Figures & data
Table 1 Particle size and zeta potential of liposomes at room temperature
Figure 1 Uptake of Tf-CL and CL in HepG2 and Hep3B cells.
Notes: (A) Fluorescence image of cellular uptake in HepG2. (B) Cellular uptake measured by flow cytometry. (C) Fluorescence image of cellular uptake in Hep3B. HepG2 cells and Hep3B cells were treated with different liposomes at 37°C for 4 hours. Block group was treated with Tf-CL and 100 µg/mL of free Tf. control group was untreated.
Abbreviations: Tf-CL, transferrin-targeted liposomal calcein; CL, nontargeted liposomal calcein; FI, fluorescence intensity; Tf, transferrin.
Figure 2 Uptake of Cy3-anti-miR-221-containing Tf-targeted liposome and nontargeted liposome in HepG2 cells was determined by fluorescence microscopy.
Notes: HepG2 cells were treated with different liposomes at 37°C for 4 hours. Control group was untreated. Block group was treated with Tf-RL plus 100 µg/mL of free Tf.
Abbreviations: RL, nontargeted liposome containing cy3-anti-miR-221; Tf-RL, transferrin-targeted liposome containing cy3-anti-miR-221; Tf, transferrin; DAPI, 4′,6-diamidino-2-phenylindole.
Figure 3 Cell viability of HepG2 cells.
Note: Each value represents the mean ± standard deviation (n=5)
Figure 4 Typical pictures of cellular apoptosis (A) and cell cycle (B) analyzed by flow cytometry.
Notes: HepG2 cells were treated with Tf-RL and RL for 24 hours. The final concentration of anti-miR-221 in these formulations was 200 nM. Control group was untreated.
Abbreviations: RL, nontargeted liposome containing anti-miR-221; Tf-RL, transferrin-targeted liposome containing anti-miR-221; FITC, fluorescein isothiocyanate; PI, propidium iodide.
Figure 5 In vitro silencing efficiency of Tf-RL and RL was determined by RT-PCR (A) and Western blotting (B) at gene and protein level, respectively.
Notes: HepG2 cells were treated with Tf-RL and RL for 24 hours for RT-PCR assay and 48 hours for Western blotting. The final concentration of anti-miR-221 in these formulations was 200 nM. Control group was untreated. Each value represents the mean ± standard deviation (n=3). *P<0.05.
Abbreviations: RL, nontargeted liposome containing anti-miR-221; Tf-RL, transferrin-targeted liposome containing anti-miR-221; RT-PCR, real-time polymerase chain reaction.
Figure 6 Tissue distribution of Cy3-anti-miR-221 liposome in normal Kunming mice.
Notes: The heart, lung, spleen, kidney, and liver were harvested from female Kunming mice 4 hours after intravenous administration of Cy3-anti-miR-221-containing nontargeted liposome or transferrin-targeted liposome. Cy3 fluorescence signals were measured by IVIs imaging. Control group was treated with saline.
Abbreviations: RL, nontargeted liposome containing anti-miR-221; Tf-RL, transferrin-targeted liposome containing anti-miR-221.
Figure 7 Tissue distribution of Cy3-anti-miR-221 liposome in a xenograft model of HepG2 human liver cancer in female BALB/c-nu mice.
Notes: Tumors from the heart, lung, spleen, kidney, and liver were harvested from female BALB/c-nu mice 4 hours after intravenous administration of Cy3-anti-miR-221-containing nontargeted liposome or transferrin-targeted liposome. Cy3 fluorescence signals were measured by IVIS imaging. Control group was treated with saline.
Abbreviations: RL, nontargeted liposome containing anti-miR-221; Tf-RL, transferrin-targeted liposome containing anti-miR-221.
Figure 8 In vivo evaluation of the effect of anti-miR-221 treatments on target gene expressions in a xenograft model of HepG2 human liver cancer.
Notes: Female BALB/c-nu mice were treated with 1.2 mg/kg of Tf-RL and RL for 48 hours. Tumors were harvested, and the target gene expressions were determined by RT-PCR. Control group was treated with saline. Each value represents the mean ± standard deviation (n=3). *P<0.05.
Abbreviations: RL, nontargeted liposome containing anti-miR-221; Tf-RL, transferrin-targeted liposome containing anti-miR-221; RT-PCR, real-time polymerase chain reaction.
