摘要
目的探讨肺耐药蛋白11-反义长链非编码RNA(LncRNA LRP11-AS1)的表达对胃癌(GC)细胞增殖以及免疫逃逸的影响,以及其对微小RNA-149-3p(microRNA-149-3p,miR-149-3p)/叉头状翅膀样转录因子3(FOXP3)的调控机制。方法选取从2019年2月至2022年8月经我院手术切除的80例GC患者的癌组织以及癌旁组织作为研究对象,定量反转录-聚合酶链反应(qRT-PCR)检测GC癌组织和GC癌细胞中LncRNA LRP11-AS1、miR-149-3p、FOXP3的表达。双荧光素酶实验检测LncRNA LRP11-AS1、miR-149-3p、FOXP3的关系。细胞实验分为对照组(Control,正常培养)、NC组(细胞转染ShRNA-NC、antagomir-NC、siRNA-NC)、ShRNA组(细胞转染ShRNA)、ShRNA+antagomir组(细胞转染ShRNA、antagomir)、ShRNA+antagomir+siRNA组(细胞转染ShRNA、antagomir、siRNA)。EDU标记实验检测各组细胞的增殖,AO/EB染色检测各组细胞的凋亡;Transwell小室检测各组细胞的侵袭;Western blot检测各组细胞中FOXP3、波形蛋白(Vimentin)、E-上皮钙粘附素(E-cadherin)的表达。结果LncRNA LRP11-AS1、FOXP3在GC癌组织和GC癌细胞系中的表达明显升高,miR-149-3p的表达明显降低。双荧光素酶实验证实LncRNA LRP11-AS1靶向调控miR-149-3p的表达;miR-149-3p靶向调控FOXP3的表达。抑制LncRNA LRP11-AS1中的表达能明显抑制胃癌细胞的增殖、侵袭,降低细胞中FOXP3和Vimentin、PCNA、TNF-α和INF-γ的表达,上调细胞中E-cadherin和Caspase-3的表达,上调细胞的凋亡率;antagomir能部分解除ShRNA对AGS细胞的抑制作用,而siRNA能部分恢复ShRNA对AGS细胞的抑制作用,差异均具有统计意义(均P<0.05)。结论在GC中,LncRNA LRP11-AS1、FOXP3的表达明显升高,miR-149-3p的表达明显降低,LncRNA LRP11-AS1靶向miR-149-3p调控FOXP3的表达影响AGS细胞的增殖以及免疫逃逸能力。
Objective To investigate the effect of the expression of lung resistance protein 11 antisense long non coding RNA(LncRNA LRP11-AS1)on the proliferation and immune escape of gastric cancer(GC)cells,as well as its regulatory mechanism on microRNA-149-3p(miR-149-3p)/forked/winged helix transcription factor 3(FOXP3).Methods The cancer tissues and paracancerous tissues of 80 patients with GC who were surgically resected in our hospital from February 2019 to August 2022 were selected as subjects.Quantitative reverse transcription-polymerase chain reaction(quantitative reverse transcription-polymerase chain reaction,qRT-PCR)was used to detect the expression of LncRNA LRP11-AS1,miR-149-3p and FOXP3 in GC cancer tissues and GC cancer cells.Double luciferase test was used to detect the relationship among LncRNA LRP11-AS1,miR-149-3p and FOXP3.The cell experiment was divided into control group(Control,normal culture),NC group(cell transfection ShRNA-NC,antagomir-NC,siRNA-NC),ShRNA group(cell transfection ShRNA),ShRNA+antagomir group(cell transfection ShRNA,antagomir),ShRNA+antagomir+siRNA group(cell transfection ShRNA,antagomir,siRNA).EDU labeling test was used to detect the proliferation of cells,AO/EB staining to detect the apoptosis.Transwell chamber was used to detect the invasion of cells.Western blot was used to detect the expression of FOXP3,Vimentin and E-cadherin in cells.Results The expression of LncRNA LRP11-AS1 and FOXP3 in GC cancer tissue and GC cancer cell line was significantly increased,while the expression of miR-149-3p was significantly decreased.Double luciferase assay confirmed that LncRNA LRP11-AS1 targeted the expression of miR-149-3p and miR-149-3p targeted the expression of FOXP3.Inhibition of the expression of LncRNA LRP11-AS1 could significantly inhibit the proliferation and invasion of gastric cancer cells,reduce the expression of FOXP3 and Vimentin,PCNA,TNF-αand INF-γin cells,up-regulate the expression of E-cadherin and Caspase-3 in cells,and up-regulate the apoptosis rate of cells.While,antagomir could partially relieve the inhibitory effect of ShRNA on AGS cells,and siRNA could partially restore the inhibitory effect of ShRNA on AGS cells,the differences were statistically significantly(all P<0.05).Conclusion In GC,the expression of LncRNA LRP11-AS1 and FOXP3 is significantly increased,while the expression of miR-149-3p is significantly decreased.LncRNA LRP11-AS1 targeting miR-149-3p to regulate the expression of FOXP3 affects the proliferation and immune escape ability of AGS cells.
作者
柴立新
徐鑫
潘志坚
CHAI Lixin;XU Xin;PAN Zhijian(Department of Gastrointestinal and Hepatobiliary Surgery,the Affiliated Hospital of Hangzhou Normal University,310000,China)
出处
《现代消化及介入诊疗》
2025年第5期574-582,共9页
Modern Interventional Diagnosis and Treatment in Gastroenterology
