SIX4在子宫内膜癌中的表达与临床意义及其对Ishikawa细胞侵袭和迁移的影响

Expression and Clinical Significance of SIX4 in Endometrial Carcinoma and its Effect on Invasion and Migration of Ishikawa Cells

目的探究同源异型盒基因4(sine oculis homeobox homolog 4,SIX4)在子宫内膜癌组织中的表达、临床意义及其对Ishikawa细胞侵袭和迁移的影响。方法采用实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)法检测SIX4在子宫内膜癌组织(内膜癌组)及正常内膜组织(对照组)的表达水平,并分析其与内膜癌组患者临床病理特征的相关性;将Ishikawa细胞分为正常组(空白对照组)、阴性对照组(转染siRNA-NC组)和抑制组1(转染siRNA-1组),抑制组2(转染siRNA-2组),抑制组3(转染siRNA-3组),RT-qPCR法检测各组SIX4表达量。Western blot法检测各组细胞SIX4、E-cadherin及N-cadherin的蛋白表达水平;Transwell实验、划痕实验分别检测各组细胞侵袭和迁移能力。结果与对照组比较,SIX4在内膜癌组中的表达水平较高(P<0.05),且SIX4高表达与子宫内膜癌的临床分期、肌层浸润深度、淋巴结转移情况相关(P<0.05),但与年龄、组织类型和分化程度均无关(P>0.05);siRNA抑制SIX4表达后,RT-qPCR法检测结果显示,抑制组2干扰效果最佳。Western blot法检测结果显示,抑制组相较于正常组及阴性对照组E-cadherin表达升高,SIX4和N-cadherin表达降低。Transwell实验及划痕实验结果显示,抑制组细胞的侵袭和迁移能力均较正常组及阴性对照组显著降低(P<0.05)。结论SIX4在子宫内膜癌组织中呈现高表达,其高表达与子宫内膜癌的不良病理特征相关;siRNA靶向抑制SIX4的表达可通过上皮间充质转化通路抑制子宫内膜癌细胞的侵袭和迁移。

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(transfected 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;Transwell 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 inhibition 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 experiments 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.