子宫颈鳞状细胞癌中DIAPH3表达对细胞迁移和侵袭的作用及分子机制

Effect of DIAPH3 on migration and invasion of cervical squamous cell carcinoma cell and its molecular mechanism

目的探讨Diaphanous相关成蛋白3(diaphanous-related formin 3,DIAPH3)对子宫颈鳞状细胞癌(cervical squamous cell carcinoma,CSCC)细胞迁移和侵袭的作用及其分子机制。方法收集30例CSCC石蜡包埋组织及配对癌旁组织,应用生物信息学与免疫组化分析CSCC组织中DIAPH3的表达,并进行临床病理相关性分析。采用分子生物学与肿瘤迁移相关实验分析DIAPH3对CSCC细胞迁移和侵袭的作用及机制。结果仙桃学术数据库分析显示:DIAPH3 mRNA在CSCC组织(2.37)中表达比癌旁组织(0.25)明显升高(P<0.05);DIAPH3高表达患者的无复发生存率明显降低(P=0.031)。免疫表型:CSCC组织中DIAPH3的阳性率为70.00%(21/30),高于癌旁组织(30%,9/30);DIAPH3的阳性率在不同肿瘤分化程度(P=0.032)和有无淋巴结转移(P=0.018)组间差异有统计学意义。siRNA干扰DIAPH3组:CSCC细胞的迁移率(28.33%,P=0.002)、穿过小室的细胞数(207.67,P=0.001)及p-ERK(P=0.001)、MMP-2(P=0.003)、MMP-9(P=0.002)蛋白表达量均明显降低。过表达DIAPH3组:CSCC细胞的迁移率(79.00%,P=0.0004)和穿过小室的细胞数(224.33,P=0.0003)及p-ERK(P=0.001)、MMP-2(P=0.001)、MMP-9(P=0.002)蛋白表达量均明显升高。DIAPH3+U0126组:CSCC细胞的迁移率(46.00%,P=0.0001)和穿过小室的细胞数(106.33,P=0.0003)及p-ERK(P=0.001)、MMP-2(P=0.002)、MMP-9(P=0.004)蛋白表达量均明显降低。结论DIAPH3在CSCC组织中高表达,与肿瘤分化程度和淋巴结转移密切相关,且通过ERK1/2通路促进CSCC细胞的迁移和侵袭。

Purpose To investigate the effect and molecular mechanism of diaphanous-related formin 3(DIAPH3)on the migration and invasion of cervical squamous cell carcinoma cell.Methods The paraffin-embedded tissue and matched paracancerous tissue samples of 30 patients with cervical squamous cell carcinoma were collected,and the expression of DIAPH3 in cervical squamous cell carcinoma tissues was analyzed by bioinformatics and immunohistochemistry,and the clinicopathological correlation was analyzed.The effect of DIAPH3 on migration and invasion of cervical squamous cell carcinoma cells and its mechanisms were analyzed by molecular biology and tumor migration related experiments.Results The expression of DIAPH3 mRNA in CSCC tissue(2.37)was significantly higher than that in adjacent tissues(0.25)(P<0.05),the recurrence free survival of patients with high expression of DIAPH3 was significantly reduced(P=0.031).Immunophenotyping showed that the positive rate of DIAPH3 in cervical squamous cell carcinoma was 70.00%(21/30),which was higher than that in paracancerous tissues(30%,9/30).According to the degree of tumor differentiation and lymph node metastasis,the positive rate of DIAPH3 was significantly different(P=0.032,P=0.018).After siRNA interference with DIAPH3,the migration rate(28.33%,P=0.002),the number of cells through the chamber(207.67,P=0.001)and the expression of p-ERK(P=0.001),MMP-2(P=0.003),MMP-9(P=0.002)protein were significantly reduced.After overexpression of DIAPH3,the migration rate(79.00%,P=0.0004),the number of cells through the chamber(224.33,P=0.0003)and the expression of p-ERK(P=0.001),MMP-2(P=0.001),MMP-9(P=0.002)protein were significantly elevated,while in DIAPH3+U0126 group,the migration rate(46.00%,P=0.0001),the number of cells through the chamber(106.33,P=0.0003)and the expression of p-ERK(P=0.001),MMP-2(P=0.002),MMP-9(P=0.004)protein were significantly reduced.Conclusion DIAPH3 is highly expressed in cervical squamous cell carcinoma,closely related to tumor differentiation and lymph node metastasis,and promotes the migration and invasion of cervical squamous cell carcinoma cells through ERK1/2 pathway.