摘要
目的:研究长链非编码RNA(lncRNA) RP1-90L14.1对前列腺癌LNCaP细胞增殖、迁移、侵袭的作用以及对GRIN2A、BACE2基因表达的影响。方法:采用RT-PCR检测前列腺癌LNCaP、LNCaP-AI细胞中RP1-90L14.1的表达水平;在LNCaP细胞中瞬时转染RP1-90L14.1过表达质粒和载体质粒,即转染RP1-90L14.1实验组(LNCaP-RP1-90L14.1组)和转染阴性对照组(LNCaP-NC组);采用普通培养液和活性炭吸附无酚红培养液培养两组细胞;CCK-8法、Transwell法检测两组细胞增殖、迁移、侵袭能力;RT-PCR和Western印迹检测两组细胞中N-甲基-D-天氡氨酸离子型谷氨酸受体2A(GRIN2A)、β位点淀粉样前体蛋白裂解酶2 (BACE2) mRNA和蛋白表达含量变化。结果:RP1-90L14.1在LNCaP-AI中的表达量显著高于LNCaP细胞(8.49±0.43 vs 2.53±0.95,P<0.05)。转染RP1-90L14.1后,LNCaP-RP1-90L14.1组RP1-90L14.1表达量显著高于LNCaP-NC组(0.71±0.22 vs 0.02±0.01,P<0.05);在普通培养液和活性炭吸附无酚红培养液中,培养72 h时,LNCaP-RP1-90L14.1组细胞活性分别高出LNCaP-NC组51.95%(1.22±0.08 vs 0.08±0.05,P<0.05)和50.69%(0.79±0.02 vs 0.53±0.05,P<0.05);培养96 h时,LNCaP-RP1-90L14.1组分别高出LNCaP-NC组51.72%(1.72±0.07 vs 1.13±0.05,P<0.05)和60.23%(1.18±0.05 vs 0.73±0.08,P<0.05)。转染RP1-90L14.1后,在普通培养液和活性炭吸附无酚红培养液中,细胞迁移能力LNCaP-RP1-90L14.1组均显著高于LNCaP-NC组[(682.0±42.7)个vs(422.0±37.1)个,(419.0±42.9)个vs(251.0±25.9)个,P<0.05];细胞侵袭能力LNCaP-RP1-90L14.1组也均显著高于LNCaP-NC组[(507.0±22.2)个vs(274.0±19.6)个,(352.0±14.1)个vs(216.0±14.3)个,P<0.05]。LNCaP-RP1-90L14.1组与LNCaP-NC组相比,GRIN2A mRNA和蛋白表达量(5.13±0.89、2.09±0.54,5.88±0.29、2.03±0.22),BACE2 mRNA和蛋白表达量(5.82±0.50、2.53±0.30,4.89±0.19、3.37±0.13)均有统计学差异(P<0.05)。结论:lncRNA RP1-90L14.1可能在调控前列腺癌细胞的增殖、迁移及侵袭行为中起重要作用;RP1-90L14.1能促进GRIN2A、
Objective: To investigate the effects of long non-coding RNA RP1-90 L14.1 on the proliferation, migration and invasion of prostate cancer LNCaP cells and the expressions of GRIN2A and BACE2. Methods: Using RT-PCR, we detected the expression of RP1-90 L14.1 in LNCaP and LNCaP-AI cells, transiently transfected the RP1-90 L14.1 overexpression plasmid(the RP1-90 L14.1 group) and vector plasmid(the LNCaP-NC group) into the LNCaP cells, and cultured the two groups of cells with ordinary medium and phenol red-free activated carbon adsorption medium(PRF-ACA). Then we examined the proliferation, migration and invasiveness of the cells by CCK-8 and Transwell, and determined the mRNA and protein expressions of GRIN2 A and BACE2 by RT-PCR and Western blot. Results: The expression of RP1-90 L14.1 was significantly higher in the LNCaP-AI than in the LNCaP cells(8.49 ± 0.43 vs 2.53 ± 0.95, P < 0.05), and so was that of LNCaP-RP1-90 L14.1 in the RP1-90 L14.1 than in the LNCaP-NC group after transfection(0.71 ± 0.22 vs 0.02 ± 0.01, P < 0.05). The optical densities(OD) of the cells were 51.95% and 50.69% higher in the RP1-90 L14.1 than in the LNCaP-NC group after 72 hours of culture with ordinary medium and phenol red-free ACA(1.22 ± 0.08 vs 0.08 ± 0.05, P < 0.05;0.79 ± 0.02 vs 0.53 ± 0.05, P < 0.05), and 51.72% and 60.23% higher in the former than in the latter after 96 hours(1.72 ± 0.07 vs 1.13 ± 0.05, P < 0.05;1.18 ± 0.05 vs 0.73 ± 0.08, P < 0.05). The numbers of the migrating cells cultured with common medium and PRF-ACA were markedly higher in the RP1-90 L14.1 than in the LNCaP-NC group after transfection(682.0 ± 42.7 vs 422.0 ± 37.1, P < 0.05;419.0 ± 42.9 vs 251.0 ± 25.9, P < 0.05), and so were those of the invading cells(507.0 ± 22.2 vs 274.0 ± 19.6, P < 0.05;352.0 ± 14.1 vs 216.0 ± 14.3, P < 0.05). Statistically significant differences were observed between the RP1-90 L14.1 and LNCaP-NC groups in the mRNA and protein expressions of GRIN2 A(5.13 ± 0.89 vs 2.09 ± 0.54, P < 0.05;5.88 ± 0.29 vs 2.03 ± 0
作者
吴品庚
张宇曦
张哲
孔垂泽
WU Pin-geng;ZHANG Yu-xi;ZHANG Zhe;KONG Chui-ze(Department of Urology, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 1100()1 , China)
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2019年第3期209-215,共7页
National Journal of Andrology
基金
国家自然科学基金(81001143)
沈阳科学技术计划(17-230-9-18)~~
