HAX-1调控人宫颈癌Hela的细胞增殖  

HAX-1 regulates the proliferation of human cervical carcinoma Hela cells

作　　者：季瑞[1] 唐敏 徐海波[1] 姚涓 陆云燕[1] 金华[2,3] JI Rui;TANG Min;XU Haibo;YAO Juan;LU Yunyan;JIN Hua(Department of Gynecological Oncology,Tumor Hospital Affiliated to Nantong University,Nantong 226361,Jiangsu;Central Laboratory,Tumor Hospital Affiliated to Nantong University,Nantong 226361,Jiangsu;Nantong University Medical School,Nantong 226001,Jiangsu,China)

机构地区：[1]南通大学附属肿瘤医院妇瘤科,江苏南通226361 [2]南通大学附属肿瘤医院中心实验室,江苏南通226361 [3]南通大学医学院,江苏南通226001

出　　处：《临床检验杂志》2023年第5期374-379,共6页Chinese Journal of Clinical Laboratory Science

基　　金：江苏省高层次卫生人才“六个一工程”拔尖人才科研项目(LGY2020050);南通市科技计划指导性项目(JCZ20016);南通市卫建委项目面上课题(MB2020023)。

摘　　要：目的探讨造血细胞特异性蛋白1(HS-1)相关蛋白X-1(HAX-1)对宫颈癌Hela细胞增殖的调控作用。方法以人宫颈癌Hela细胞系为研究模型,采用脂质体介导法将HAX-1 siRNAs和阴性对照siRNA转染Hela细胞并构建HAX-1沉默细胞系(HAX-1沉默组),以HAX-1高表达质粒和pcDNA3.1空载质粒转染Hela细胞并构建HAX-1过表达细胞系(HAX-1过表达组)。采用实时荧光定量PCR(RT-qPCR)和Western blot检测转染后Hela细胞中HAX-1的表达情况;EdU试验检测转染后的细胞增殖能力;MTT试验检测转染后的细胞活力;流式细胞术检测HAX-1对转染后Hela细胞周期和细胞凋亡的变化;RT-qPCR检测转染后Ki-67、c-Myc、BCL-2/BAX基因的表达水平。结果沉默HAX-1后,Hela细胞中HAX-1 mRNA和蛋白质的水平显著降低,而过表达HAX-1后,Hela细胞中HAX-1 mRNA和蛋白质的水平显著升高,差异均有统计学意义(P<0.05)。过表达HAX-1后,HAX-1过表达组中EdU阳性细胞数目下降,细胞活力降低,G1期细胞比例增加,S期细胞比例降低,细胞凋亡率上升,增殖相关基因Ki67表达降低,BCL-2/BAX基因显著降低(P<0.05),而HAX-1沉默组则具有相反的表现。结论HAX-1的表达抑制了Hela细胞增殖,并通过BCL-2/BAX途径诱导其凋亡。Objective To investigate the regulatory effect of hematopoietic cell-specific protein 1-associated protein X-1(HAX-1)on the proliferation of cervical cancer Hela cells.Methods Using a human cervical cancer Hela cell line as the research model,HAX-1 siRNAs and negative control siRNA were transfected into Hela cells by the liposome mediated method to construct a HAX-1 silencing cell line(HAX-1 silencing group).The HAX-1 over-expression plasmid pcDNA3.1 was transfected into Hela cells to construct a HAX-1 over-expression cell line(HAX-1 over-expression group).The expression levels of HAX-1 in transfected Hela cells were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blot.The cell proliferation ability and viability after transfection were detected by the EdU assay and MTT assay,respectively.The changes of Hela cell cycle and apoptosis after transfection were determined by flow cytometry.The expression levels of Ki-67,c-Myc and BCL-2/BAX genes after transfection were detected by RT-qPCR.Results After silencing HAX-1,the levels of HAX-1 mRNA and protein in Hela cells were significantly reduced,while the over-expression of HAX-1 significantly increased the levels of HAX-1 mRNA and protein in Hela cells,with statistical significance(P<0.05).The over-expression of HAX-1 led to decreased number of EdU positive cells,decreased cell viability,increased proportion of cells in G 1 phase,decreased proportion of cells in S phase,increased apoptosis,and decreased expression levels of Ki67 and BCL-2/BAX genes(P<0.05),while the HAX-1 silencing group showed the opposite performance.Conclusion The expression of HAX-1 may inhibit the proliferation of Hela cells and induce their apoptosis by the BCL-2/BAX pathway.

关 键 词：造血细胞特异性蛋白1相关蛋白X-1 宫颈腺癌 凋亡 细胞增殖 

分 类 号：R446[医药卫生—诊断学]