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Objective To explore the expression and clinical significance of sine oculis homeobox homolog 4(SIX4)in endometrial carcinoma and its effect on the invasion and migration of Ishikawa cells.Methods Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of SIX4 in endometrial carcinoma tissue(endometrial carcinoma group)and normal endometrial tissue(control group).The correlation between the expression level and the clinicopathological characteristics of endometrial carcinoma patients was analyzed.Ishikawa cells were divided into normal group(blank control group),negative control group(transfect-ed with siRNA-NC group)and inhibition group 1(transfected with siRNA-1 group),inhibition group 2(transfected with siRNA-2 group),inhibition group 3(transfected with siRNA-3 group),RT-qPCR was used to detect the expression of SIX4 in each group.Western blot was used to detect the protein expression level of SIX4,E-cadherin and N-cadherin in Ishikawa cells of each group;Tran-swell test and scratch test were used to detect the invasion and migration ability of Ishikawa cells in each group.Results Compared with the control group,the expression level of SIX4 was higher in endometrial carcinoma group(P<0.05).The high expression of SIX4 was correlated with the clinical stage of endometrial carcinoma,the depth of muscular invasion and lymph node metastasis(P<0.05),but not with age,tissue type and differentiation degree(P>0.05).After SIX4 expression was inhibited by siRNA,RT-qPCR showed that inhi-bition group 2 had the best interference effect.Western blot showed that the expression of E-cadherin in inhibition group was higher than that in normal group and negative control group,while the expression of SIX4 and N-cadherin was lower.Transwell and scratch experi-ments showed that the invasion and migration of cells in the inhibition group were significantly lower than those in the normal group and negative control group(P<0.05).Conclusion SIX4 is highly expressed in endometrial carcinoma tissues,and its high expression is related to the adverse pathological features of endometrial carcinoma.siRNA targeted inhibition of SIX4 expression can inhibit the invasion and migration ability of endometrial cancer cells through epithelial mesenchymal transition pathway.
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Objective To study the effects of anti-oncogene WWOX on cell growth of epithelial ovarian cancer,in order to find a new approach of gene therapy for ovarian cancer.Methods A eukaryotic expression vector containing WWOX was transfected into ovarian cancer cell line HO8910 in vitro (recombinant plasmid group),and positive cell clones were selected and amplified.Expression of WWOX protein was detected by western blot. Untransfected cell(blank contrast group) and transfected empty plasmid cell(empty plasmid group)were served as control groups.In vitro,the biology effect of WWOX on HO8910 cell was analyzed throush the methyl thiazolyl tetrazolium test,transwell chamber cell invasion assay in vitro,agarose clony-formation and flow cytometry.In vivo,the cell of transfection was transplanted intraperitoneally in to BALB/c nude mice.The survival time and growth ability of nude mice were observed.Results (1)Recombinant plasmid group cell could steadily express WWOX protein,while in empty plasmid group and blank control group the expression of WWOX protein were not detected.(2)The growth rate of recombinant plasmid group cell was inhibited.(3)The agnrose clony-formation rate of recombinant plasmid group(19.8%)was significantly lower than that of the empty plasmid group(54.5%)and blank control group(56.0%,P＜0.05).(4)Flow cytometry showed that(72.08±0.39)% of cells was arrested at G0/G1 stage in recombinant plasmid group, while in empty plasmid group and blank control group G0/G1 stage cells were at (41.02±1.08)% and (39.31±0.67)% (P＜0.05). (5) In vitro invasion assay showed that invasion cell number in recombinant plasmid group (89.7±3. 1 ) was not significantly different from that of empty plasmid group(91.2±1.3) and blank control group(91.4±1.3, P ＞0. 05). (6) In vivo test in nude mice showed that WWOX gene could inhibit tumor growth of the HO8910 cells. Conclusions Tumor suppressor gene WWOX could interfere with the cell cycles of ovarian cancer cell and inhibit cell proliferation. As a new valuable tool,it premises to have application in the gene therapy of ovarian cancer.
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Objective To assess the clinical significance of changes tumor necrosis factor(TNF)-alpha level and the relationship between serum TNF-? and glucose levels in patients with gestational diabetes mellitus(GDM). Methods 71 patients with gestational diabetes mellitus were enrolled.40 normal pregnant women were assigned to be control group.Then the concentrations of blood glucose and TNF-? were determined. Results Blood TNF-? level in GDM patients was signficantly higher than that in normal pregnant women(P