| 张龙,王瑞,赵佳.肿瘤蛋白翻译控制 1的反义 RNA 1在肺癌中的表达及其对肺癌细胞增殖、迁移和侵袭的影响[J].安徽医药,2022,26(4):718-723. |
| 肿瘤蛋白翻译控制 1的反义 RNA 1在肺癌中的表达及其对肺癌细胞增殖、迁移和侵袭的影响 |
| Expression of TPT1-AS1 in lung cancer and its effect and mechanism on the proliferation, migration and invasion of lung cancer cells |
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| DOI：10.3969/j.issn.1009-6469.2022.04.018 |
| 中文关键词: 肺肿瘤 肿瘤蛋白翻译控制 1的反义 RNA 1 微小 RNA-671-5p 细胞增殖 迁移 侵袭 |
| 英文关键词: Lung neoplasms TPT1-AS1 miR-671-5p Cell proliferation Migration Invasion |
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| 中文摘要: |
| 目的探讨肿瘤蛋白翻译控制 1的反义 RNA 1(TPT1-AS1)在肺癌中的表达及其对肺癌细胞增殖、迁移和侵袭的影响和作用机制。方法收集于 2017年1月至 2018年12月在信阳市中心医院心胸外科行手术治疗的 46例肺癌组织和对应的癌旁组织，实时荧光定量 PCR(RT-qPCR)检测组织中 TPT1-AS1和微小 RNA-671-5p(miR-671-5p)表达， Pearson相关性分析肺癌组织中 TPT1-AS1和miR671-5p表达的相关性。同时，于2019年5月至 2020年4月期间，通过体外培养肺癌细胞 A549，双荧光素酶报告基因实验验证了细胞中 TPT1-AS1和 miR-671-5p调控关系。另将 A549细胞分为对照组、小干扰 RNA阴性序列(si-NC)组、 TPT1-AS1小干扰 RNA(si-TPT1AS1)组、 si-TPT1-AS1+抑制剂阴性序列(anti-miR-NC)组和 si-TPT1-AS1+miR-671-5p抑制剂(anti-miR-671-5p)组，甲基噻唑蓝染色法(MTT)检测细胞增殖， Transwell检测细胞迁移和侵袭，蛋白质印迹实验检测细胞周期蛋白 D1(CyclinD1)基质金属蛋白酶(MMP)-2和 MMP-9蛋白表达。结果肺癌组织中 TPT1-AS1表达量较癌旁组织显著升高［(1.88±0.12)(1.01±0.08)、P<0.05］，miR-671-5p表达量较癌旁组织显著降低［(0.45±0.04)(1.01±0.06)P<0.05］。Pearson相关性分析显示，肺癌组比织中TPT1-A，S1和 miR-671-5p呈负相(r= 0.63，P<0.05)。TPT1-AS1在A5比49细胞中负调，控 miR-671-5p表达。 si-TPT1-AS1组 A549细胞存活率、迁移数和侵袭数分别为(关53.24±2.81)%、(63.11±6.51)个和( 33.78±3.23)个，均低于 si-NC组［分别为( 98.78±2.57)%、(147.44±11.08)个和(101.67±9.95)个，均P<0.05］。CyclinD1、MMP-2和MMP-9蛋白在 si-TPT1-AS1组的表达量分别为(0.43±0.05)(0.21±0.03)(0.43±0.04)，较si-NC组均降低［分别为(0.86±0.04)(0.55±0.05)(0.48±0.07)均P<0.05］。si-TPT1-AS1+anti-miR-671-、5p组A549细、胞存活率、迁移数和侵袭数及 CyclinD1、MMP-2和M、MP-9蛋白表、达分别为(8，5.43±2.94)%、(125.78±10.59)个、(88.78±7.19)个、(0.75±0.03)、(0.48±0.03)、(0.43±0.04)，较 si-TPT1-AS1+anti-miR-NC组均升高［分别为(54.36±2.95)%、(63.78±6.48)个、(33.56±3.54)个、(0.41±0.03)、(0.22±0.03)、(0.21±0.04)，均P<0.05］。对照组与 si-NC组、 si-TPT1-AS1组与 si-TPT1-AS1+anti-miR-NC组各检测指标比较均差异无统计学意义(P>0.05)。结论肺癌组织中 TPT1-AS1表达升高， miR-671-5p表达降低，且两者呈负相关。 TPT1-AS1可能通过靶向下调 miR-671-5p的表达促进肺癌细胞增殖、迁移和侵袭。 |
| 英文摘要: |
| Objective To investigate the expression of tumor protein translation control antisense RNA 1 (TPT1-AS1) in lung cancer and its effect and mechanism on the proliferation, migration and invasion of lung cancer cells.Methods Forty-six cases of lung cancertissues and corresponding adjacent tissues were collected from January 2017 to December 2018 in the Department of CardiothoracicSurgery of Xinyang Central Hospital, and real-time quantitative PCR (RT-qPCR) was used to detect TPT1-AS1 and microRNA-671-5p (miR-671-5p) expression. Pearson correlation was used to analyze the correlation of TPT1-AS1 and miR-671-5p expression in lung can cer tissues. From May 2019 to April 2020, the regulatory relationship between TPT1-AS1 and miR-671-5p in the cells was verified by culturing lung cancer cells A549 in vitro and the dual-luciferase reporter gene assay. A549 cells were divided into the control group, small interfering RNA negative sequence (si-NC) group, TPT1-AS1 small interfering RNA (si-TPT1-AS1) group, si-TPT1-AS1+inhibi tor negative sequence (anti-miR-NC) group and si-TPT1-AS1+miR-671-5p inhibitor (anti-miR-671-5p) group. Cell proliferation was de tected by methylthiazoletrazolium (MTT), cell migration and invasion were detected by Transwell assays, and the protein expression lev els of cyclin D1, matrix metalloproteinase (MMP)-2 and MMP-9 were detected by Western blotting.Results The expression of TPT1AS1 in lung cancer tissue was significantly higher than that in adjacent tissues [(1.88±0.12) vs. (1.01±0.08), P < 0.05], and the expres sion of miR-671-5p was significantly lower than that in adjacent tissues [(0.45±0.04) vs. (1.01±0.06), P < 0.05]. Pearson correlation analysis showed that TPT1-AS1 and miR-671-5p were negatively correlated in lung cancer tissues (r= 0.629, P < 0.05). TPT1-AS1 negatively regulated miR-671-5p expression in A549 cells. The survival rate, migration number and invasion number of A549 cells in the si-TPT1-AS1 group were (53.24±2.81)%, (63.11±6.51)% and (33.78±3.23)%, respectively, which were lower than those in the si-NC group [(98.78±2.57)%, (147.44±11.08)% and (101.67±9.95)%, respectively, P < 0.05]. The protein levels of Cyclin D1, MMP-2 and MMP-9 in the si-TPT1-AS1 group were 0.43±0.05, 0.21±0.03, and 0.43±0.04, respectively, which were lower than those in the si-NC group [(0.86±0.04), (0.55±0.05), and (0.48±0.07), respectively, P < 0.05]. The survival rate, migration number, invasion number, and protein expression of cyclin D1, MMP-2 and MMP-9 in A549 cells in si-TPT1-AS1+anti-miR-671-5p group were (85.43±2.94) %,(125.78±10.59), (88.78±7.19), (0.75±0.03), (0.48±0.03), and (0.43±0.04), respectively, which were higher than those in si-TPT1-AS1+ anti-miR-NC group [(54.36±2.95)%, (63.78±6.48), (33.56±3.54), (0.41±0.03), (0.22±0.03), (0.21±0.04), respectively, P < 0.05]. There was no significant difference between the control group and the si-NC group or between the si-TPT1-AS1 group and the si-TPT1-AS1+ anti-miR-NC group in each detection index (P > 0.05).Conclusions The expression of TPT1-AS1 was increased and the expression of miR-671-5p was decreased in lung cancer tissues, and the two were negatively correlated. TPT1-AS1 may promote lung cancer cell pro liferation, migration and invasion through targeted downregulation of miR-671-5p expression. |
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