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Objective:To explore the effect of junctional plakoglobin (JUP) on the radiotherapy-resistant cervical cancer cells in vitro.Methods:Cervical cancer cell lines SiHa, HeLa and Me-180 were irradiated with 1 Gy of 60Co radioactive rays for 3 times every week to induce the radiotherapy resistance of cells which obtained the radiotherapy-resistant cell lines SiHaIR, HeLaIR and Me-180IR. The corresponding wild-type cell lines were served as control groups. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to detect the expressions of JUP mRNA and protein in the 6 groups of cells. The cell scratch test was used to detect the cell migration ability. Results:The relative expressions of JUP mRNA in SiHa and SiHaIR groups, HeLa and HeLaIR groups and Me-180 and Me-180IR groups were the relative expressions of JUP protein were 1.74±0.06 and 0.48±0.02 ( t = 12.327, P < 0.01), 1.77±0.06 and 0.28±0.03 ( t = 14.698, P < 0.01), 2.276±0.061 and 0.780±0.011 ( t = 7.367, P < 0.01); 2.36±0.03 and 0.55±0.02 ( t = 9.245, P < 0.01), 2.13±0.02 and 0.23±0.01 ( t = 15.643, P < 0.01), 1.96±0.05 and 0.73±0.02 ( t = 5.826, P < 0.01). When culturing the cells for 12, 24, and 48 h after scratching, the migration rates in 3 groups of radiotherapy-resistant cell lines were increased compared with the corresponding wild-type cell lines, and the differences were statistically significant (all P < 0.05). Conclusion:The expressions of JUP in radiotherapy-resistant cervical cancer cell lines are lower than those in wild-type cell lines, and the migration ability of radiotherapy-resistant cells is enhanced.
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Malignant ovariangerm cell tumors (MOGCTs) is second only to epithelial tumor which often occur in young women and young women,with high malignancy and high mortality.Effective treatment is particularly important in clinical practice.The prognosis is improved for valid chemotherapy scheme foun ded in recent years.Surgery still play a crucial role in the therapy of MOGCTs no matter for the primary operation or re-operation.Since the 70s,the comprehensive surgical staging (CSS) has improved the prognosis in patients with malignant ovarian cell tumors.Retroperitoneal lymphadenectomy is an integral part of the complete staging in MOGCT.The paper discusses various aspects of the clinical value of comprehensive surgical staging in MOGCTs.
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Objective To prepare a new-type acellular dermal matrix (ADM) and research on its relevant performance,which would provide theoretical evidence for clinical application.Methods Skin of Bama suckling pig was taken as resource of skin,and technologies of physics,chemistry and biology were selected to prepare new-type ADM.To detect the external structure,physical and chemical property as well as biological property of the prepared new-type ADM,hematoxylin-eosin (HE) staining observation,scanning electron microscope observation,amino acid analysis,material porosity and hydrophilicity test,tensile strength and in vitro degradation experiment,cytotoxicity test,and animal experiment have been conducted.Results New-type ADM cells have been thoroughly removed and dermal matrix remains intact with collagen content of 95.55％,connective three-dimensional pore structure,(85.03 ± O.99) ％ of porosity,(24.56 ± 0.57) ° of contact angle implying new-type ADM was hydrophilic substance,(5.48 ± 0.44) Pa of tensile strength implying its moderate level of pulling force,in vitro degradation period reduced to (28.7 ± O.76) h,and ＞75％ relative growth rate (RGR).Cells grew and proliferated on new-type ADM and could be replaced by original tissue after degradation.Conclusions New-type ADM have overcome disadvantages of traditional preparation method in sabotaging dermal matrix structure and incompletely removing cells from matrix,which is qualified with higher level of collagen content and porosity.With improved biological property,greatly reduced inflammation immunoreactions,and accelerated degradation rate,new-type ADM is of higher level of clinical application value.
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Objective To investigate the effects of specific methyltransferase inhibitor of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the promoter methylation of E-cadherin (E-cad) gene, protein expression in human cervical cancer SiHa cells, and the cell biological behavior. Methods SiHa cells were treated with 5-Aza-CdR at different concentrations. Quantitative methylation-specific polymerase chain reaction (QMSP) was used to examine CpG island promoter methylation level of E-cad gene before and after treatment. The experimental group of the optimum concentration was selected. The expression levels of E-cad mRNA and its protein in SiHa cells line were detected by quantitative real-time polymerase chain reaction (RT-PCR) and western blot respectively. Cell adhesion test was used to measure cell adhesion ability and Transwell test was used to detect cell invasion and migration ability. Results E-cad gene promoter methylation index (PMR) of 5-Aza-CdR at 0, 1, 5, 10, 15 μmol/L level was (53.0 ±1.6) %, (50.0 ±1.2) %, (44.0 ±1.4) %, (27.0 ±1.7) %, (15.0±8.2) %respectively, and PMR value decreased gradually with the increase of 5-Aza-CdR concentration. Furthermore, PMR value was the lowest at 15μmol/L, and the difference was statistically significant compared with other 4 groups (P< 0.01). Then 5-Aza-CdR at 15 μmol/L was selected as the following experimental concentration. The expression of E-cad mRNA and its protein in the 5-Aza-CdR group were significantly higher than those in the blank control group (P<0.05). The rates of cell adhesion , cell invasion inhibition and migration inhibition were all increased with significant differences (P<0.05). Conclusions 5-Aza-CdR can upregulate E-cad mRNA and protein expression level in cervical cancer SiHa cells, reduce cell invasion and migration ability, and promote the adhesion of SiHa cells, which has reversed hypermethylation in the promoter region of E-cad gene partly.
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Objective To investigate the effects of specific methyltransferase inhibitor of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the promoter methylation of E-cadherin (E-cad) gene, protein expression in human cervical cancer SiHa cells, and the cell biological behavior. Methods SiHa cells were treated with 5-Aza-CdR at different concentrations. Quantitative methylation-specific polymerase chain reaction (QMSP) was used to examine CpG island promoter methylation level of E-cad gene before and after treatment. The experimental group of the optimum concentration was selected. The expression levels of E-cad mRNA and its protein in SiHa cells line were detected by quantitative real-time polymerase chain reaction (RT-PCR) and western blot respectively. Cell adhesion test was used to measure cell adhesion ability and Transwell test was used to detect cell invasion and migration ability. Results E-cad gene promoter methylation index (PMR) of 5-Aza-CdR at 0, 1, 5, 10, 15 μmol/L level was (53.0 ±1.6) %, (50.0 ±1.2) %, (44.0 ±1.4) %, (27.0 ±1.7) %, (15.0±8.2) %respectively, and PMR value decreased gradually with the increase of 5-Aza-CdR concentration. Furthermore, PMR value was the lowest at 15μmol/L, and the difference was statistically significant compared with other 4 groups (P< 0.01). Then 5-Aza-CdR at 15 μmol/L was selected as the following experimental concentration. The expression of E-cad mRNA and its protein in the 5-Aza-CdR group were significantly higher than those in the blank control group (P<0.05). The rates of cell adhesion , cell invasion inhibition and migration inhibition were all increased with significant differences (P<0.05). Conclusions 5-Aza-CdR can upregulate E-cad mRNA and protein expression level in cervical cancer SiHa cells, reduce cell invasion and migration ability, and promote the adhesion of SiHa cells, which has reversed hypermethylation in the promoter region of E-cad gene partly.