| 郑留昌,崔发财,郑培明,等.长链非编码 RNA OPA相互作用蛋白 5反向转录序列 1靶向调控微 RNA-128-3p对结直肠癌细胞增殖、侵袭和转移的影响[J].安徽医药,2024,28(2):259-265. |
| 长链非编码 RNA OPA相互作用蛋白 5反向转录序列 1靶向调控微 RNA-128-3p对结直肠癌细胞增殖、侵袭和转移的影响 |
| The molecular mechanism of long non-coding RNA OIP5-AS1 targeting miR-128-3p to regulate the proliferation, invasion and metastasis of colorectal cancer cells |
| |
| DOI：10.3969/j.issn.1009-6469.2024.02.011 |
| 中文关键词: 结直肠肿瘤 侵袭 转移 微 RNA-128-3p 长链非编码 RNA Opa相互作用蛋白 5-反向 RNA1 |
| 英文关键词: Colorectal neoplasms Invasion Metastasis MiR-128-3p LncRNA OIP5-AS1 |
| 基金项目:国家自然科学基金项目( 81802094) |
|
| 摘要点击次数: 426 |
| 全文下载次数: 230 |
| 中文摘要: |
| 目的探究长链非编码 RNA OPA相互作用蛋白 5反向转录序列 1(lncRNA OIP5-AS1)在结直肠癌中的表达情况及靶向调控微 RNA-128-3p(miR-128-3p)对结直肠癌细胞增殖、侵袭和转移的影响。方法采用实时荧光定量 PCR(qPCR)定量分析 2017年 1月至 2018年 12月在郑州大学人民医院住院并进行手术治疗的 38例结直肠癌病人癌组织及相应癌旁组织、结直肠癌细胞系( SW480、SW620、HT-29和 LoVo)及人正常结直肠黏膜细胞 FHC中 lncRNA OIP5-AS1和 miR-128-3p的表达。将 SW620细胞设为 si-OIP5-AS1组、 si-NC组、 miR-128-3p mimic组、 mimic-NC组、 miR-128-3p inhibitor+si-OIP5-AS1组和 inhibitor-NC+si-OIP5-AS1组，采用细胞计数试剂盒( CCK-8)实验和克隆形成实验检测细胞增殖能力，采用划痕实验和 transwell实验检测细胞侵袭与迁移能力，采用蛋白质印迹法检测 E2F1、细胞周期蛋白 D1(Cyclin D1)、波形蛋白( Vimentin)、 N-钙黏蛋白( N-cadherin)和 E-钙黏蛋白( E-cadherin)蛋白表达，采用双萤光素酶报告实验验证 lncRNA OIP5-AS1与 miR-128-3p的靶向关系。结果癌旁组织相比较， lncRNA OIP5-AS1在结直肠癌组织中表达显著升高( 1.42±0.40比 0.98±0.29，P<0.05)miR-128-3p表达显著降与(0.72±0.21比 1.52±0.39，P<0.05)，且两者在结直肠癌组织中的表达呈负相关；与 FHC细胞( 1.05±0.10相比较， lncRNA OIP5-)，AS低1在结直肠癌细胞株［SW480(1.60±0.25)、 SW620(2.19±0.33)、 HT-29(1.62±0.23)和 LoVo(1.95±0.20)］中的表达显著增高(均 P<0.05)，miR-128-3p表达( 0.70±0.15、0.46±0.03、0.59±0.14、0.74±0.09比 1.02±0.11)显著降低(均 P<0.05)。 lncRNA OIP5-AS1与 miR-128-3p为相互作用靶基因， lncRNA OIP5-AS1表达下调或 miR-128-3p表达上调均能降低 vimentin、N-cadherin、E2F1和 cyclin D1蛋白表达，增高 E-cadherin蛋白表达，抑制 SW620细胞的增殖、侵袭和迁移。在转染 si-OIP5-AS1的同时转染 miR-128-3p inhibitor可增加 vimentin、N-cadherin、E2F1和 cyclin D1蛋白表达，降低 E-cadherin蛋白的表达，逆转 lncRNA OIP5-AS1表达下调对 SW620细胞增殖、侵袭和迁移的抑制作用。结论 lncRNA OIP5-AS1在结直肠癌中高表达，并靶向调控 miR-128-3p参与结直肠癌的增殖、侵袭和迁移过程， lncRNA OIP5-AS1和 miR-128-3p能为结直肠癌的诊断和靶向治疗提供潜在的应用价值。 |
| 英文摘要: |
| Objective To investigate the expression of long non-coding RNA (lncRNA) Opa interacting protein 5 antisense RNA l (OIP5-AS1) in colorectal cancer (CRC) and its effect on the proliferation, invasion and metastasis of CRC cells by targeting miR-128-3p. Methods The expressions of lncRNA OIP5-AS1 and miR-128-3p in cancer tissues and adjacent normal tissues of patients admittedto The People's Hospital of Zhengzhou University and undergoing surgery from January 2017 to December 2018, CRC cell lines(SW480, SW620, HT-29 and LoVo) and human normal colorectal mucosal cells FHC were quantitatively analyzed by qPCR. SW620cells were transfected with si-OIP5-AS1, si-NC, miR-128-3p mimics, mimics-NC, miR-128-3p inhibitor+si-OIP5-AS1 or inhibitor-NC+ si-OIP5-AS1, the proliferation, invasion and migration of SW620 cells were detected by clone formation assay, CCK-8 assay, wound-healing assay and transwell assay, respectively. The expressions of E2F1, cyclin D1, Vimentin, N-cadherin and E-cadherin were detect. ed by Western blotting assay. Dual luciferase reporter assay was used to verify the targeting relationship between lncRNA OIP5-AS1 and miR-128-3p.Results Compared with adjacent normal tissues, the expression of lncRNA OIP5-AS1 in CRC tissues was signifi. cantly increased (1.42±0.40 vs. 0.98±0.29, P<0.05), and the expression of miR-128-3p was significantly decreased (0.72±0.21 vs. 1.52± 0.39, P<0.05), and the expressions of lncRNA OIP5-AS1 and miR-128-3p in CRC tissues were negatively correlated. Compared with FHC cells, the expressions of lncRNA OIP5-AS1 in CRC cell lines [(1.05±0.10) vs. SW480 (1.60±0.25), SW620 (2.19±0.33), HT-29 (1.62±0.23) and LoVo (1.95±0.20)] were significantly increased, and the expressions of miR-128-3p were significantly decreased [(0.70±0.15), (0.46±0.03), (0.59±0.14), (0.74±0.09) vs. (1.02±0.11)]. LncRNA OIP5-AS1 and miR-128-3p were target genes interacting with each other. Down-regulation of lncRNA OIP5-AS1 or up-regulation of miR-128-3p could decrease the expressions of Vimentin, N-cadherin, E2F1 and cyclin D1, increase the expression of E-cadherin, and inhibit the proliferation, invasion and migration of SW620 cells. Co-transfection of si-OIP5-AS1 and miR-128-3p inhibitor could increase the expressions of Vimentin, N-cadherin, E2F1 and cy. clin D1, decrease the expression of E-cadherin, and reverse the inhibitory effect of down-regulation of lncRNA OIP5-AS1 on the prolif. eration, invasion and migration of SW620 cells.Conclusions LncRNA OIP5-AS1 is highly expressed in CRC and is involved in the proliferation, invasion and migration of CRC by targeting miR-128-3p. LncRNA OIP5-AS1 and miR-128-3p could provide potential ap. plication value for the diagnosis and targeted therapy of colorectal cancer. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
