RESUMO
Objective:To investigate the role and mechanism of (histone deacetylase 6, HDAC6) in the epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells and the activation of renal interstitial fibroblasts.Methods:Human renal tubular epithelial cells (HK-2) and rat renal interstitial fibroblast (NRK-49F) were cultured in vitro, and divided into 4 groups: control group, Tubastatin A (TA) group (treated with 10 μmol/L HDAC6 inhibitor TA for 36 h), transforming growth factor-β1 (TGF-β1) group (10 ng/ml TGF-β1 for 36 h), and TGF-β1+TA group (treated with 10 ng/ml TGF-β1 and 10 μmol/L TA for 36 h). The expression levels of fibronectin, α-smooth muscle actin (α-SMA), collagen I, E-cadherin, HDAC6, acetyl histone H3, histone H3, acetyl α-tubulin, α-tubulin, TGF-β receptor (TGF-βR) 1, p-Smad3, Smad3, connective tissue growth factor (CTGF), epidermal growth factor receptor (EGFR) and p-EGFR in HK-2 and NRK-49F cell samples were detected by Western blotting, and quantitative analysis was performed according to gray level. Results:(1) In HK-2 cells stimulated by TGF-β1, TA decreased the expression of fibronectin, α-SMA, collagen I, and increased the expression of epithelial cell marker E-cadherin. Meanwhile, TA decreased the expression of HDAC6 and increased the expression levels of acetyl histone H3 and acetyl α-tubulin (all P<0.05). (2) Compared with the TGF-β1 group, the expressions of TGF-βR1, p-Smad3, CTGF and p-EGFR in TGF-β1+TA group were decreased (all P<0.05), while the total protein levels of Smad3 and EGFR were not significantly different (both P>0.05). (3) In NRK-49F cells stimulated by TGF-β1, TA decreased the expressions of fibronectin, α-SMA, collagen I, TGF-βR1 and p-Smad3 (all P<0.05). Conclusions:Blockade of HDAC6 by TA may inhibit the EMT of renal tubular epithelial cells and the activation of renal interstitial fibroblasts via regulating multiple signaling pathways including TGF-β/Smad3, CTGF and EGFR.
RESUMO
Objective:To study and investigate the expression effects of interleukin-8 (IL-8)in interstitial fibroblast cells of breast cancer before and after the application of docetaxel chemotherapy.Methods:46 cases of breast cancer without chemotherapy surgical resection in patients with resected tissue specimens from August 2014 to February 2016 in our hospital were selected,all specimens were cultured to form stable breast cancer stromal fibroblast cells,each cell line after raising Daipei were divided into the observation group and control group,the control group cells without any treatment,the observation group of 24h cells treated with docetaxel(20ng/mL),two groups of cells were extracted from the mRNA and analyzed the gene expression and differential gene expression analysis,and then to PCR and protein to detected the expression level of electrophoresis detection of interleukin two interleukin-8 cells,the expression of interleukin-8 in two groups of breast cancer cells was observed and compared.Results:After docetaxel chemotherapy,the breast cancer stromal fibroblast gene expression changed,and compared to the control group,the observation group were given docetaxel chemotherapy after breast cancer between matter into fiber cell interleukin-6 serum IL-8 mRNA and protein expression levels were significantly higher than that of the control group,the difference had statistical significance (P＜0.05).Conclusions:Interleukin IL-8 in the breast cancer matter into fiber cells were significantly up-regulated after docetaxel chemotherapy,indicating that the expression changes of breast cancer may be related with the effect of chemotherapy and chemotherapy drug resistance related.