Effects of RGD peptide-modified fenretinide liposomes on proliferation, apoptosis and migration of malignant melanoma cells / 中华皮肤科杂志
Chinese Journal of Dermatology
; (12): 182-188, 2019.
Article
in Zh
| WPRIM
| ID: wpr-745761
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WPRO
ABSTRACT
Objective To assess the effect of fenretinide (4-HPR) on proliferation,apoptosis and migration of B16F10 and A375 melanoma cells,and to evaluate the effect of liposomes and RGD peptidemodified liposomes on its uptake and therapeutic effects.Methods A film-hydration method was used to prepare 4-HPR liposomes (4-HPRL),which were modified with RGD peptide to prepare RGD-4-HPRL,and the concentration,particle size,electric potential,drug loading capacity and encapsulation efficiency were measured for 4-HPRL and RGD-4-HPRL.In vitro cultured B16F10 and A375 cells were divided into several groups4-HPR group,4-HPRL group and RGD-4-HPRL group treated with Dulbecco's minimum essential medium (DMEM) containing 4-HPR bulk drug,4-HPRL and RGD-4-HPRL respectively at the same concentration of 4-HPR,and control group treated with culture solution at the same volume.After different durations of treatment,cell counting kit-8 (CCK8) assay was performed to evaluate cellular proliferative activity,annexin V/propidium iodide staining to detect apoptosis,and scratch wound healing assay to evaluate the effect of drug treatment on cell migration ability.Then,4-HPR was replaced by coumarin 6 (C6) to prepare C6 liposomes (C6L) and RGD-C6L,and flow cytometry was conducted to evaluate C6 uptake by B16F10 cells.Statistical analysis was carried out with SPSS22.0 software by one-way analysis of variance (ANOVA) for the comparison among several groups and t test for the comparison between two groups.Results The concentration of 4-HPR in the prepared 4-HPRL solution was over 1 300 mg/L.The encapsulation efficiency and drug loading capacity of 4-HPRL were (95.51 ± 1.22)% and (7.27 ± 0.11)% respectively,and those of RGD-4-HPRL were (95.82 ± 0.81)% and (7.14 ± 0.13)% respectively.The particle size distribution of 4-HPRL and RGD-4-HPRL was uniform,and their average particle size was below 100 nm.CCK8 assay showed that 4-HPR could markedly inhibit the proliferative activities of B16F10 and A375 cells.The cell proliferation inhibition rate of 4-HPRL was higher than that of 4-HPR at the same concentration of 4-HPR (P < 0.01),and the inhibition rate of RGD-4-HPRL was higher than that of 4-HPRL (P < 0.01 or P < 0.05) and 4-HPR (P < 0.01).As annexin V/propidium iodide apoptosis assay showed,when the concentration of 4-HPR was 10 mg/L,the total apoptosis rates of B16F10 cells in the control group,4-HPR group,4-HPRL group and RGD-HPRL group were (4.44 ± 0.35)%,(28.33 ± 0.66)%,(46.43 ± 0.77)% and (51.33 ± 0.37)% respectively.When the concentration of 4-HPR was 20 mg/L,the total apoptosis rates of A375 cells in the above 4 groups were (4.97 ± 0.62)%,(16.68 ± 3.81)%,(32.62 ± 1.24)% and (44.85 ± 4.92)% respectively.The apoptosis rates of B16F10 and A375 cells were significantly higher in the 4-HPRL group than in the 4-HPR group (both P < 0.01),and higher in the RGD-4-HPRL group than in the 4-HPRL group (both P < 0.01) and 4-HPR group (both P <0.01).Scratch wound healing assay showed that 4-HPR could inhibit scratch healing and migration of B16F10 and A375 cells,and the inhibitory effects of 4-HPRL and RGD-4-HPRL were distinctly superior to those of 4-HPR bulk drug.C6 uptake assay revealed that the fluorescence intensity of C6 in B16F10 cells in the control group,C6 group,C6L group and RGD-C6L group were 2.15 ± 0.28,8.56 ± 0.36,20.48 ± 0.13 and 22.55 ± 0.07 respectively,and there were significant differences between the 4 groups (F =67 194.186,P < 0.01).Additionally,the fluorescence intensity of C6 was significantly higher in the C6L group and RGD-C6L group than in the C6 group (both P < 0.01),and higher in the RGD-C6L group than in the C6L Group (P < 0.01).Conclusions 4-HPR can inhibit the proliferation and migration of A375 and B16F10 cells,and induce their apoptosis.Liposomes and RGD-targeted liposomes can markedly enhance the effect of 4-HPR on melanoma cells.
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WPRIM
Language:
Zh
Journal:
Chinese Journal of Dermatology
Year:
2019
Type:
Article