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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 study the relationship between the expression of hnRNPD and the clinicopathological features of esophageal squamous cell carcinoma in esophageal squamous cell carcinoma (ESCC), and to observe the effect of hnRNPD on the growth of ESCC cells by down-regulating hnRNPD in human esophageal squamous cell carcinoma (ESCC) Possible mechanism. Methods The protein expression of hnRNPD in ESCC tissues and adjacent normal tissues was detected by immunohistochemistry. The expression of hnRNPD gene was down-regulated by siRNA-hnRNPD, and the silencing effect was verified by Western blot. The expression of hnRNPD gene was detected by MTT assay and clonogenic assay ESCC cells were detected by flow cytometry. Apoptosis was detected by double staining of annexinⅤ-PE / 7-AAD (phycoerythrin labeled with phycoerythrin D) and flow cytometry Variety. Results hnRNPD was highly expressed in ESCC tissues. The expression level of hnRNPD in ESCC tissues was correlated with tumor staging. The down-regulation of hnRNPD gene expression could significantly inhibit the growth and clonality of ESCC cells. After down-regulating hnRNPD gene expression, compared with the control group , ESCC apoptosis increased significantly. Conclusion Down-regulation of hnRNPD gene expression can inhibit ESCC cell growth and clone formation by increasing apoptosis.