Ezrin在前列腺癌中的功能研究及临床意义

Ezrin in vitro functional and clinical significance research for prostate cancer

目的研究Ezrin蛋白在前列腺癌细胞中的功能及其在高危前列腺癌患者根治术术后生存的预测意义。方法用pc DNA3.1为载体,过表达及干扰Ezrin基因(A:空白组,B:pc DNA3.1-NC组,C:pc DNA3.1-Ezrin组,D:si-pc DNA3.1-Ezrin组和E:si-pc DNA3.1-NC组)。RT-PCR检测22RV1、PC-3细胞及前列腺癌组织中Ezrin基因m RNA表达,迁移和侵袭实验评估Ezrin表达变化后对各组细胞的影响。免疫组化验证高危前列腺癌患者组织中Ezrin表达与术后生化复发、临床进展及总体生存之间的差异。结果对比BPH,Ezrin在前列腺癌细胞22RV1和PC-3表达明显升高[(1.00±0.01)(3.52±0.39)vs(1.00±0.01)(6.27±0.53),P<0.05]。Ezrin基因的m RAN表达与前列腺癌危险分级呈正相关(P<0.05)。A-E组两株细胞处理后Ezrin基因的表达为:22RV1(1.00±0.01、1.12±0.06、1703.27±153.62、67.35±11.49和0.23±0.02);PC-3(0.96±0.01、1.16±0.03、1931.65±183.50、58.71±15.24和0.19±0.02)。结果表明,22RV1和PC-3细胞中,A组与B组差异均无统计学意义(P=0.785、P=0.721);B组与C组、B组与D组、B组与E组、C组与D组差异均有统计学意义(P<0.001)。C组相对B组的迁移和侵袭能力均明显增强(P≤0.001)。免疫组化验证Ezrin蛋白阳性表达率为75.29%(192/255),免疫组化评分3分、4分、5分三组中生化复发(P<0.001)、进展(P=0.015)及总体生存(P=0.036)均有统计学差异。结论过表达Ezrin蛋白使肿瘤细胞的迁移和侵袭能力明显增强,高危列腺癌患者术后5年的生化复发、进展及总体生存与Ezrin表达量呈正相关性。

Objective To explore the function of Ezrin protein in prostate cancer cells and its significance to predict survival in high-risk prostate cancer who had underwent radical prostatectomy.Methods Ezrin gene expression and interference with pc DNA3.1 and si RNA（A：blank group,B：pc DNA3.1-NC group,C：pc DNA3.1-Ezrin group,D：si-pc DNA3.1-Ezrin group and E：si-pc DNA3.1-NC group）.RT-PCR was used to detect m RNA expression of Ezrin in 22RV1,PC-3 and prostate cancer tissues.The effect on each group from the change of Ezrin expression was assessed by migration and invasion experiment.The difference between Ezrin expression and biochemical recurrence,progression,overallsurvival of high-risk postoperatively radical prostatectomy were studied by immunohistochemisty.Results Contrast with BPH cell,Ezrin gene overexpression in 22RV1 cell and PC-3 cell were elevated significantly[（1.00 ±0.01）（3.52±0.39） vs（1.00±0.01）（6.27±0.53）,P0.05].Positively correlated with the expression of Ezrin m RNA in low risk,intermediate risk,high-risk and lymph node metastatic prostate cancer was detected（2.33 ±0.52,2.64 ±0.22,4.58 ±0.39,7.02 ±0.78,P0.05）.Compared Ezrin expression in A-E groups,22RV1（1.00±0.007,1.12±0.062,1703.27±153.622,67.35±11.492 and 0.23±0.019）;PC-3（0.96±0.011,1.16±0.033,1931.65±183.502,58.71±15.244 and 0.19±0.022）.The results that both in 22RV1 and PC-3,there were no significant differences between group A and group B（P=0.785,P=0.721）;Significant differences were found in group B and group C、group B and group D,group B and group E,group C and group D（all P0.001）.Migration and invasion ability of Group C was more stronger than group B（ P ≤ 0.001）.The positive expression of Ezrin by immunohistochemisty was 75.29%（192/255）.Significant differences in biochemical recurrence（P0.001）,progression（P=0.015） and overall survival（P=0.036） were found between the score 3,4,5 Group.Conclusion Our study successfully constructed and verified that overexpression of Ezrin significantly enhanced the migration and invasion ability of 22RV1 cells and PC-3 cells.5 years biochemical recurrence,progression and overall survival of high-risk prostate cancer patients who had underwent radical prostatectomy were validated positively correlated with the expression of Ezrin.