| 杨隆良,郭虎林,马瑜.慢病毒载体介导的长链非编码 RNA Rpph1沉默对胆囊癌 GBC-SD细胞增殖、细胞周期和凋亡的影响[J].安徽医药,2021,25(9):1792-1796. |
| 慢病毒载体介导的长链非编码 RNA Rpph1沉默对胆囊癌 GBC-SD细胞增殖、细胞周期和凋亡的影响 |
| Effects of lentiviral vector-mediated lncRNA Rpph1 silencing on proliferation, cell cycle and apoptosis of gallbladder carcinoma GBC-SD cells |
| |
| DOI：10.3969/j.issn.1009-6469.2021.09.022 |
| 中文关键词: 胆囊肿瘤 长链非编码 RNA Rpph1 慢病毒载体 增殖 细胞周期 凋亡 |
| 英文关键词: Gallbladder neoplasms lncRNA Rpph1 Lentiviral vector Proliferation Cell cycle Apoptosis |
| 基金项目: |
|
| 摘要点击次数: 1141 |
| 全文下载次数: 428 |
| 中文摘要: |
| 目的研究长链非编码 RNA(lncRNA)Rpph1对胆囊癌细胞增殖、细胞周期和凋亡的影响和潜在机制。方法以人胆囊上皮细胞 HGBEC为对照，实时荧光定量逆转录聚合酶链反应( qRT-PCR)检测胆囊癌细胞系 GBC-SD、SGC-996和 NOZ中 lncRNA Rpph1的表达。将 GBC-SD细胞分为对照组、 si-con组、 si-lnc Rpph1组。蛋白质印迹法检测周期蛋白依赖性激酶 2(CDK2)、 Ki-67、剪切型胱天蛋白酶 3(Cleaved caspase-3)、剪切型胱天蛋白酶 9(Cleaved caspase-9)、 β连环素( β-catenin)和 cMyc蛋白表达水平，细胞计数试剂盒( CCK-8)法和流式细胞术分别检测细胞增殖、细胞周期和凋亡率。结果与对照相比，胆囊癌细胞系 GBC-SD、SGC-996和 NOZ组细胞中 lncRNA Rpph1含量［(4.89±0.31)、(3.42±0.42)、(3.96±0.45)比( 1.07±0.13)］显著升高( P<0.05)；与 si-con组比较， si-lnc Rpph1组 GBC-SD细胞活力［(0.78±0.08)比( 1.17±0.13)］降低，凋亡率［( 12.89±1.96)%比(4.26±0.65)%］、 G0~G1期细胞比例［( 63.67±2.62)%比( 52.57±1.32)%］升高， CDK2［( 0.22±0.04)比( 0.41±0.05)］、 Ki-67［( 0.31±0.04)比( 0.62±0.04)］、 β-catenin［( 0.32±0.05)比( 0.63±0.05)］和［c-Myc(0.21±0.04)比( 0.41±0.06)］蛋白表达降低( P<0.05)， Cleaved caspase-3［( 0.44±0.05)比( 0.14±0.04)］和 Cleaved caspase-9［( 0.54±0.05)比( 0.17±0.03)］蛋白表达增加( P<0.05)。结论慢病毒载体沉默 lncRNA Rpph1可能通过 Wnt/β-catenin信号通路抑制胆囊癌 GBC-SD细胞增殖，诱导细胞周期停滞和促进细胞凋亡。 lncRNA Rpph1是胆囊癌的潜在分子靶点。 |
| 英文摘要: |
| Objective To investigate the effects of long non-coding RNA (lncRNA) Ribonuclease P RNA component H1 (Rpph1) onthe proliferation, cell cycle, and apoptosis of gallbladder carcinoma cells and the underlying mechanism.Methods Taking the humangallbladder epithelial cell HGBEC as the control, the expression levels of lncRNA Rpph1 in gallbladder carcinoma cell lines SGC-996, GBC-SD and NOZ were detected by quantitative real-time PCR (qRT-PCR). The GBC-SD cells were divided into control group, si-con group and si-lnc Rpph1 group. The expression levels of cyclin dependent kinase 2 (CDK2), Ki-67, cleaved caspase-3, cleaved caspase9, β-catenin and c-Myc proteins were detected by Western blotting. Cell proliferation, cell cycle and apoptosis rate were measured bycell counting kit (CCK-8) assay and flow cytometry, respectively. The relationship between lncRNA Rpph1 was validated by dual-luciferase reporter assay system.Results Compared with the control group, the levels of lncRNA Rpph1 [(4.89±0.31), (3.42±0.42), (3.96± 0.45) vs. (1.07±0.13)] in gallbladder carcinoma cell lines SGC-996, GBC-SD and NOZ were significantly increased, and the difference was statistically significant (P<0.05). Compared with the si-con group, the viability [(0.78±0.08) vs. (1.17±0.13)] of GBC-SD cell in the si-lnc Rpph1 group was reduced, and the cell apoptosis rate [(12.89±1.96)% vs. (4.26±0.65)%] and G0-G1 stage proportion [(63.67± 2.62)% vs. (52.57±1.32)%] were increased, CDK2 [(0.22±0.04) vs. (0.41±0.05)], Ki-67 [(0.31±0.04) vs. (0.62±0.04)], β-catenin [(0.32± 0.05) vs. (0.63±0.05)] and c-Myc [(0.21±0.04) vs. (0.41±0.06)] protein expression were decreased, cleaved caspase-3 [(0.44±0.05) vs. (0.14±0.04)] and cleaved caspase-9 [(0.54±0.05) vs. (0.17±0.03)] protein expression were increased, and the difference was statistically significant (P<0.05).Conclusions The silencing of lncRNA Rpph1 by lentiviral vector may inhibit the proliferation, induce cell cyclearrest and promote apoptosis of GBC-SD cells through Wnt/β-catenin signaling pathway. LncRNA Rpph1 is a potential molecular target for gallbladder carcinoma. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
