KIF18B通过激活Wnt_βcatenin信号通路促进子宫内膜癌细胞增殖和转移的实验研究.pdf

58现代检验医学杂志第 38 卷第 5 期2023 年 9 月J Mod Lab Med,Vol.38,No.5,Spet.2023KIF18B 通过激活 Wnt/catenin 信号通路促进 子宫内膜癌细胞增殖和转移的实验研究陆瑞，郭红霞，吴苗苗，吴志兵，姜雪，戈伍琼（南京中医药大学沭阳附属医院妇产科，江苏宿迁 223600）摘要：目的探讨驱动蛋白家族成员 18B（kinesin family member18b，KIF18B）在子宫内膜癌组织中的表达，及促进子宫内膜癌细胞增殖和转移的作用机制。方法收集子宫内膜癌组织 50 例和正常子宫内膜组织 30 例，采用免疫组织化学染色检测 KIF18B 表达水平，分析 KIF18B 表达与子宫内膜癌患者临床病理特征的关系及对患者预后的影响。KIF18B小干扰 RNA（siRNA）转染子宫内膜癌细胞 Ishikawa，分为 si-NC 组和 si-KIF18B 组，蛋白免疫印迹法检测 KIF18B siRNA 的转染效果；噻唑蓝比色法（MTT）检测细胞的增殖能力；Transwell 实验检测细胞的转移能力；双荧光素酶报告基因实验检测各组细胞中的 Wnt/-连环蛋白（-catenin）信号通路活性；蛋白免疫印迹法检测 Wnt/-catenin 信号通路相关蛋白 Wnt3a 和-catenin，及其下游增殖相关蛋白 cMYC，细胞周期蛋白 D1（cyclinD1）和转移相关蛋白基质金属蛋白酶 2（matrix metalloproteinase，MMP-2）、基质金属蛋白酶 7(matrix metalloproteinase-7，MMP-7)的表达。结果KIF18B在子宫内膜癌组织中的阳性表达率高于正常子宫内膜组织（70.00%vs 46.67%），差异有统计学意义（2=4.301，P=0.038）。FIGO 分期期、有淋巴结转移患者 KIF18B 阳性表达率高于 FIGO 分期期（88.89%vs 47.82%）、无淋巴结转移患者（86.36%vs 57.14%），差异有统计学意义（2=9.972,5.009；P=0.002，0.025）。KIF18B高表达组五年总生存率低于 KIF18B 低表达组（34.29%vs 66.67%），差异有统计学意义（2=4.305，P=0.038）。si-KIF18B 组 KIF18B 表达低于 si-NC 组（0.150.04 vs 1.010.03），差异有统计学意义（t=29.790，P0.001）。第 2 5天 si-KIF18B 组细胞增殖率低于 si-NC 组，差异均有统计学意义（t=3.267 11.080，均 P0.05）。si-KIF18B 组细胞穿膜数目低于 si-NC 组（39.674.52 个 vs 74.334.51 个），差异有统计学意义（t=9.402，P0.001）。si-KIF18B 组细胞中荧光素酶相对活性低于 si-NC 组（0.410.04 vs 1.000.03），差异有统计学意义（t=20.438，P0.001）。si-KIF18B组 Wnt3a，-catenin，cMYC，cyclinD1，MMP-2 和 MMP-7 表达低于 si-NC 组，差异均有统计学意义（t=4.69536.509，均 P 0.05）。结论KIF18B 在子宫内膜癌组织中表达增加，下调 KIF18B 可以抑制子宫内膜癌细胞的增殖和转移，KIF18B 可能通过激活 Wnt/-catenin 信号通路参与子宫内膜癌的发生、发展。关键词：子宫内膜癌；驱动蛋白家族成员 18B；Wnt/-连环蛋白；增殖；转移中图分类号：R737.33；R730.43文献标识码：A文章编号：1671-7414（2023）05-058-06doi:10.3969/j.issn.1671-7414.2023.05.011Experimental Study of KIF18B Promoting Endometrial Cancer Cell Proliferation and Metastasis by Activating Wnt/-catenin Signaling PathwayLU Rui，GUO Hongxia，WU Miaomiao，WU Zhibing，JIANG Xue，GE Wuqiong（Department of Obstetrics and Gynecology,Shuyang Affiliated Hospital of Nanjing University of Traditional Chinese Medicine,Jiangsu Suqian 223600,China）Abstract:ObjectiveTo investigate the expression of kinesin family member18b(KIF18B)in endometrial carcinoma and its mechanism of promoting proliferation and metastasis of endometrial carcinoma cells.Methods50 cases of endometrial cancer tissue and 30 cases of normal endometrial tissue were collected.Immunohistochemical staining was used to detect the expression level of KIF18B,and the relationship between KIF18B expression and clinical pathological parameters of endometrial cancer patients was analyzed,as well as its impact on patient prognosis.KIF18B small interfering RNA(siRNA)was transfected into endometrial cancer cell line Ishikawa and divided into si-NC group and si-KIF18B group.The transfection effect of KIF18B siRNA was detected by Western blotting.The proliferation ability of the cells was detected by thiazole blue colorimetry.Transwell assay was used to detect the ability of cell metastasis.The Wnt/-catenin signaling pathway activity in each group was detected by double luciferase reporter gene experiment.Wnt/-catenin signaling pathway related proteins Wnt3a and-catenin,as well as downstream proliferation-related proteins cMYC,cyclinD1 and transfer related proteins 基金项目：江苏省医学科研立项项目（编号：2020ZD028）：KIF18B 通过激活 Wnt/-catenin 信号通路促进了子宫内膜癌增殖和转移能力。作者简介：陆瑞（1968-），女，大学本科，主任医师，研究方向：妇科肿瘤，E-mail：。59现代检验医学杂志第 38 卷第 5 期2023 年 9 月J Mod Lab Med,Vol.38,No.5,Sept.2023matrix metalloproteinase(MMP-2)and MMP-7 were detected by Western blotting.ResultsThe positive expression rate of KIF18B in endometrial cancer tissue was higher than that in normal endometrial tissue(70.00%vs 46.67%),and the difference was statistically significant（2=4.301,P=0.038).The positive expression rate of KIF18B in patients with FIGO stage IIIIV and lymph node metastasis was higher than that in patients with FIGO stage III（88.89%vs 47.82%）and no lymph node metastasis(86.36%vs 57.14%),and the difference was statistically significant（2=9.972,5.009,P=0.002,0.025).The overall 5-year survival rate of the KIF18B high expression group was lower than that of the KIF18B low expression group(34.29%vs 66.67%),the difference was statistically significant(2=4.305,P=0.038).The expression of KIF18B in the si-KIF18B group was lower than that in the si-NC group(0.15 0.04 vs 1.01 0.03),and the difference was statistically significant(t=29.790,P0.001).On the 2nd to 5th day,the cell proliferation rate of the si-KIF18B group was lower than that of the si-NC group,and the differences were statistically significant(t=3.267 11.080,all P0.05).The number of cells penetrating the membrane in the si-KIF18B group was lower than that in the si-NC group(39.67 4.52 vs 74.33 4.51),the difference was statistically significant(t=9.402,P0.001).The relative activity of luciferase in the si-KIF18B group cells was lower than that in the si-NC group(0.41 0.04 vs 1.00 0.03),and the difference was statistically significant(t=20.438,P0.001).The expressions of Wnt3a,-catenin,cMYC,cyclinD1,MMP2 and MMP7 in si-KIF18B group were lower than those in si-NC grou