| 李二毛 陈嘉欣 李晓艳 邹东霆 李冰 刘启才.慢病毒介导的稳定表达LBH的前列腺癌细胞株的建立[J].,2016,16(3):431-434 |
| 慢病毒介导的稳定表达LBH的前列腺癌细胞株的建立 |
| The Establishment of PC-3MCell Stably Expressing LBH Mediated byLentiviral |
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| DOI： |
| 中文关键词: LBH 基因 前列腺癌 PC-3M 细胞增殖 |
| 英文关键词: LBH gene Prostatic carcinoma PC-3M Cell proliferation |
| 基金项目:广东省自然科学基金项目（06301164） |
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| 中文摘要: |
| 目的：构建稳定表达LBH基因的人前列腺癌细胞株PC-3M-LBH，探讨LBH基因对PC-3M 细胞增殖能力的影响。方法：构
建表达LBH基因的重组慢病毒载体并制备出相应的慢病毒，感染低表达LBH 基因的人前列腺癌PC-3M细胞后，经嘌呤霉素筛
选获得细胞克隆；实时荧光定量PCR和蛋白印迹法（Western-Blot）分别检测细胞株中LBH的mRNA、蛋白表达水平；采用CCK-8
法检测表达LBH 基因后细胞增殖能力的改变。结果：成功构建了重组慢病毒表达质粒pLenti-LBH 并包装出了慢病毒，感染前列
腺癌细胞后经嘌呤霉素筛选得到PC-3M-LBH 细胞株；PC-3M-LBH细胞株中LBH基因的mRNA 和蛋白表达显著上调；相对母
细胞和NC对照组，PC-3M-LBH 细胞在接种后第4 天即出现明显的生长抑制，到第6 天其生长抑制率达到19.7 %。结论：构建的
细胞株能稳定表达LBH 基因，该基因的表达能显著抑制前列腺癌PC-3M细胞的体外增殖。 |
| 英文摘要: |
| Objective:To establish a PC-3M cell line stably expressing LBH gene and explore the effect of LBH gene on the proliferation
of PC-3M cells.Methods:Recombinant lentiviral expression vector containing LBH gene was constructed and lentivirus was
produced. After lentivirus infection of human prostatic carcinoma PC-3M cells, PC-3M-LBH cell line was obtained by screening with
puromycin; Expression of LBH in different cells was examined by RT-PCR and Western blot; Cell proliferation was detected by CCK-8
assay.Results:Lentiviral expression plasmid plenti-LBH was constructed and lentivirus particle was successfully packed. After infected
with lentivirus and screened by puromycin, a cell line PC-3M-LBH was obtained. LBH expression in PC-3M-LBH was significantly
higher compared with the NC control group and parental cells. The proliferation of PC-3M-LBH cells decreased 19.7 % compared with
NC control and PC-3Mat the 6th day after cells seeding.Conclusion:LBH gene expression can inhibit the proliferation of prostate cancer
PC-3Mcells. |
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