WISP1在食管鳞癌组织和细胞中的表达及对细胞侵袭迁移的影响

Expression of WISP1 in tissues and cells of esophageal squamous cell carcinoma and its influence on cell invasion and migration

目的探讨Wnt诱导的分泌型蛋白1(WISP1)在食管鳞癌(ESCC)组织和细胞中的表达及其对ESCC细胞增殖和侵袭迁移的影响。方法应用基因芯片技术筛选ESCC的差异表达基因,免疫组化检测43例ESCC组织的WISP1蛋白水平,实时定量PCR(qPCR)检测78例ESCC组织及人食管上皮细胞HET-1A和ESCC细胞(TE-1、KYSE150、Eca109、KYSE450和EC9706)的WISP1 mRNA水平;脂质体法将靶向WISP1的小干扰RNA(si-WISP1)和阴性siRNA(si-NC)转染Eca109和KYSE450细胞,采用MTT比色法检测增殖活力,Transwell小室实验检测迁移和侵袭能力。结果按照差异倍数≥2.0、P≤0.05的标准在ESCC中筛选出108个差异有统计学意义的基因,其中高表达61个、低表达47个,上调倍数最高的基因依次为WISP1、HOXC8和HMGA2,而下调最高的基因依次为HLF、PLA2G2A和LOC729264。ESCC组织的W1SP1高表达率为72.09%(31/43),高于癌旁组织的32.56%(14/43),ESCC组织中WISP1 mRNA水平为1.882±1.551,高于癌旁组织的-1.784±1.383(P<0.05),分层分析发现有淋巴结转移者的WISP1 mRNA水平高于无淋巴结转移者(P<0.05)。与HET-1A细胞相比,ESCC细胞的WISP1水平升高(P<0.05),而Eca109和KYSE450细胞转染si-WISP1后的WISP1水平、增殖活力和穿膜细胞数均低于转染si-NC的细胞(P<0.05)。结论WISP1在ESCC组织和细胞中均为高表达且参与了ESCC转移过程,下调WISP1可抑制ESCC细胞的增殖和侵袭迁移能力,有望成为ESCC诊治的潜在新靶标。

Objective To investigate the expression of Wnt1 inducible signaling pathway protein 1(WISP1)in tissues and cells of esophageal squamous cell carcinoma(ESCC)and its influence on cell invasion and migration.Methods Differential expression profiles of genes in ESCC were screened by microarray assay.Immunohistochemistry was performed to measure the protein levels of WISP1 in 43 ESCC tissues.The mRNA levels of WISP1 were detected by quantitative polymerase chain reaction(qPCR)in 78 ESCC tissues along with human esophageal epithelial HET-1A cells and ESCC cells including TE-1,KYSE150,Eca109,KYSE450 and EC9706 cells.Small interfering RNA targeting WISP1(si-WISP1)and negative siRNA(si-NC)were transfected into Eca109 and KYSE450 cells by liposome method.The proliferation activity was detected by MTT colorimetry,and the migration and invasion ability were detected by Transwell chamber test.Results According to the criteria of fold change≥2.0 and P≤0.05,108 genes with statistically significant difference were screened in ESCC,including 61 genes with high expression and 47 genes with low expression.Genes with the highest up-regulation were WISP1,HOXC8 and HMGA2,while the genes with the highest down-regulation were HLF,PLA2G2A and LOC729264.The high expression rate of W1SP1 in ESCC tissues was 72.09%(31/43),higher than 32.56%(14/43)in adjacent tissues(P<0.05).The WISP1 mRNA level in ESCC tissues was 1.882±1.551,higher than-1.784±1.383 in adjacent tissues(P<0.05),and stratified analysis showed that the WISP1 mRNA level in patients with lymph node metastasis was higher than that in patients without lymph node metastasis(P<0.05).Compared with HET-1A cells,the WISP1 level in ESCC cells increased(P<0.05),while the WISP1 level,proliferation activity and number of transmembrane cells in Eca109 and KYSE450 cells transfected with si-WISP1 were lower than those transfected with si-NC(P<0.05).Conclusion WISP1 is highly expressed in ESCC tissues and cells and participates in the process of ESCC metastasis.Down-regulation of WISP1 can inhibit the proliferation,invasion and migration of ESCC cells,which is expected to become a potential new target for the diagnosis and treatment of ESCC.