摘要
目的研究己糖激酶2(HK2)对宫颈癌细胞HeLa增殖和迁移的影响及其作用机制。方法G418压力筛选获取HK2稳定表达的HeLa细胞系;细胞计数法、MTT法、流式细胞仪检测HK2对HeLa细胞增殖、细胞活力及细胞周期的影响;细胞划痕和Transwell小室检测HK2对HeLa细胞迁移和侵袭能力的影响;Western blot和免疫细胞化学检测β-catenin、Cyclin D1、MMP7和Slug蛋白的表达;TOP/FOP实验检测HK2对Wnt/β-catenin信号转导通路活性的影响;Wnt/β-catenin信号通路抑制剂XAV-939观察在HK2过表达HeLa细胞中抑制Wnt/β-catenin信号通路活性后细胞增殖、迁移和侵袭能力的变化。结果成功构建HK2稳定表达的HeLa细胞系;过表达HK2可以促进HeLa细胞的增殖和细胞活力、促进细胞周期从G_0/G_1期向S期转换、增强细胞迁移和侵袭能力、增强Wnt/β-catenin信号转导通路活性,上调Cyclin D1、MMP7和Slug的表达;XAV-939抑制Wnt/β-catenin信号通路在HK2过表达HeLa细胞中的活性后,可以抑制HK2对细胞增殖、迁移和侵袭的促进作用。结论HK2通过Wnt/β-catenin信号转导通路上调Cyclin D1、MMP7和Slug蛋白表达促进HeLa细胞增殖、迁移和侵袭。
Objective To investigate the effect of HK2 on the proliferation and migration of cervical cancer cell lines HeLa and related mechanism.Methods HeLa cells with stable HK2 expression were screened.Cell proliferation,viability and cell cycle were detected by cell growth curves,MTT assay and flow cytometry,respectively.Wound-healing assay and Transwell cell invasion assay were used to detect the effect of HK2 on the migration and invasion potential of HeLa cells.The expression ofβ-catenin,Cyclin D1,MMP7 and Slug were detected by Western blot and immunocytochemistry.The activity of Wnt/β-catenin signalling pathway was detected by TOP/FOP assay.Cell growth,migration and invasion were detected in HeLa-HK2 cells treated with Wnt/β-catenin inhibitor(XAV-939).Results HeLa cells lines that stably expressing HK2 were successfully constructed.HK2 overexpression promoted the proliferation,migration,invasion and viability of HeLa cells,accelerated the transition of cell cycle from G0/G1 phase to S phase,enhanced the activity of Wnt/β-catenin signalling pathway and up-regulated the protein expression of downstream factors,such as Cyclin D1,MMP7 and Slug.The promoted growth,migration and invasion of HeLa-HK2 cells were inhibited by Wnt/β-catenin inhibitor XAV-939.Conclusion HK2 could promote the proliferation,migration and invasion of HeLa cells by up-regulating the expression of Cyclin D1,MMP7 and Slug via Wnt/β-catenin signaling pathway.
作者
冯倩
刘宪
张妮
崔南
FENG Qian;LIU Xian;ZHANG Ni;CUI Nan(Department of Reproductive Medicine,The First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China;Medical College of Xi’an Jiaotong University,Xi’an 710061,China;Department of Cell and Genetics,Basic Medicine,Xi’an Medical University,Xi’an 710021,China)
出处
《肿瘤防治研究》
CAS
CSCD
2020年第9期649-654,共6页
Cancer Research on Prevention and Treatment
基金
国家自然科学基金(81903042)。
