can inhibit cell proliferation, it cannot induce strong cytotoxicity completely. However, compared with free DTX, DNCs or RNCs at the same dose, significant decreases of cell viability were observed in the cells treated with the CNCs, suggesting the synergistic cytotoxicity by co-delivery of DTX and miRNA-34a.

View Article: PubMed Central - PubMed

ABSTRACT

-: Codelivery of microRNAs and chemotherapeutic drugs into tumor cells is an attractive strategy for synergetic breast cancer therapy due to their complementary mechanisms. In this work, a core-shell nanocarrier coated by cationic albumin was developed to simultaneously deliver miRNA-34a and docetaxel (DTX) into breast cancer cells for improved therapeutic effect. The co-delivery nanocarriers showed a spherical morphology with an average particle size of 183.9&thinsp;nm, and they efficiently protected miRNA-34a from degradation by RNase and serum. Importantly, the nanocarriers entered the cytosol via a caveolae-mediated pathway without entrapment in endosomes/lysosomes, thus improving the utilization of the cargo. In vitro, the co-delivery nanocarriers suppressed the expression of anti-apoptosis gene Bcl-2 at both transcription and protein levels, inhibited tumor cell migration and efficiently induced cell apoptosis and cytotoxicity. In vivo, the co-delivery nanocarriers prolonged the blood circulation of DTX, enhanced tumor accumulation of the cargo and significantly inhibited tumor growth and metastasis in 4T1-tumor bearing mice models. Taken together, the present nanocarrier co-loading with DTX and miRNA-34a is a new nanoplatform for the combination of insoluble drugs and gene/protein drugs and provides a promising strategy for the treatment of metastatic breast cancer.

No MeSH data available.


Related in: MedlinePlus