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Objective:To investigate the effect of different chemotherapy drugs combined with DNA methylase inhibitor 5-Aza-2'-deoxycytidine (5-Aza-dC) on the apoptosis of lung adenocarcinoma cells.Methods:In the prospective randomized controlled study, lung adenocarcinoma A549 cells were treated with cisplatin plus paclitaxel (TP) or gemcitabine (GP) with or without 5-Aza-dC. According to different drug intervention methods, they were divided into control group, cisplatin combined with paclitaxel (TP) group, cisplatin combined with gemcitabine (GP) group, and 5-Aza-dC combined with TP group, 5-Aza-dC combined with GP group. CCK-8 assay was used to detect the proliferation of A549 cells. Transwell migration and invasion assay were used to detect the effect that each group of drugs on the migration and invasion ability of A549 cells. Quantitative Real-time Polymerase Chain Reaction was used to evaluate the effect of each treatment on the expression of apoptotic genes. One-way analysis of variance was used to compare the degree of cell proliferation in different drug treatment groups, and LSD- t method was used for pairwise comparison within groups. Results:The inhibition rates of lung adenocarcinoma cells in the TP regimen at different time points at 24, 48, and 72 h were as follows (20.00±4.23) %, (35.00±2.80) %, and (56.00±3.11) %. The inhibition rate of 5-Aza-dC combined with TP regimen on lung adenocarcinoma cells was significantly increased, at different time points of 24, 48 and 72 h, respectively (38.00±3.80) %, (50.00±3.25) %, (93.00±4.33) %. The inhibition rates of cells at different time points at 24, 48, and 72 h in the GP regimen were (33.00±5.10) %, (54.00±3.80) %, and (74.00±2.82) %, respectively; while 5-Aza-dC combined with GP regimen could significantly reduce the rate of cell growth, the inhibition rates of cells at 24, 48, and 72 h different time points were as follows (54.00±3.00) %, (67.00±5.30) %, and (95.00±1.13) %. The inhibitory effect of the same drug on lung adenocarcinoma cells increased with time (TP group: F=35.93, P<0.001; 5-Aza-dC combined with TP group: F=97.33, P<0.001; GP group: F =41.73, P<0.001; 5-Aza-dC combined with GP group: F=79.00, P<0.001), and at different time points, the differences were statistically different (all P<0.05). 5-Aza-dC combined with TP and GP chemotherapy regimens can inhibit the migration and invasion of lung adenocarcinoma cell A549, and the inhibitory effect is stronger than that of TP or GP regimens alone. The expression of Caspase 8 was significantly elevated ( t=5.87, P=0.004) in cells treated with 5-Aza-dC combined with GP when compared with GP regimen alone. The expression of Caspase 8 ( t=3.94, P=0.017), Caspase 6 ( t=5.81, P=0.004) and BBC3 (BCL-2 binding component 3) ( t=6.53, P=0.003) were increased when drugged with 5-Aza-dC combined TP regimen compared with TP regimen alone. Conclusion:5-Aza-dC might serve as a chemotherapeutic sensitizer to increase the sensitivity of lung adenocarcinoma cells.
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Objective To observe the acute side effects and recent therapeutical effect of cisplatinum/paclitaxe concurrent radiotherapy for patients with advanced nasopharyngeal carcinoma.Methods 87 patients with advanced nasopharygeal carcinoma were enrolled.According to the treatment,87 patients were divided into paclitaxe group (groupA,n =43) and cisplatinum group (group B,n =44).The two groups were treated with 6MV-X ray for radiotherapy.The prescription dose of radiation therapy was PGTV,PGTVnd 6 996 cGy/33F,PTV1 6 006 cGy/33F,PTV2 5 096 cGy/28F.The two groups began concurrent chemotherapy in radiotherapy,specific method:group A paclitaxe 30mg/m2,1 time a week,until the end of radiotherapy,group B cisplatinum 80 ～100 mg/m2,every 21 days.Results The recent therapeutical effect of nasopharyngeal lesions and lymph nodes of neck for group A and group B were 88.4％ and 86.4％ (x2 =0.079,P =0.778),88.4％ and 88.6％ (x2 =0.000,P =1.000),respectively.But the occurrence rate of group B ' oral mucosa reaction (x2 =4.295,P =0.038) and gastrointestinal reaction (x2 =4.482,P =0.028) were higher than that of group A.Conclusion Treated with cisplatin in advanced nasopharyngeal carcinoma after radical radiotherapy combined with docetaxel is similar,and the side effect is low,the tolerance of patients is good,curative effect is satisfied.
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Background The excessive growth of human Tenon fibroblasts (HTFs) is a primary cause of failure of anti-glaucomatous filtering surgery.To seek a drug of anti-fibrosis is of an important significance for improving the successful rate of anti-glaucomatous filtration surgery.Objective The goal of this study was to investigate the effect of paclitaxel on proliferation of HTFs in vitro.Methods Human Tenon tissue was obtained during the anti-glaucomatous filtering surgery.HTFs were cultivated using explant method and 3-6 generations of cells were used in the experiment.The cells were identified by immunochemistry using keratin,vimentin,fibronectin and S-100.Different concentrations (0,1 ×10-s,1 × 10-7,1 × 10-6 mol/L) of paclitaxel were added into the medium,and then the cell indexes (CI) in the various groups were detected by real-time cell electronic sensing (RT-CES) 24 hours after affection of paclitaxel.Apoptosis of the cells was examined using DAPI staining,and the proportion of the cells in different cycles were assayed by flow cytorneter 12 hours after addition of paclitaxel.The expressions of matrix metalloproteinase-1 (MMP-1) protein and mRNA were detected by ELISA and real-time PCR,respectively.Results The cells migrated from the cultivated tissue within 7 days with the fibrocyte-like shape.The cells showed the positive response for vimentin and absent response for keratin,fibronectin and S-100.The CI values were 1.093 ±0.191,0.665 ± 0.093 and 0.473 ± 0.117 in the 1 × 10-8,1 × 10-7 and 1 × 10-6 mol/L paclitaxel groups,showing significant rise in comparison with the 1.514 ±0.283 of the 0 mol/L paclitaxel group (all at P =0.000).The cell nuclei were normal in the 0 mol/L paclitaxel group,however,blue-fluorescent particles and apoptotic bodied were found in the cell nuclei after affection of paclitaxel.The proportion of G2/M phase of cells were (9.20±0.80) ％,(12.37±0.45)％ and (13.80±0.35)％ in the 1×10-8 mol/L,1×10-7 mol/L and 1×10-6 mol/L paclitaxel groups,which were higher than the (7.17±0.50) ％ in the 0 mol/L paclitaxel group (P=0.005,0.000,0.000).In addition,the relative expressing level of M MP-1 mRNA (MMP-1 mRNA/GAPDH mRNA) and the expression level of MMP-1 protein in the HTFs were significantly lower in the 1 ×10-8 mol/L,1 × 10-7 mol/L and 1 × 10-6 mol/L paclitaxel groups than those in the 0 mol/L group (all at P＜0.05).Conclusions Paclitaxel at the concentrations of 1 × 10-8 mol/L-1 × 10-6 mol/L inhibits the proliferation of HTFs in vitro by arresting the cellular mitosis and inducing cell apoptosis.These effects probably associated with down-regulation of MMP-1 expression in the HTFs.
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Objective:To investigate the effect and potential mechanism of combined paclitaxe and tumor necrosis factor-related poptosis-inducing ligand(TRAIL) on apoptosis of human gastric cancer cell in vitro.Methods:SGC7901 cells were cultured with paclitaxe at different concentrations(1,5,10,20,40?g/ml) and different time(12,24,36,48h).Methyl thiazolyl tetrazolium (MTT) assay was used to measure the anticancer activity of paclitaxe.The ability of TRAIL alone,paclitaxe alone and TRAIL in combination with paclitaxe to induce apoptosis was measured by a flow cytometer.Expression of DR4,DR5 mRNA was examined by RT-PCR.Results:The apoptosis rate of control group(C group),paclitaxe group(P group),TRAIL group(T group) and combination group(T+P)in 24h were 2.09%,10.65%,7.79% and 24.51% respectively.T+P group was significantly higher than that in C group,T group and P group(P
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Aim To investigate the effect of exposure to high or low concentration chemotherapeutic drugs on multidrug resistance of human breast cancer cell MDA-MB-231.Method MDA-MB-231 was treated with high dose of adriamycin and paclitaxel or with low concentration of paclitaxe.SRB assay was used to determine the IC50 and RF.HE assay was used to evaluate the cellular morphology.The variations of P-gp and MDR1 were analyzed by immunocytochemistry and RT-PCR respectively.Results Cells survived only after treated with high dose of paclitaxel (MDA-MB-231/a).SRB assay showed that the growth rate of MDA-MB-231/a was the same as that of parent MDA-MB-231/w.The IC50 of the cells(MDA-MB-231/b)treated with low concentration of paclitaxel to several chemotherapeutic drugs was a little higher than that of MDA-MB-231/w.Immunocytochemistry showed that there was no difference between MDA-MB-231/a and MDA-MB-231/w in the expression of P-gp while the P-gp expression was a little higher in MDA-MB-231/b.RT-PCR assay showed that the MDR1 gene was over-expressed in MDA-MB-231/b.Conclusions The multidrug resistance cell lines can not be derived from MDA-MB-231/w by high dose of chemotherapeutic drugs.Induction of multidrug resistance response to chemotherapeutic drugs is related with transient exposure to low concentration of paclitaxel and this may be a way to establish the multidrug resistance model of MDA-MB-231 cells.