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Objective To investigate the inhibitory effect of alantolactone on glioma C6 cells and its possible mechanism. Methods Rat glioma cell line C6 cells were cultured in vitro. The effect of AL on the viability of C6 cells was detected by MTT assay. The effect of AL on migration and invasion of C6 cells was assessed by scratch test and Transwell chamber assay. Flow cytometry was used to detect the effect of AL on the induction of apoptosis in C6 cells, and the effects of AL on mitochondrial membrane potential in C6 cells was detected using JC-1 fluorescent probe. Western blot was used to detect the expression of related proteins in C6 cells after AL treatment. Results AL significantly inhibited the proliferation of C6 cells in a time-and dose-dependent manner. After AL treatment for 24 hours, the migration and invasion of C6 cells were significantly inhibited, N-cadherin protein was significantly decreased and E-cadherin protein was significantly increased, the number of apoptotic cells increased obviously while the mitochondrial membrane potential decreased significantly. The protein expressions of PARP/ cleaved caspase-3/cleaved caspase-9 were significantly up-regulated. Conclusions AL inhibits the proliferation of glioma C6 cells by inhibiting their migration and invasion through regulating the protein expression of cadherin and inducing apoptosis in C6 cells by regulating the cytochrome C/caspase signaling pathway.
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Objective To observe exposure rates of hydroxyapatite artificial eye by comparing hydroxyapatite prosthesis implantation through lateral rhinotomy approach on the orbit and traditional sclera shell hydroxyapatite prosthesis implantation.Methods After eye content was enucleated at postoperative stage I,75 patients received hydroxyapatite prosthesis implantation,among which 26 received direct sclera shell prosthesis implantation (group A) and 49 received implantation through lateral rhinotomy approach on the orbit (group B).During postoperative follow-up from 6 months to 5 years,exposure rates of hydroxyapatite artificial eye under two operation ways were observed and analyzed by x 2 test.Results According to observation,8 cases were exposed in the group A,and 2 cases were repaired by fascia and conjunctival repair.Six cases were healed by prosthesis and sclera replacement.In group B,only 1 case was exposed slightly and recovered after simple conjunctival repair.The incidence of the eye exposure rate using two kinds of operation methods in group A is significantly higher than that in group B (P ＜ 0.05) The difference was statistically significant.Conclusion The exposure of hydroxyapatite artificial eye can be effectively avoided and postoperative complications can be reduced by hydroxyapatite prosthesis implantation through lateral rhinotomy approach on the orbit among patients with severe eyeball ruptures,painful eyeballs of no light perception and atrophy eyes.
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Objective To estimate the influence of 1,25(OH)2D3 on the proliferative ability of and methylation levels of genomic DNA and proliferation-associated gene promoter in human HaCaT keratinocytes.Methods Some cultured HaCaT cells were treated with 1,25 (OH)2D3 of 10-6,10-7 and 10-8 mol/L for 24 hours,then,methyl thiazolyl tetrazolium (MTT) assay was carried out to evaluate the proliferative activity of cells,and a global DNA methylation quantification kit was used to determine the global DNA methylation level.Real-time PCR was conducted to quantify the mRNA expression of DNA methyl transferases (DNMTs) and methyl-DNA binding domain (MBD) proteins,and methylation-specific PCR (MS-PCR) to evaluate the methylation status of promoter region in the programmed cell death 5 (PDCD5) and tissue inhibitor of metalloproteinase 2 (TIMP2) genes,in HaCaT cells after 24-hour treatment with 1,25 (OH)2D3 of 10-6 mol/L.The HaCaT cells receiving no treatment served as the control.Results Compared with the untreated HaCaT cells,those treated with 1,25(OH)2D3 of 10-6 mol/L showed significantly down-regulated proliferative activity (0.152 ± 0.027 vs.0.290 ± 0.017,P ＜ 0.01),global DNA methylation level (0.187 ± 0.071 vs.0.316 ± 0.049,P ＜ 0.05),DNMT3a and DNMT3b mRNA expression levels (P ＜ 0.01 or 0.05),but markedly upregulated mRNA expression levels of MECP2,MBD2,PDCD5 and TIMP2 (P ＜ 0.01 or 0.05).Moreover,the DNA methylation levels within the promoter region of PDCD5 and TIMP2 genes were significantly lower in HaCaT cells treated with 1,25 (OH)2D3 of 10-6 mol/L than in the control cells (0.38 ± 0.135 vs.0.72 ± 0.121,0.46 ± 0.172 vs.0.68 ± 0.133,both P＜ 0.05).Conclusions 1,25(OH)2D3 may down-regulate the global genomic DNA methylation level of,and modulate the expression of DNA methylationmodifying genes in,HaCaT cells.Furthermore,1,25 (OH)2D3 can decrease the promoter methylation levels but induce the overexpression of PDCD5 and TIMP2 genes,and decelerate the proliferation of HaCaT cells.