1.Effects of mitochondrial fission in proliferation, apoptosis and invasion of thyroid squamous carcinoma cell line SW579
Feifei SHEN ; Rui GUO ; Shupeng ZHAO ; Fengjie QI ; Cuili MENG
Tianjin Medical Journal 2015;(2):129-132,133
Objective To detect the expression of mitochondrial dynamics proteins (Mfn2 and Drp1) in thyroid squa?mous carcinoma cell line SW579 and the effects of Mitochondrial division inhibitor, Mdivi-1, on proliferation, apoptosis and invasion of SW579. Methods In SW579 and Nthy-ori 3-1 cell lines, the expression levels of Mfn2 and Drp1 were deter?mined by western blot while the transcription level of Mfn2 and Drp1 mRNA were measured by RT-PCR. Then, SW579 cells were divided into control group (DMSO, 0.1%) and Mdivi-1 low, medium and high dose groups (Mdivi-1 of 15,30 and 45μmol/L were incubated with cells for 16 hours respectively). Then the ability of cell proliferation was detected using MTT assay, the mitochondrial membrane potential was determined by fluorescence spectrophotometer, the expression levels of cy?tochrome C and Caspase-3 were quantified by Western blot and the transcription level of the Cyt C and Caspase-3 mRNA were determined by RT-PCR. The ability of invasion in each group was measured with Transwell assays. Results Com?pared with Nthy-ori 3-1, the mRNA transcription and protein expression levels of the Mfn2 was remarkably decreased, while the mRNA transcription and protein expression of the Drp1 was significantly increased in SW579 cells (P<0.01). Compared with control group, the cell survival rates and mitochondrial membrane potential of SW579 were decreased dramat?ically (P<0.01). The mRNA transcription and protein expression of the cytochrome C and Caspase-3 were increased dra?matically (P<0.01) and the capability of invasion was markedly decreased in all the Mdivi-1 groups in a dosage dependent manner compared with those in control groups (P<0.01). Conclusion Abnormal mitochondrial dynamics may be involved in thyroid squamous cell carcinoma SW579 cells;Mdivi-1 can inhibit the cell proliferation and invasion as well as induce apoptosis.
2.Graphene oxide promotes diabetic skin wound healing through regulating adipose-derived mesenchymal stem cells
Fengjie SHEN ; Weinan CAO ; Zhe JI ; Xinhua ZHANG ; Dongmei ZHOU
Chinese Journal of Endocrinology and Metabolism 2023;39(9):790-796
Objective:To study the effect of graphene oxide (GO) on adipose-derived mesenchymal stem cells (Ad-MSCs) to promote diabetic skin wound healing.Methods:Ad-MSCs were cultured with GO at concentration of 100, 200, and 400 μg/mL and with 25 mmol/L of glucose for 48 h. The apoptosis of Ad-MSCs was detected by flow cytometry and Western blotting. Eighteen BALB/c thymus free female nude mice were selected to establish the skin defect model of diabetic nude mice. Then phosphate-buffered saline, Ad-MSCs, and GO+ Ad-MSCs were injected into the damaged skin of nude mice by intradermal injection for treatment, thereby created the control group, Ad-MSCs group, and GO+ Ad-MSCs group, 6 mice each group. The survival of wound cells and skin healing of mice were observed after surgery, and the wound healing was determined by HE staining and Masson staining.The level of epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), tumor necrosis factor-α (TNF-α), and interleukin 6 (IL-6) were determined by enzyme-linked immunosorbent assay (ELISA).Results:Compared with the high glucose group without GO(control group), the apoptosis rate of Ad-MSCs co-culture with GO was significantly reduced( P<0.05), and the apoptosis rate decreased with the increase of GO concentration. The expression level of Bax protein decreased, and the expression level of Bcl-2 protein increased ( P<0.05). Diabetic skin defect model showed that the survival rate of Ad-MSCs and wound healing degree in GO+ Ad-MSCs group were significantly increased ( P<0.05), and the degree of epithelialization and thickness of collagen regeneration were better than those in other groups. ELISA results showed that the expressions of EGF and VEGF were significantly increased in GO+ Ad-MSCs group ( P<0.05), while the expressions of TNF-α and IL-6 were significantly decreased ( P<0.05). Conclusion:GO can inhibit the apoptosis of Ad-MSCs in vitro. In vivo, Ad-MSCs treated with GO have higher survival rate, faster wound healing, and better effect than Ad-MSCs treated alone.