RNA聚合酶2延长因子相关因子2通过调节骨髓基质抗原2的表达调控前列腺癌细胞的增殖与迁移

Bone marrow stromal antigen 2 , a novel ELL - associated factor 2 target gene, regulates prolifera- tion and migration abilities of prostate cancer cells

目的 观察雄激素应答基因RNA聚合酶2延长因子相关因子2（EAF2）对骨髓基质抗原2（BST2）的调节能力，以及EAF2通过BST2调节前列腺癌细胞增殖和迁移的能力。方法 以Lipofectamine 2000试剂转染针对EAF2和非特异性对照组的小干扰RNA（siRNA，分别为100 pmol），干扰前列腺癌细胞C4-2中EAF2的表达，采用实时定量聚合酶链反应（Real-time PCR）和Western blot检测细胞内BST2的mRNA和蛋白水平表达变化；采用质粒转染的方法在293T细胞中过表达EAF2后检测BST2蛋白水平的变化，明确EAF2对BST2表达的影响；采用溴脱氧尿嘧啶核苷（BrdU）标记法和Transwell小室法计算并检测干扰BST2表达后对前列腺癌细胞C4-2增殖能力和迁移能力的影响，明确BST2对前列腺癌细胞增殖和迁移的调节能力。结果 干扰EAF2表达后细胞内BST2 mRNA的表达明显升高，说明EAF2可以调节BST2的转录水平，进一步检测干扰EAF2表达后，BST2的蛋白水平表达明显升高，进一步干扰BST2表达后，检测到前列腺癌细胞的增殖比例由（22.09±1.08）%下降至（12.25±1.32）%（P＝0.001）。同时，迁移细胞的数量从（50.50±3.80）%下降至（24.25±1.42）%（P＝0.001）。结论 在前列腺癌细胞中，EAF2可以在转录和蛋白水平调节BST2的表达，BST2具有调节前列腺癌细胞增殖和迁移的能力。

Objective To investigate whether bone marrow stromal antigen 2 （BST2） which was regulated by ELL-associated factor 2 （EAF2） can promote prostate cancer cells proliferation and migration.Methods After inhibiting the expression of EAF2 by 100 pmol interfering RNA （RNAi） in C4-2 cells using lipofectamine 2000, the mRNA and protein expression levels were assessed by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting respectively. After transfecting MYC-EAF2 into 293T cells according to the manufacturer’s instruction, the expression level of BST2 was detected by Western blotting. After knockdown of EAF2 by RNAi in C4-2 cells, proliferation was tested by bromodeoxyuridine incorporation （BrdU） assay, and migration was tested by Transwell assay.Results The mRNA and protein levels were significantly increased after transfection of small interfering RNA （siRNA） against EAF2. Moreover, proliferation rate was further decreased from （22.09±1.08）% to （12.25±1.32）% （P=0.001）. At the meantime, the number of migrating cells was decreased from （50.50±3.80）% to （24.25±1.43）% （P=0.001）.Conclusion Our findings suggest that EAF2 regulates the mRNA and protein levels of BST2, and BST2 could regulate proliferation and migration abilities of C4-2 cells.