ABSTRACT
Objective:To investigate the effect of propofol on proliferation, invasion and migration of human melanoma cell line A375 via miR-137/fibroblast growth factor 9 (FGF9) pathway.Methods:A375 cells were cultured in vitro. The half inhibitory concentration and half inhibitory time of propofol on A375 cells were determined by methyl thiazolyl tetrazolium (MTT) method. miR-137 mimics and miR-137 inhibitors were transfected into A375 cells by lipofectamine method. A375 cells were divided into control group, propofol group, miR-137 mimics group, propofol+ miR-137 mimics group, miR-137 inhibitors group and propofol+ miR-137 inhibitors group. After treated with the optimal time and concentration, the mRNA expression of miR-137 and FGF9 was detected by real time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR), and the invasion and migration ability of cells in vitro was detected by transwell method. Results:With the increase of propofol concentration, the proliferation rate of A375 cells was gradually decreased, and 80 μmol/L was selected as the half inhibition concentration. With the increase of propofol action time, the proliferation rate of A375 cells was gradually decreased, and 48 hours was selected as the half inhibition time. Compared with the control group, propofol could promote the expression of miR-137 mRNA, inhibit the expression of FGF9 mRNA, and inhibit the invasion and migration of A375 cells ( P<0.05). miR-137 mimics could promote the expression of miR-137 mRNA, inhibit the expression of FGF9 mRNA, and inhibit the invasion and migration of A375 cells. At the same time, after propofol intervention, the effect of promoting the expression of miR-137 mRNA, inhibiting the expression of FGF9 mRNA and inhibiting the invasion and migration of A375 cells was more significant ( P<0.05); miR-137 inhibitors could inhibit the expression of miR-137 mRNA, promote the expression of FGF9 mRNA, and promote the invasion and migration of A375 cells ( P<0.05). At the same time, after propofol intervention, the effects of inhibiting the expression of miR-137 mRNA, promoting the expression of FGF9 mRNA and promoting the invasion and migration of A375 cells were inhibited ( P<0.05). Conclusions:Propofol can inhibit the proliferation, invasion and migration of human melanoma cell line A375. The mechanism may be related to the inhibition of miR-137/FGF9 pathway activation by propofol.
ABSTRACT
The expression of fibroblast growth factor 9 (FGF9) recombinant fusion protein in Carthamus tinctorius was used to identify its effect on hair regrowth and wound repair system in mice, providing a basis for C. tinctorius as a plant bioreactor, and establishing a foundation for commercial applications of FGF9 fusion protein in hair regrowth and wound repair. The identified pOTBar-oleosin-rhFGF9 plasmid was transformed into Agrobacterium tumefaciens EHA105 by freeze-thaw method, and the oleosin-rhFGF9 gene was transformed into safflower leaves by A. tumefaciens mediated method. Transgenic safflower seedlings were then obtained by tissue culture. After basta screening, transgenic T₃ safflower seeds were obtained by grafting method, PCR verification and propagation. The expression of oleosin-rhFGF9 was detected by Western blot, and the content of oleosin-rhFGF9 fusion protein was 0.09% by using ELISA quantitative method. It was observed that 60 μg·L⁻¹ transgenic safflower oil had better effect on promoting NIH/3T3 cells proliferation in a certain dose-dependent manner. Sixty C57BL/6 mice were used to establish alopecia model and wound model respectively, and then were randomly divided into control group (treated with PBS or saline), negative group (treated with wild type safflower seed oil bodies, 60 g·L⁻¹), positive group (treated with FGF9, 0.054 g·L⁻¹), low dose group (treated with transgenic safflower oil bodies, 10 g·L⁻¹) and high dose group (treated with transgenic safflower oil bodies, 60 g·L⁻¹). The skin of all above-mentioned mice models were coated with soft adhesive manner every other day, 100 μL/time. After 15 days, the mice skin was cut and embedded for histological analysis. The hair regrowth experimental results showed that the hair of mice grew well, and the mice in high dose group had bushy hair, with significant effect on regeneration hair number as compared with the positive group. The healing was obvious in wound experiment, with significant healing effect in positive group, high dose group and low dose group as compared to blank control group. Furthermore, high dose group remarkably showed a better and higher healing effect than the positive group at day 5. Oleosin-rhFGF9 was successfully transformed into safflower, and T₃ transgenic safflower oil bodies expressed oleosin-rhFGF9 fusion protein were obtained, with the role of promoting hair regeneration and wound repair in mice.
ABSTRACT
BACKGROUND: A growth factor cocktail (GFC) including fibroblast growth factor 9 (FGF9) in combination with microneedling is an effective and safe treatment for patients with androgenetic alopecia (AGA). However, there is a lack of studies evaluating its effects based on microneedle depth. OBJECTIVE: This study aimed to evaluate the effects of a GFC including FGF9 on hair growth in patients with AGA, and compare the differences in efficacy according to microneedle depth. METHODS: The study was performed on patients with AGA who were treated with topical GFC including FGF9 with microneedling once every 2 weeks for 3 months. The scalp was divided into right and left sides, and treated with GFC including FGF9 (right side) and normal saline (left side). The microneedle depth was 0.8 mm for both sides. A total of 22 patients (11 males and 11 females) were enrolled. GFC including FGF9 was topically applied with a microneedle medical device. Treatment efficacy was evaluated through phototrichogram and digital photograph analyses after 6 repeated treatments for 3 months. RESULTS: The phototrichogram images showed that 3 months of treatment with GFC including FGF9 with microneedling increased hair density (27.4±4.4/cm²) and diameter (2.7±2.7 µm); increases in hair density (5.7±4.4/cm²) and diameter (2.2±2.3 µm) were also seen in the region of the scalp that received normal saline. These results were statistically significant (p < 0.05). The treatment effect was not significantly different between microneedle depths of 0.8 mm (used in this study) and 0.5 mm (used in our previous study) in terms of both hair density and hair diameter. CONCLUSION: GFC including FGF9 with microneedling is an effective and safe treatment for patients with AGA. According to the results of this study and our previous report, we believe that microneedle depths of 0.5∼0.8 mm can sufficiently stimulate the scalp to increase drug-delivery.