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河北医学  2023, Vol. 29 Issue (11): 1777-1783    DOI: 10.3969/j.issn.1006-6233.2023.11.04
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LncRNA MAFG-AS1调节miR-331-3p/SERPINE1轴对胃癌细胞恶性生物学行为的影响
宋玉涛, 王峰, 李凡, 冯浩
河北省定州市人民医院, 河北 定州 073000
Impact of LncRNA MAFG-AS1 on the Malignant Biological Behavior of Gastric Cancer Cells by Regulating the miR-331-3p/SERPINE1 Axis
SONG Yutao, WANG Feng, LI Fan, et al
The People's Hospital of Dingzhou, Hebei Dingzhou 073000, China
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摘要 目的: 探讨长链非编码RNA(LncRNA)重组人V-maf肌腱膜纤维肉瘤癌基因G反义RNA1(MAFG-AS1)调节miR-331-3p/SERPINE1轴对胃癌细胞恶性生物学行为的影响。方法: qRT-PCR检测人胃癌组织(n=30)、癌旁组织(n=30)、胃黏膜正常上皮细胞GES-1、人胃癌细胞BGC-823、SGC-7901、MGC-803中MAFG-AS1、miR-331-3p、SERPINE1表达;将BGC-823细胞随机分为ctrl组、pcDNA3.1组、pcLncRNA MAFG-AS1组、pcLncRNA MAFG-AS1+miR-NC组、pcLncRNA MAFG-AS1+ miR-331-3p组;qRT-PCR检测各组BGC-823细胞MAFG-AS1、miR-331-3p、SERPINE1水平;CCK-8法检测各组BGC-823细胞活力;EDU法检测各组BGC-823细胞增殖;Transwell检测各组BGC-823细胞迁移和侵袭;western blot检测各组BGC-823细胞SERPINE1、PCNA、MMP-2表达;双荧光素酶报告基因实验分别验证MAFG-AS1和miR-331-3p、miR-331-3p和SERPINE1的关系。结果: 与癌旁组织相比,胃癌组织中MAFG-AS1、SERPINE1 mRNA表达显著性升高(3.01±0.37 vs 1.02±0.12,2.49±0.26 vs 1.01±0.10,t=28.022、29.000,P<0.05),miR-331-3p表达显著性降低(0.32±0.03 vs 1.03±0.07,t=51.063,P<0.05)。与GES-1细胞比较,不同胃癌细胞中MAFG-AS1、SERPINE1 mRNA表达显著性升高,miR-331-3p表达显著性降低(P<0.05),且BGC-823细胞变化结果更显著,后续实验选择BGC-823细胞。与ctrl组、pcDNA3.1组比较,pcLncRNA MAFG-AS1组BGC-823细胞MAFG-AS1、SERPINE1 mRNA水平、细胞活力A值、EDU阳性细胞率、迁移与侵袭细胞数、SERPINE1、PCNA、MMP-2蛋白水平均显著性升高,miR-331-3p mRNA水平显著性降低(P<0.05);与pcLncRNA MAFG-AS1组、pcLncRNA MAFG-AS1+miR-NC组比较,pcLncRNA MAFG-AS1+miR-331-3p组BGC-823细胞SERPINE1 mRNA水平、细胞活力A值、EDU阳性细胞率、迁移与侵袭细胞数、SERPINE1、PCNA、MMP-2蛋白水平均显著性降低,miR-331-3p mRNA水平显著性升高(P<0.05);MAFG-AS1靶向负调控miR-331-3p表达,miR-331-3p靶向负调控SERPINE1表达。结论: 过表达LncRNA MAFG-AS1可能通过靶向下调miR-331-3p来上调SERPINE1表达,进而促进BGC-823细胞增殖、迁移和侵袭。
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宋玉涛
王峰
李凡
冯浩
关键词 LncRNA MAFG-AS1miR-331-3pSERPINE1胃癌细胞    
AbstractObjective: To investigate the impact of the long non-coding RNA (LncRNA) recombinant human v-maf musculoaponeurotic fibrosarcoma oncogene homolog G antisense RNA1 (MAFG-AS1) on the malignant biological behavior of gastric cancer cells by regulating the miR-331-3p/SERPINE1 axis. Methods: QRT-PCR was applied to detect the expression of MAFG-AS1,miR-331-3p,and SERPINE1 in gastric cancer tissues (n=30),paracancerous tissues (n=30),human gastric mucosa normal epithelial cells GES-1,human gastric cancer cells BGC-823,SGC-7901,and MGC-803; BGC-823 cells were randomly grouped into ctrl group,pcDNA3.1 group,pcLncRNA MAFG-AS1 group,pcLncRNA MAFG-AS1+miR-NC group,and pcLncRNA MAFG-AS1+miR-331-3p group; qRT-PCR was applied to detect the levels of MAGG-AS1,miR-331-3p,and SERPINE1 in BGC-823 cells in each group; CCK-8 method was applied to detect the activity of BGC-823 cells in each group; EDU method was applied to detect the proliferation of BGC-823 cells in each group; Transwell was applied to detect the migration and invasion of BGC-823 cells in each group; Western blot was applied to detect the expression of SERPINE1,PCNA,and MMP-2 in BGC-823 cells in each group; Double Luciferase reporter gene experiment was applied to verify the relationship between MAFG-AS1 and miR-331-3p,miR-331-3p and SERPINE1. Results: In comparison to adjacent tissues,gastric cancer tissues exhibited significantly increased expressions of MAFG-AS1 and SERPINE1 mRNA (3.01±0.37 vs 1.02±0.12 and 2.49±0.26 vs1.01±0.10; t=28.022 and 29.000; P<0.05) and markedly decreased miR-331-3p expression (0.32±0.03 vs 1.03±0.07; t=51.063; P<0.05).Furthermore,various gastric cancer cells displayed notably higher MAFG-AS1 and SERPINE1 mRNA expressions and lower miR-331-3p expression in comparison to GES-1 cells (P<0.05).Among the gastric cancer cell lines,BGC-823 cells exhibited the most significant changes,so we selected BGC-823 cells for subsequent experiments.Compared to the control group and pcDNA3.1 group,the pcLncRNA MAFG-AS1 group showed significantly higher levels of MAFG-AS1,SERPINE1 mRNA,cell viability (A value),EDU-positive cell rate,numbers of migration and invasion cells,as well as SERPINE1,PCNA,and MMP-2 protein levels,while miR-331-3p mRNA levels were significantly lower (P<0.05).In comparison to the pcLncRNA MAFG-AS1 group and the pcLncRNA MAFG-AS1+miR-NC group,the pcLncRNA MAFG-AS1+miR-331-3p group showed significantly lower SERPINE1 mRNA levels,cell viability (A value),EDU-positive cell rate,numbers of migration and invasion cells,as well as SERPINE1,PCNA,and MMP-2 protein levels,whereas miR-331-3p mRNA levels were significantly higher (P<0.05).MAFG-AS1 negatively regulated miR-331-3p expression,and miR-331-3p negatively regulated SERPINE1 expression. Conclusion: Overexpression of LncRNA MAFG-AS1 may upregulate SERPINE1 expression by negatively regulating miR-331-3p,thereby promoting proliferation,migration and invasion of BGC-823 cells.
Key wordsLncRNA MAFG-AS1    miR-331-3p    SERPINE1    Gastric cancer cells
    
基金资助:河北省医学科学研究课题,(编号:20220437)
通讯作者: 冯浩   
引用本文:   
宋玉涛, 王峰, 李凡, 冯浩. LncRNA MAFG-AS1调节miR-331-3p/SERPINE1轴对胃癌细胞恶性生物学行为的影响[J]. 河北医学, 2023, 29(11): 1777-1783.
SONG Yutao, WANG Feng, LI Fan, et al. Impact of LncRNA MAFG-AS1 on the Malignant Biological Behavior of Gastric Cancer Cells by Regulating the miR-331-3p/SERPINE1 Axis. HeBei Med, 2023, 29(11): 1777-1783.
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