摘要目的 观察明确FOXR2对前列腺癌细胞增殖和凋亡的影响,探讨其潜在作用机制.方法 通过实时定量荧光PCR(qRT-PCR)和蛋白质印迹法(Western印迹)检测FOXR2在前列腺癌临床样本中的表达水平.运用CCK8增殖试剂盒和Annexin V-FITC凋亡试剂盒,流式细胞术检测FOXR2敲低前后前列腺癌细胞增殖和凋亡的变化.结合微小RNA (microRNA)芯片结果预测与FOXR2相互作用的miR-152,并通过荧光素酶报告基因检测miR-152对FOXR2基因的靶向性调控作用.利用软件预测,qRT-PCR技术明确HOTAIR与miR-152的相关性.结果 FOXR2在前列腺癌组织中高表达,其mRNA表达水平是癌旁组织的4.9倍(4.94±0.51比1.00±0.24);蛋白水平也明显上调.在PC3细胞系中,特异性敲低FOXR2会抑制细胞的增殖,促进细胞的凋亡.芯片结果及荧光素酶报告基因检测显示,miR-152能够调控FOXR2的表达;FOXR2 3'UTR有两个miR-152的结合位点,且都能调控FOXR2的表达.LNCediting及qRT-PCR技术提示HOTAIR与miR-152表达呈负相关,在前列腺癌中参与调控miR-152的表达水平.结论 HOTAIR通过靶向抑制miR-152上调FOXR2促进前列腺癌细胞增殖并抑制其凋亡.

abstractsObjective To clarify the effect of FOXR2 on the proliferation and apoptosis of prostate cancer cells and to reveal the mechanism.Methods The expression of FOXR2 in clinical samples of prostate cancer were detected by Quantitative Real-time PCR (qRT-PCR) and Western blotting.The CCK8 proliferation kit and the Annexin V-FITC apoptosis kit,flow cytometry were used to detect the proliferation and apoptosis of prostate cancer cells with or without the FOXR2 knockdown.Combined with the results of microRNA chip,we predicted the related miR-152 and detected the relationship between miR-152 and FOXR2 by luciferase reporter gene assay.The correlation between HOTAIR and miR-152 is clearly defined by software prediction and qRT-PCR.Results FOXR2 had a relatively high expression in the prostate cancer tissue.The mRNA expression of FOXR2 is 4.9 times that of adjacent tissues,and the protein level was also significantly up-regulated.In the PC3 cell line,the specific knock-down of FOXR2 inhibits the proliferation of cells and promotes cell apoptosis.According to the microRNA chip results and luciferase reporter gene assay,we found miR-152 could regulate the expression of FOXR2;and FOXR2 3'UTR had two miR-152 binding sites,all of which could control the expression of FOXR2.The results of LNCediting and qRT-PCR suggest that HOTAIR is negatively correlated with the expression of miR-152,and is involved in the regulation of miR-152 expression in prostate cancer.Conclusion FOXR2 up-regulation can promote the proliferation and inhibit the apoptosis of prostate cancer cells because that HOTAIR restrains the expression of miR-152.