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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.

No MeSH data available.


Related in: MedlinePlus

Caveolae-mediated cellular internalization and intracellular trafficking of CNCs in 4T1 cells.Colocalization of (a) C6CNCs or (b) CNCsCy5-RNA with caveolae after incubation for 4 h at 37 °C in the absence or presence of nystatin. Caveolin-1 (CAV-1) was marked by anti-caveolin-1 antibody/Alexa Fluors® 594 (red) or 488 (green). Colocalization of (c) C6CNCs or (d) CNCsCy5-RNA with CTB-Alexa Fluors® 594 (red) or 488 (green) after incubation for 4 h at 37 °C in the absence or presence of nystatin. Yellow spots indicate the colocalization of CNCs with caveolae or CTB. 4T1 cells without treatment were used as the control. Scale bar: 10 μm. (e) Colocalization of CNCs with lysosomes after incubation for 4 h at 37 °C. Scale bar: 10 μm. (f) Cytosolic location of the CNCs in 4T1 cells observed by TEM after incubation for 4 h at 37 °C. Control group was the normal cells without treatment. Red arrow heads indicate CNCs. Cyt, cytoplasm; N, nucleus; V, vesicle; L: lysosome. Scale bar: 0.5 μm.
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f5: Caveolae-mediated cellular internalization and intracellular trafficking of CNCs in 4T1 cells.Colocalization of (a) C6CNCs or (b) CNCsCy5-RNA with caveolae after incubation for 4 h at 37 °C in the absence or presence of nystatin. Caveolin-1 (CAV-1) was marked by anti-caveolin-1 antibody/Alexa Fluors® 594 (red) or 488 (green). Colocalization of (c) C6CNCs or (d) CNCsCy5-RNA with CTB-Alexa Fluors® 594 (red) or 488 (green) after incubation for 4 h at 37 °C in the absence or presence of nystatin. Yellow spots indicate the colocalization of CNCs with caveolae or CTB. 4T1 cells without treatment were used as the control. Scale bar: 10 μm. (e) Colocalization of CNCs with lysosomes after incubation for 4 h at 37 °C. Scale bar: 10 μm. (f) Cytosolic location of the CNCs in 4T1 cells observed by TEM after incubation for 4 h at 37 °C. Control group was the normal cells without treatment. Red arrow heads indicate CNCs. Cyt, cytoplasm; N, nucleus; V, vesicle; L: lysosome. Scale bar: 0.5 μm.

Mentions: The caveolae-mediated cellular internalization of CNCs in 4T1 and Caco-2 cells was further demonstrated using anti-caveolin-1 antibody-Caveolae Marker (Alexa Fluors® 488 or 594) and the pathway marker of caveolae-mediated internalization, cholera toxin subunit B (CTB)-Alexa Fluor® 488 (or 594). Figure 5a and b shows the colocalization of CNCs with caveolae in 4T1 cells. The yellow areas are from the merged images, indicating the high colocalization of the CNCs with caveolae. In contrast, the cellular internalization of CNCs decreased significantly in the presence of nystatin, an inhibitor that can deplete cholesterol from membranes and block the formation of caveolae. Figure 5c and d shows the high colocalization of CNCs with the pathway marker of caveolae-mediated internalization, CTB. The addition of nystatin significantly inhibited the cellular uptake of the CNCs and CTB. The caveolae-mediated cellular internalization of CNCs was further confirmed in Caco-2 cells, intestinal epithelial cells possessing abundant caveolaes27. As shown in Supplementary Fig. S7, similar results were observed in Caco-2 cells. These results suggested that the CNCs could be internalized via the caveolae-mediated pathway.


Cytosolic co-delivery of miRNA-34a and docetaxel with core-shell nanocarriers via caveolae-mediated pathway for the treatment of metastatic breast cancer
Caveolae-mediated cellular internalization and intracellular trafficking of CNCs in 4T1 cells.Colocalization of (a) C6CNCs or (b) CNCsCy5-RNA with caveolae after incubation for 4 h at 37 °C in the absence or presence of nystatin. Caveolin-1 (CAV-1) was marked by anti-caveolin-1 antibody/Alexa Fluors® 594 (red) or 488 (green). Colocalization of (c) C6CNCs or (d) CNCsCy5-RNA with CTB-Alexa Fluors® 594 (red) or 488 (green) after incubation for 4 h at 37 °C in the absence or presence of nystatin. Yellow spots indicate the colocalization of CNCs with caveolae or CTB. 4T1 cells without treatment were used as the control. Scale bar: 10 μm. (e) Colocalization of CNCs with lysosomes after incubation for 4 h at 37 °C. Scale bar: 10 μm. (f) Cytosolic location of the CNCs in 4T1 cells observed by TEM after incubation for 4 h at 37 °C. Control group was the normal cells without treatment. Red arrow heads indicate CNCs. Cyt, cytoplasm; N, nucleus; V, vesicle; L: lysosome. Scale bar: 0.5 μm.
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f5: Caveolae-mediated cellular internalization and intracellular trafficking of CNCs in 4T1 cells.Colocalization of (a) C6CNCs or (b) CNCsCy5-RNA with caveolae after incubation for 4 h at 37 °C in the absence or presence of nystatin. Caveolin-1 (CAV-1) was marked by anti-caveolin-1 antibody/Alexa Fluors® 594 (red) or 488 (green). Colocalization of (c) C6CNCs or (d) CNCsCy5-RNA with CTB-Alexa Fluors® 594 (red) or 488 (green) after incubation for 4 h at 37 °C in the absence or presence of nystatin. Yellow spots indicate the colocalization of CNCs with caveolae or CTB. 4T1 cells without treatment were used as the control. Scale bar: 10 μm. (e) Colocalization of CNCs with lysosomes after incubation for 4 h at 37 °C. Scale bar: 10 μm. (f) Cytosolic location of the CNCs in 4T1 cells observed by TEM after incubation for 4 h at 37 °C. Control group was the normal cells without treatment. Red arrow heads indicate CNCs. Cyt, cytoplasm; N, nucleus; V, vesicle; L: lysosome. Scale bar: 0.5 μm.
Mentions: The caveolae-mediated cellular internalization of CNCs in 4T1 and Caco-2 cells was further demonstrated using anti-caveolin-1 antibody-Caveolae Marker (Alexa Fluors® 488 or 594) and the pathway marker of caveolae-mediated internalization, cholera toxin subunit B (CTB)-Alexa Fluor® 488 (or 594). Figure 5a and b shows the colocalization of CNCs with caveolae in 4T1 cells. The yellow areas are from the merged images, indicating the high colocalization of the CNCs with caveolae. In contrast, the cellular internalization of CNCs decreased significantly in the presence of nystatin, an inhibitor that can deplete cholesterol from membranes and block the formation of caveolae. Figure 5c and d shows the high colocalization of CNCs with the pathway marker of caveolae-mediated internalization, CTB. The addition of nystatin significantly inhibited the cellular uptake of the CNCs and CTB. The caveolae-mediated cellular internalization of CNCs was further confirmed in Caco-2 cells, intestinal epithelial cells possessing abundant caveolaes27. As shown in Supplementary Fig. S7, similar results were observed in Caco-2 cells. These results suggested that the CNCs could be internalized via the caveolae-mediated pathway.

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.

No MeSH data available.


Related in: MedlinePlus