hnRNPD在食管鳞状细胞癌的表达及意义

The expression and significance of hnRNPD in esophageal squamous cell carcinoma cells

目的研究食管鳞状细胞癌(ESCC)组织异质性胞核核糖核蛋白D(hnRNPD)的表达与ESCC临床病理特征的关系,通过下调人ESCC细胞中hnRNPD水平,观察对ESCC细胞生长的影响及可能机制。方法用免疫组织化学染色法检测ESCC组织及癌旁组织中hnRNPD的蛋白表达;转染siRNA-hnRNPD下调hnRNPD基因表达,用Western blot法验证沉默效果;MTT法及克隆形成实验检测下调hnRNPD基因表达后ESCC细胞生长情况的变化;采用藻红蛋白标记的膜联素Ⅴ/7-氨基放线菌素D(annexinⅤ-PE/7-AAD)双染色法结合流式细胞术检测基因下调后细胞凋亡变化。结果 hnRNPD在ESCC组织中高表达;hnRNPD在ESCC组织中表达水平与肿瘤分期存在相关性;hnRNPD基因表达的下调能明显抑制ESCC细胞的生长和克隆形成能力;下调hnRNPD基因表达后,与对照组相比,ESCC细胞凋亡明显增加。结论下调hnRNPD基因表达通过增加细胞凋亡抑制ESCC细胞生长及克隆形成能力。

Objective To investigate the expression of heterogeneous nuclear ribonucleoprotein D（ hnRNPD） in esophageal squamous cell carcinoma（ ESCC） tissues and the relationship between hnRNPD expression and the clinicopathological features of ESCC,and to study the effect of down-regulated hnRNPD on the proliferation of ESCC cells and explore its potential mechanism. Methods The expression of hnRNPD protein in ESCC tissues and the normal paracancerous tissues were detected by immunohistochemistry. The siRNA-hnRNPD was transfected into ESCC cells and the silence effect was verified by Western blotting. MTT assay and clone formation assay were used to evaluate the proliferation of ESCC cells after down-regulation of hnRNPD genes. Cell apoptosis was examined by annexin Ⅴ-phycoerythrin /7-aminoactinomycin D（ annexin Ⅴ-PE/7-AAD） staining and flow cytometry. Results The expression of hnRNPD protein in ESCC tissues was significantly higher than that of the normal paracancerous tissues,and the expression was closely related with neoplasm staging. Down-regulation of hnRNPD inhibited the proliferation and clonality of ESCC cells. Compared with the control group,siRNA targeting hnRNPD significantly promoted cell apoptosis. Conclusion Down-regulation of hnRNPD inhibits the proliferation of ESCC cells by promoting cell apoptosis.