角鲨烯环氧酶对宫颈鳞癌细胞增殖、凋亡及迁移侵袭的影响

Effects of SQLE gene on proliferation,apoptosis,migration and invasion of squamous cervical cancer cells

目的探究角鲨烯环氧酶(squalene epoxidase,SQLE)在宫颈鳞癌中的作用及其分子机制。方法首先利用GEPIA和HPA数据库分析SQLE在宫颈鳞癌与正常宫颈组织中mRNA及蛋白质表达差异,运用UALCAN数据库探讨SQLE与宫颈鳞癌临床病理特征的相关性,然后通过GEPIA数据库评估高表达SQLE mRNA的宫颈鳞癌患者的OS和DFS,最后以Siha细胞为研究对象,以siRNA抑制SQLE基因的表达及转染重组质粒促进SQLE的表达,观察SQLE基因对Siha细胞增殖、凋亡及迁移侵袭的影响,qRT-PCR法检测SQLE mRNA表达,Western blot检测Bax、Bcl-2、Vimentin、E-cadherin、PI3K、Akt、p-PI3K和p-Akt蛋白水平。结果SQLE mRNA和蛋白水平在宫颈鳞癌组织中较正常组织中高表达(P<0.05),且SQLE mRNA高表达的宫颈鳞癌患者的OS明显缩短(P<0.05)。SQLE在宫颈鳞癌StageⅣ期的表达明显高于Ⅰ、Ⅱ、Ⅲ期(P<0.01);SQLE在淋巴结转移N1组的表达明显高于N0组(P<0.01)。干扰SQLE能够明显抑制Siha细胞增殖、迁移侵袭及促进其凋亡(P<0.01),过表达SQLE时结果与之相反。干扰SQLE时Siha细胞Bcl-2、Vimentin、p-PI3K和p-Akt蛋白表达降低,Bax、E-cadherin蛋白表达增加,且Bcl-2/Bax比值明显减少(P<0.05,P<0.01),过表达SQLE时结果与之相反。结论SQLE在人宫颈鳞癌中高表达,SQLE可能通过调控PI3K/Akt信号通路诱导Siha细胞增殖、迁移侵袭并抑制其凋亡。

Aim To explore the effect of squalene epoxidase(SQLE)in cervical squamous cell carcinoma and the molecular mechanism.Methods Firstly,the gene expression profiling interactive analysis(GEPIA)database was used to analyze the mRNA expression of SQLE in cervical squamous cell carcinoma and normal cervical tissues,and the human protein atlas(HPA)database was used to obtain the expression of SQLE protein in cervical squamous cell carcinoma and normal cervical tissues.We researched the correlation between SQLE gene and the clinicopathological characteristics of cervical squamous cell carcinoma through UALCAN database.Then GEPIA database was utilized to evaluate the overall survival(OS)and disease free survival(DFS)of cervical squamous cell carcinoma patients with high expression of SQLE mRNA.Finally,Siha cells were taken as the research object,and the effects of SQLE gene on proliferation,apoptosis,migration and invasion of Siha cells were observed by using small interfering RNA(siRNA)to inhibit the expression of SQLE gene and transfecting recombinant plasmid to promote the expression of SQLE gene.The mRNA expression of SQLE was assessed by qPT-PCR.Bax,Bcl-2,Vimentin,E-cadherin,PI3K,Akt,p-PI3K and p-Akt protein expression levels were examined by Western blot.Results The mRNA expression and protein expression of SQLE in cervical squamous cell carcinoma was higher than that in normal tissues(P<0.05),and the OS of patients with high expression of SQLE mRNA was significantly shortened in cervical squamous cell carcinoma(P<0.05).The expression of SQLE in stage IV of cervical squamous cell carcinoma was significantly higher than that in stage I,II and III(P<0.01).And the expression of SQLE in lymph node metastasis N1 group was markedly higher than that in N0 group(P<0.01).Cell experiments showed that interference with SQLE could significantly inhibit the proliferation,migration and invasion of Siha cells,and promote their apoptosis(P<0.01).The trend was opposite when SQLE was overexpressed.SQLE knockdown decreased the protein expression levels of Bcl-2,Vimentin,p-PI3K and p-Akt,increased the protein expression levels of Bax and E-cadherin,and the ratio of Bcl-2/Bax decreased significantly(P<0.05,P<0.01).The trend was opposite when SQLE was overexpressed.Conclusions SQLE is highly expressed in human cervical squamous cell carcinoma.SQLE may induce Siha cell proliferation,migration,invasion,and inhibit their apoptosis by regulating PI3K/Akt signaling pathway.