CAJ | 학술논문

目的：探讨DKK1 siRNA干扰的DKK1低表达对子宫内膜癌Ishikawa细胞侵袭、迁移能力的影响及作用机制。方法瞬时转染DKK1 siRNA至培养的子宫内膜癌Ishikawa细胞；应用实时定量PCR和蛋白免疫印迹方法分别检测转染前后细胞中DKK1的mRNA和蛋白表达；应用Transwell实验检测癌细胞侵袭、迁移能力。应用荧光显微镜观察转染前后Wnt/β-catenin信号通路中活性β-catenin、MMP14表达情况。结果转染DKK1 siRNA至Ishikawa细胞使细胞中DKK1基因表达水平降低21.00%（t＝3.05，P＜0.05）， DKK1蛋白表达降低39.35%（t＝40.97，P＜0.01）。侵袭实验中DKK1 RNAi组的细胞均数140.8±4.733，高于空白对照组的细胞均数123.7±6.700，癌细胞的侵袭能力增强13.82%。迁移实验中DKK1 RNAi组细胞均数（152.0±3.528）高于空白对照组（130.6±4.061），癌细胞的迁移能力增强16.39%。DKK1 siRNA处理后，细胞中活性β-catenin、MMP14荧光增强，提示表达水平上升。结论 DKK1具有抑制子宫内膜癌Ishikawa细胞侵袭、迁移的作用。Wnt/β-catenin信号通路参与了DKK1这一作用过程。DKK1不失为抑制子宫内膜癌转移的有效治疗靶点。
목적：탐토DKK1 siRNA간우적DKK1저표체대자궁내막암Ishikawa세포침습、천이능력적영향급작용궤제。방법순시전염DKK1 siRNA지배양적자궁내막암Ishikawa세포；응용실시정량PCR화단백면역인적방법분별검측전염전후세포중DKK1적mRNA화단백표체；응용Transwell실험검측암세포침습、천이능력。응용형광현미경관찰전염전후Wnt/β-catenin신호통로중활성β-catenin、MMP14표체정황。결과전염DKK1 siRNA지Ishikawa세포사세포중DKK1기인표체수평강저21.00%（t＝3.05，P＜0.05）， DKK1단백표체강저39.35%（t＝40.97，P＜0.01）。침습실험중DKK1 RNAi조적세포균수140.8±4.733，고우공백대조조적세포균수123.7±6.700，암세포적침습능력증강13.82%。천이실험중DKK1 RNAi조세포균수（152.0±3.528）고우공백대조조（130.6±4.061），암세포적천이능력증강16.39%。DKK1 siRNA처리후，세포중활성β-catenin、MMP14형광증강，제시표체수평상승。결론 DKK1구유억제자궁내막암Ishikawa세포침습、천이적작용。Wnt/β-catenin신호통로삼여료DKK1저일작용과정。DKK1불실위억제자궁내막암전이적유효치료파점。
Objective The purpose of this study was to investigate the influence mechanism of siRNA-mediated DKK1 low expression on invasion and migration abilities of endometrial carcinoma Ishikawa cell lines. Methods Cultured endometrial carcinoma Ishikawa cell lines were transient transfected by DKK1 siRNA;mRNA and protein levels of DKK1 in the transfected cells and untransfected cells were detected by real time PCR and western blot analyses, respectively. Invasion and migration abilities of the transfected cells were detected by Transwell assay. Expression location and profiles of activeβ-catenin and MMP14 of Wnt/β-catenin signal pathway in cells were detected by immunofluorescence. Results Expression of DKK1 gene in cells transfected with DKK1 siRNA was down-regulated by 21.00 percent (t=3.05, P<0.05). Expression of DKK1 protein in cells transfected with DKK1 siRNA was down-regulated by 39.35 percent (t=40.97, P<0.01). In invasion assay, average cell number in DKK1 siRNA group (140.8±4.733) was more than that in Blank group (123.7±6.700). The invasion ability of the cells increased by 13.82 percent as compared to Blank group. In migration assay, average cell number in DKK1 siRNA group (152.0±3.528) was more than that in Blank group (130.6±4.061). The migration ability of the cells increased by 16.39 percent as compared to Blank group. After DKK1 siRNA transfection, the expression of activeβ-catenin and MMP14 were up-regulated immunofluorescence expression. Conclusion DKK1 can inhibit the invasion and migration abilities of endometrial carcinoma Ishikawa cell lines. Wnt/β-catenin signal pathway involved in the process. DKK1 may be a good target for anti-metastasis treatment in endometrial carcinoma.