长链非编码RNA AFAP1-AS1的过表达对胃癌细胞增殖和迁移的影响
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摘要
目的探讨长链非编码RNA AFAP1-AS1的过表达对胃癌细胞增殖和迁移的影响。方法采用定量逆转录聚合酶链式反应(qRT-PCR)检测52例胃癌及癌旁标本(2015年5月～2017年1月在南京医科大学附属南京第一医院收集)和6例胃癌细胞株中lncRNA AFAP1-AS1的表达水平。小干扰RNA(siRNA)用于抑制胃癌细胞系中的AFAP1-AS1表达。结果胃癌组织中的AFAP1-AS1水平显著高于相应的癌旁组织,差异有统计学意义(P<0.01)。胃癌细胞系AGS、SGC7901、MGC803和BGC8231的表达量显著高于正常胃黏膜细胞系GES-1,差异有统计学意义(P<0.05或P<0.01)。AFAP1-AS1的表达水平与胃癌患者临床分期(χ~2=7.137,P=0.008)和淋巴结转移(χ~2=6.656,P=0.010)有关。CCK8和克隆集落形成实验显示,用siRNA敲降AFAP1-AS1的表达后,胃癌细胞系BGC823和MGC803的增殖速率显著降低,差异有统计学意义(P<0.01或P<0.001)。细胞划痕实验显示,用siRNA敲降AFAP1-AS1的表达后,胃癌细胞系BGC823和MGC803的增殖速率显著降低,差异有统计学意义(P<0.05)。结论 AFAP1-AS1是一种参与胃癌进展的新型生物标志物,可能为治疗性干预提供潜在的预后生物标志物和潜在靶点。
Objective To explore the influence of overexpression of LncRNA AFAP1-AS1 on proliferation and migration of gastric cancer cells. Methods The expression levels of lncRNA AFAP1-AS1 in 52 cases of gastric cancer and paracancerous tissues which were collected from May 2015 to January 2017 and 6 gastric cancer cell lines were evaluated by quantitative reverse transcriptase polymerase chain reaction(q RT-PCR). Small interfering RNA(siRNA) was used to suppress AFAP1-AS1 expression in gastric cancer cell lines. Results The level of AFAP1-AS1 in gastric cancer tissues was significantly higher than that in adjacent adjacent tissues, with significant difference(P<0.01). The expression levels of AGS, SGC7901, MGC803 and BGC8231 in gastric cancer cell lines were significantly higher than those of normal gastric mucosal cell line GES-1, the difference was statistically significant(P<0.05 or P<0.01). The expression level of AFAP1-AS1 was related to clinical stage(χ~2=7.137, P=0.008) and lymph node metastasis(χ~2=6.656 P=0.010).CCK8 and colony colony formation experiments confirmed that the proliferation rate of gastric cancer cell lines BGC823 and MGC803 significantly decreased after knockdown of AFAP1-AS1 expression which induced by siRNA, the differences were statistically significant(P<0.01 or P<0.001). Cell scratch assay confirmed that the proliferation rate of gastric cancer cell lines BGC823 and MGC803 significantly decreased after siRNA knockdown of AFAP1-AS1 expression, the difference was statistically significant(P<0.05). Conclusion AFAP1-AS1 is a novel biomarker involved in gastric cancer progression, which may provide a potential prognostic biomarker and a prospective target for therapeutic intervention.
Objective To explore the influence of overexpression of LncRNA AFAP1-AS1 on proliferation and migration of gastric cancer cells. Methods The expression levels of lncRNA AFAP1-AS1 in 52 cases of gastric cancer and paracancerous tissues which were collected from May 2015 to January 2017 and 6 gastric cancer cell lines were evaluated by quantitative reverse transcriptase polymerase chain reaction(q RT-PCR). Small interfering RNA(siRNA) was used to suppress AFAP1-AS1 expression in gastric cancer cell lines. Results The level of AFAP1-AS1 in gastric cancer tissues was significantly higher than that in adjacent adjacent tissues, with significant difference(P<0.01). The expression levels of AGS, SGC7901, MGC803 and BGC8231 in gastric cancer cell lines were significantly higher than those of normal gastric mucosal cell line GES-1, the difference was statistically significant(P<0.05 or P<0.01). The expression level of AFAP1-AS1 was related to clinical stage(χ~2=7.137, P=0.008) and lymph node metastasis(χ~2=6.656 P=0.010).CCK8 and colony colony formation experiments confirmed that the proliferation rate of gastric cancer cell lines BGC823 and MGC803 significantly decreased after knockdown of AFAP1-AS1 expression which induced by siRNA, the differences were statistically significant(P<0.01 or P<0.001). Cell scratch assay confirmed that the proliferation rate of gastric cancer cell lines BGC823 and MGC803 significantly decreased after siRNA knockdown of AFAP1-AS1 expression, the difference was statistically significant(P<0.05). Conclusion AFAP1-AS1 is a novel biomarker involved in gastric cancer progression, which may provide a potential prognostic biomarker and a prospective target for therapeutic intervention.
引文
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[4]Yoon SH,Kim YH,Lee YJ,et al.Tumor heterogeneity in human epidermal growth factor receptor 2(HER2)-positive advanced gastric cancer assessed by CT texture analysis:association with survival after trastuzumab treatment[J].PLoS One,2016,11(8):e0161278.
[5]吴钢,赵昌杰,姜琦,等.胃癌血清标志物的临床意义及研究进展[J].中国当代医药,2016,23(14):10-12.
[6]Esteller M.Non-coding Rnas in human disease[J].Nat Rev Genet,2011,12(12):861-874.
[7]Gupta RA,Shah N,Wang KC,et al.Long non-coding RNA HOTAIR reprograms chromatin state to promote cancer metastasis[J].Nature,2010,464(7291):1071-1076.
[8]Li W,Zheng J,Deng J,et al.Increased levels of the long intergenic non-protein coding RNA POU3F3 promote DNA methylation in esophageal squamous cell carcinoma cells[J].Gastroenterology,2014,146(7):1714-1726.
[9]崔大炜,姜峰,陈大欣.LncRNA MEG3在甲状腺乳头状癌中的表达及对增殖、侵袭与转移能力的影响[J].中国医药导报,2017,14(32):15-18,42.
[10]Yang SL,Lin RX,Si LH,et al.Expression and functional role of long non-coding RNA AFAP1-AS1 in ovarian cancer[J].Eur Rev Med Pharmacol Sci,2016,20(24):5107-5112.
[11]Ma F,Wang SH,Cai Q,et al.Overexpression of LncRNA AFAP1-AS1 predicts poor prognosis and promotes cells proliferation and invasion in gallbladder cancer[J].Biomed Pharmacother,2016,(84):1249-1255.
[12]Luo HL,Huang MD,Guo JN,et al.AFAP1-AS1 is upregulated and promotes esophageal squamous cell carcinoma cell proliferation and inhibits cell apoptosis[J].Cancer Med,2016,5(10):2879-2885.
[13]Han X,Wang L,Ning Y,et al.Long non-coding RNA AFAP1-AS1 facilitates tumor growth and promotes metastasis in colorectal cancer[J].Biol Res,2016,49(1):36.
[14]Fu XL,Liu DJ,Yan TT,et al.Analysis of long non-coding RNA expression profiles in pancreatic ductal adenocarcinoma[J].Sci Rep,2016,(6):33 535.
[15]Mercer TR,Dinger ME,Mattick JS.Long non-coding RNAs:insights into functions[J].Nat Rev Genet,2009,10(3):155-159.
[16]赵志国,张力平.miRNA和lncRNA相互作用与恶性肿瘤相关性的研究进展[J].现代肿瘤医学,2018,26(18):2991-2994.
[17]Deng J,Liang Y,Liu C,et al.The up-regulation of long noncoding RNA AFAP1-AS1 is associated with the poor prognosis of NSCLC patients[J].Biomed Pharmacother,2015,75:8-11.
[18]Bo H,Gong Z,Zhang W,et al.Upregulated long non-coding RNA AFAP1-AS1 expression is associated with progression and poor prognosis of nasopharyngeal carcinoma[J].Oncotarget,2015,6(24):20 404-20 418.
[19]Lu X,Zhou C,Li R,et al.Critical role for the long noncoding RNA AFAP1-AS1 in the proliferation and metastasis of hepatocellular carcinoma[J].Tumor Bio,2016,37(7):1-9.
[20]Huang JY,Xu YY,L i M,et al.The prognostic impact of occult lymph node metastasis in node-negative gastric cancer:a systematic review and meta-analysis[J].Ann Surg Oncol,2013,20(12):3927-3934.
[21]Li J,Yi CH,Hu YT,et al.TNM staging of colorectal cancer should be reconsidered according to weighting of the T stage:verification based on a 25-year follow-up[J].Medicine,2016,95(6):e2711.