转录因子E2F1表达对前列腺癌细胞侵袭能力的影响

Effect of transcription factor E2F1 expression on the invasion of prostate cancer

目的探讨转录因子E2F1对前列腺癌细胞侵袭能力的影响及相关临床意义。方法通过前列腺癌临床基因芯片微列阵数据分析E2F1基因表达水平与临床预后指标的关系；构建稳定沉默E2F1表达的前列腺癌PC3细胞株；运用蛋白免疫印迹方法检测转染细胞株中E2F1和相关肿瘤侵袭分子标志物蛋白表达水平；按照空白对照组和sh-E2F1实验组进行细胞侵袭实验和细胞划痕实验，检测两组在单位时间内的细胞迁移数量。结果E2F1的mRNA表达水平上调与前列腺癌患者术后生化复发率和总体生存率的降低呈正相关（P=0．047，0．035），而且E2F1表达水平与前列腺癌病理分期、Gleason评分和肿瘤转移密切相关（P〈0．05）；稳定沉默E2F1的前列腺癌PC3细胞构建成功，蛋白免疫印迹结果显示E2F1抑制组的波形蛋白（Vimentin）、CD147、基质金属蛋白酶（MMP）-2、MMP-9蛋白表达量与对照组相比下降，而E-钙黏附蛋白（cadherin）的蛋白表达量上升，差异均有统计学意义（均P〈0．05）；抑制E2F1表达的前列腺癌细胞与对照组相比，侵袭和迁移功能均明显减弱，差异有统计学意义（P〈0．05）。结论E2F1通过多种机制增强前列腺癌细胞的侵袭转移能力，其表达水平与前列腺癌患者的不良预后存在重要关系。

Objective To investigate the effect of transcription factor E2F1 on the invasion of prostate cancer and its clinical significance. Methods A stable inhibition of E2F1 prostate cancer cell line PC3 was established. The E2F1 and relative invasion biomarker protein expression level of the transfected cells was detected by Western blot. The PC3 cells were divided into two groups： the control group and sh-E2F1 group, cell invasion assay and cell scratch test were used to detect the number of cell migration in the experiment time. The relationship between E2FI mRNA expression level and clinical prognostic parameters was analyzed through microarray data of prostate cancer. Results E2F1 inhibited PC3 cell line was constructed successfully. The results of Western blot showed that the expression of Vimentin, CD147, MMP-2 and MMP-9 protein in E2F1 suppression group was lower than those in control group, while the protein expression of E-cadherin increased. Compared to the control group, inhibiting the expression of E2F1 in prostate cancer cells significantly decreased the invasion and migration, with significant difference （P 〈 0. 05 ）. High mRNA expression of E2F1 decreased biochemical recurrence rate and overall survival rate after surgery （ P = 0. 047, 0. 035 ） , and the E2F1 expression level was related to pathological stage of prostate cancer. Gleason score and tumor metastasis （ P 〈 0. 05 ）. Conclusion E2F1 enhances the invasion and metastasis of prostate cancer through a variety of mechanisms, and its expression level has an important relationship with the adverse prognosis of patients with prostate cancer.