过表达LMO2蛋白的WPMY-1前列腺基质细胞促进PC-3/BPH-1前列腺上皮细胞增殖与侵袭的实验研究

The LMO2 protein overexpressed WPMY-1 prostatic stromal cells promote proliferation and invasion of PC-3/BPH-1 prostatic epithelial cells

目的 研究前列腺外周带强势基因LMO2对前列腺上皮细胞株PC-3、BPH-1增殖和侵袭能力的影响及临床意义.方法 应用慢病毒过表达载体转染WPMY-1前列腺基质肌成纤维细胞株,试图建立过表达LMO2蛋白的WPMY-1细胞株,将WPMY-1细胞株分为实验组（WPMY-1-LMO2,慢病毒Lenti6.3-LMO2-IRES2-EGFP转染WPMY-1细胞）、阴性对照组（WPMY-1-NC,慢病毒Lenti6.3空载体转染WPMY-1细胞）和未经处理的空白对照组（WPMY-1细胞）.采用实时荧光定量PCR和蛋白质印迹法检测LMO2基因和LMO2蛋白的表达情况.将所建立的前列腺基质细胞与PC-3或BPH-1细胞共培养,利用细胞增殖-毒性检测试剂盒和EdU检测PC-3或BPH-1细胞增殖能力的变化情况,利用基质胶侵袭实验检测PC-3或BPH-1细胞侵袭能力的变化情况.结果 成功建立过表达LMO2蛋白的WPMY-1细胞WPMY-1-LMO2.PC-3或BPH-1与WPMY-1-LMO2共培养后增殖能力提高;实验组培养基培养的PC-3细胞EdU阳性细胞百分数为（42.67±6.03）％,与阴性对照组（29.33±3.51）％比较差异有统计学意义（P＜0.05）;实验组培养的BPH-1细胞EdU阳性细胞百分数为（35.00±2.52）％,与阴性对照组（23.33±2.52）％比较差异有统计学意义（P＜0.05）.基质胶侵袭实验检测PC-3或BPH-1与WPMY-1-LMO2共培养后细胞迁移数分别为（38±5）个、（43±3）个,高于对照组,与对照组比较差异均有统计学意义（P＜0.05）.结论 LMO2基因在前列腺基质细胞中高表达促进PC-3和BPH-1细胞的增殖和迁移能力;前列腺不同区带基质细胞的差异表达基因可能是导致前列腺疾病带性差异的重要原因之一.

Objective To investigate the effect of LMO2 gene,which is overexpressed in prostate peripheral zone than transitional zone,on proliferation and invasion of prostate cancer PC-3 cell line or normal prostate epithelial BPH-1 cell line.Methods Lentivirus overexpression system was used to establish the LMO2 protein overexpressed prostate WPMY-1 stromal cell line.The LMO2 mRNA and protein expression level of those cells was evaluated by qRT-PCR and western blot.The PC-3 or BPH-1 cells was cocultured with prostate stromal cells and the in vitro proliferation and invasion activity were detected by Cell Counting Kit-8 （CCK-8）,EdU and Matrigel invasive assays.Results The stable LMO2 overexpressed prostate stromal cells WPMY-1-LMO2 was successfully established.The results of CCK-8,EdU experiments indicated the proliferation and invasion activity of PC-3 or BPH-1 cells were both enhanced through cocultured with WPMY-1-LMO2 cells.The proportion of EdU positive cells of PC-3 cultured in WPMY-1-LMO2 supernatant was （42.67 ± 6.03） %,and the difference was significant compared with the control group （29.33 ± 3.51） % （P 〈 0.05）.The BPH-1 culturcd in the supernatant was （35.00 ± 2.52） %,aud the difference was significant compared with the control group （23.33 ± 2.52） % （P 〈 0.05）.The number of invaded PC-3 or BPH-1 cells after co-cultured with WPMY-1-LMO2 cells was 38 ± 5 and 43 ± 3,respectively,which was significantly different compared with the control group （P 〈 0.05）.Conclusions Tbe LMO2 overexpressed prostate stromal cells could induce proliferation and invasion of PC-3 or BPH-1 cells.The propensity of benign prostatic hyperplasia and prostate cancer at different zones may probably be related to distinctive gene expression between peripheral zone and transitional zone derived stromal cells.