| 45 | 0 | 1340 |
| 下载次数 | 被引频次 | 阅读次数 |
目的 探讨长链非编码RNA(LncRNA)生长停滞特异基因转录的反义RNA(GAS6-AS1)调节微小RNA-708-5p(miR-708-5p)/丙酮酸脱氢酶激酶4(PDK4)轴对喉癌细胞增殖、凋亡和侵袭的影响。方法 实时荧光定量PCR(qRT-PCR)法检测喉癌组织标本中LncRNA GAS6-AS1、miR-708-5p、PDK4的表达；将喉癌TU686细胞分为：siRNA阴性对照组(si-NC组)、单独抑制GAS6-AS1组(si-GAS6-AS1组)、si-GAS6-AS1与抑制剂阴性对照共转染组(si-GAS6-AS1+anti-NC组)以及si-GAS6-AS1与anti-miR-708-5p共转染组(si-GAS6-AS1+anti-miR-708-5p组);检测各组TU686细胞中LncRNA GAS6-AS1、miR-708-5p及PKD4的表达水平；平板克隆法、流式细胞仪、Transwell实验分别检测TU686细胞增殖、凋亡、迁移和侵袭能力；Western blotting法检测蛋白表达；双荧光素酶报告基因实验验证基因的靶向关系。结果 喉癌组织LncRNA GAS6-AS1、PDK4的表达水平显著升高，而miR-708-5p表达水平显著降低(P<0.05)。si-GAS6-AS1组TU686细胞中LncRNA GAS6-AS1表达、PDK4 mRNA表达、克隆细胞数、迁移细胞数、侵袭细胞数及PCNA、MMP-9和PDK4的蛋白表达均低于si-NC组，而miR-708-5p表达、细胞凋亡率及Bax蛋白表达高于si-NC组(P<0.05);与si-GAS6-AS1组、si-GAS6-AS1+anti-NC组比较，si-GAS6-AS1+anti-miR-708-5p组PDK4 mRNA表达、克隆细胞数、迁移细胞数、侵袭细胞数及PCNA、MMP-9和PDK4的蛋白表达水平升高，而miR-708-5p表达、凋亡率及Bax蛋白表达降低(P<0.05)。LncRNA GAS6-AS1靶向负调控miR-708-5p表达，miR-708-5p靶向负调控PDK4表达。结论 抑制LncRNA GAS6-AS1可能通过靶向miR-708-5p来下调PDK4表达，进而抑制TU686细胞增殖、侵袭，促进其凋亡。
Abstract：Objective To investigate the effects of long non-coding RNA(LncRNA) growth arrest-specific gene 6-antisense RNA 1(GAS6-AS1) on the proliferation, apoptosis, and invasion of laryngeal cancer cells by regulating the microRNA-708-5p(miR-708-5p)/pyruvate dehydrogenase kinase isozyme 4(PDK4) axis. Methods The quantitative real-time(qRT-PCR) was applied to detect the expression of LncRNA GAS6-AS1, miR-708-5p, and PDK4 in laryngeal cancer tissue samples. Laryngeal cancer TU686 cells were divided into siRNA negative control(si-NC group), GAS6-AS1 knockdown alone group(si-GAS6-AS1 group), co-transfection with si-GAS6-AS1 and inhibitor negative control group(si-GAS6-AS1+anti-NC group), and co-transfection with si-GAS6-AS1 and anti-miR-708-5p group(si-GAS6-AS1+anti-miR-708-5p group), and the expression level in TU686 cells of each group was detected. The proliferation, apoptosis, migration and invasion of TU686 cells were detected by plate cloning assay, flow cytometry and Transwell assay, respectively. Protein expression was detected by Western blotting method. The dual-luciferase reporter gene assay was used to verify the interaction of genes respectively. Results The expression levels of LncRNA GAS6-AS1 and PDK4 in laryngeal cancer tissue were obviously increased, while the expression level of miR-708-5p was obviously reduced(P<0.05). The expression of LncRNA GAS6-AS1 and PDK4 mRNA, numbers of cloned cells, migrated cells and invasive cells, PCNA, MMP-9, and PDK4 protein expression in TU686 cells in the si-GAS6-AS1 group were lower than those in the si-NC group, and the levels of miR-708-5p, apoptosis rate, and Bax protein expression in the si-GAS6-AS1 group were higher than those in the si-NC group(P<0.05). Compared with the si-GAS6-AS1 group and si-GAS6-AS1+anti-NC group, the expression of PDK4 mRNA, numbers of cloned cells, migrated cells and invasive cells, PCNA, MMP-9, and PDK4 protein expression increased in the si-GAS6-AS1+anti-miR-708-5p group, while the miR-708-5p expression, apoptosis rate, and Bax protein expression decreased(P<0.05). LncRNA GAS6-AS1 targeted and negatively regulated miR-708-5p expression, while miR-708-5p targeted and negatively regulated PDK4 expression. Conclusion Inhibition of LncRNA GAS6-AS1 may downregulate PDK4 expression by targeting miR-708-5p, thereby inhibiting TU686 cell proliferation and invasion, and promoting apoptosis.
[1] Igissin N,Zatonskikh V,Telmanova Z,et al.Laryngeal cancer:epidemiology,etiology,and prevention:A narrative review[J].Iran J Public Health,2023,52(11):2248-2259.
[2] Gu J,Han T,Sun L,et al.miR-552 promotes laryngocarcinoma cells proliferation and metastasis by targeting p53 pathway[J].Cell Cycle,2020,19(9):1012-1021.
[3] Wang Y,Ma M,Li C,et al.GAS6-AS1 overexpression increases GIMAP6 expression and inhibits lung adenocarcinoma progression by sponging miR-24-3p[J].Front Oncol,2021,11(1):645771-645781.
[4] 郑宏，曾振华，唐建生，等.circ-RAD23B通过miR-708-5p/CPNE1轴调控膀胱癌细胞增殖、细胞周期、凋亡的机制[J].中国老年学杂志，2022,42(17):4314-4320.
[5] 吕翠婷，尹情，韩俊淑，等.LINC00665通过靶向miR-708-5p/DKK3抑制食管癌细胞的增殖及侵袭[J].临床与实验病理学杂志，2022,38(4):410-415.
[6] Liu X,Shen X,Zhang J.Long non-coding RNA LINC00514 promotes the proliferation and invasion through the miR-708-5p/HOXB3 axis in cervical squamous cell carcinoma[J].Environ Toxicol,2022,37(1):161-170.
[7] Wu X,Tan Y,Tang X.Long noncoding RNA MALAT1 promotes laryngocarcinoma development by targeting miR-708-5p/BRD4 axis to regulate YAP1-mediated epithelial-mesenchymal transition[J/OL].Biomed Res Int,2022[2024-12-20].https://pubmed.ncbi.nlm.nih.gov/35601153/.
[8] Dou X,Fu Q,Long Q,et al.PDK4-dependent hypercatabolism and lactate production of senescent cells promotes cancer malignancy[J].Nat Metab,2023,5(11):1887-1910.
[9] Xu S,Guo J,Zhang W.lncRNA PCAT19 promotes the proliferation of laryngocarcinoma cells via modulation of the miR-182/PDK4 axis[J].J Cell Biochem,2019,120(8):12810-12821.
[10] Dong J,Wang J,Shan C,et al.MicroRNA-892a regulates laryngocarcinoma cell proliferation via Dicer[J].Exp Biol Med (Maywood),2020,245(14):1222-1232.
[11] Li Z,Liu J,Peng H,et al.SKA1 is overexpressed in laryngocarcinoma and modulates cell growth via P53 signaling pathway[J].Cell Cycle,2023,22(6):693-704.
[12] Zhang P,Dong Q,Zhu H,et al.Long non-coding antisense RNA GAS6-AS1 supports gastric cancer progression via increasing GAS6 expression[J].Gene,2019,696(1):1-9.
[13] Guo X,Li H,Zhang M,et al.LncRNA GAS6 antisense RNA 1 facilitates the tumorigenesis of clear cell renal cell carcinoma by regulating the AMP-activated protein kinase/mTOR signaling pathway[J].Oncol Lett,2021,22(4):727-737.
[14] Chen Q,Zhou L,Ma D,et al.LncRNA GAS6-AS1 facilitates tumorigenesis and metastasis of colorectal cancer by regulating TRIM14 through miR-370-3p/miR-1296-5p and FUS[J].J Transl Med,2022,20(1):356.
[15] Wu XP,Xu Z Q,Xie WM,et al.Long non-coding RNA GAS6-AS1 enhances breast cancer cell aggressiveness by functioning as a competing endogenous RNA of microRNA-215-5p to enhance SOX9 expression[J].Exp Ther Med,2022,23(1):109-115.
[16] Yan Z,Liu G,Liang M,et al.Ophiopogonin D inhibits cell proliferation and induces apoptosis of human laryngocarcinoma through downregulation of cyclin B1 and MMP-9 and upregulation of p38-MAPK signaling[J].Oncol Lett,2019,17(2):1877-1882.
[17] Wu X,Tan Y,Tang X.Long noncoding RNA MALAT1 promotes laryngocarcinoma development by targeting miR-708-5p/BRD4 axis to regulate YAP1-mediated epithelial-mesenchymal transition[J].Biomed Res Int,2022,1(1):1-11.
[18] Wu X,Cui F,Chen Y,et al.Long non-coding RNA LOXL1-AS1 enhances colorectal cancer proliferation,migration and invasion through miR-708-5p/CD44-EGFR Axis[J].Onco Targets Ther,2020,13(1):7615-7627.
[19] Dong HT,Liu Q,Zhao T,et al.Long Non-coding RNA LOXL1-AS1 drives breast cancer invasion and metastasis by antagonizing miR-708-5p expression and activity[J].Mol Ther Nucleic Acids,2020,19(1):696-705.
[20] Pin F,Novinger L J,Huot J R,et al.PDK4 drives metabolic alterations and muscle atrophy in cancer cachexia[J].FASEB J,2019,33(6):7778-7790.
[21] Liu B,Yan L,Chi Y,et al.Long non-coding RNA AFAP1-AS1 facilitates ovarian cancer progression by regulating the miR-107/PDK4 axis[J].J Ovarian Res,2021,14(1):60-70.
基本信息:
中图分类号:R739.65
引用信息:
[1]谢洋,麻琳,要兆旭,等.LncRNA GAS6-AS1调节miR-708-5p/PDK4轴对喉癌细胞增殖、凋亡和侵袭的影响[J].临床肿瘤学杂志,2025,30(08):729-734.
2025-08-28
2025-08-28