lncRNA-ANRIL影响胃癌细胞侵袭转移的功能和机制研究

Research of lncRN-A-ANRIL affecting gastric cancer invasion and metastasis

目的探讨TET(ten-eleven translocation)2和lncRNA-ANRIL对胃癌细胞侵袭转移的影响。方法采用定量逆转录聚合酶链反应(quantitative reverse transcription polymerase chain reaction,q RT-PCR)检测胃癌组织样本和相应癌旁组织中TET2和lncRNA-ANRIL的表达量;构建TET2和lncRNA-ANRIL的慢病毒载体,并转染胃癌细胞MGC803,于胃癌细胞MGC803中敲减TET2后,采用q RT-PCR检测lncRNA-ANRIL的表达量,以探究二者之间存在的调控关系;采用荧光素酶报告基因检测TET2对lncRNA-ANRIL启动子序列的影响;采用Transwell实验检测TET2和lncRNA-ANRIL对胃癌细胞MGC803侵袭转移能力的影响。结果 TET2在胃癌组织中低表达,而lncRNA-ANRIL在胃癌组织中高表达;在胃癌细胞MGC803中敲减TET2后,lncRNA-ANRIL的表达量显著上调。荧光素酶报告基因检测进一步验证了TET2能够通过抑制lncRNA-ANRIL的启动子活性,调控lncRNA-ANRIL的表达;Transwell实验结果表明,敲减TET2后,胃癌细胞MGC803的侵袭转移能力显著增强,而敲减lncRNA-ANRIL则能明显抑制胃癌细胞MGC803的侵袭转移能力,敲减TET2后再敲减lncRNA-ANRIL能够减弱敲减TET2增强胃癌细胞MGC803的侵袭转移能力的效果。结论 TET2和lncRNA-ANRIL在胃癌细胞的侵袭转移中发挥重要作用,本研究为胃癌靶向治疗提供了理论依据。

Objective To investigate the effects of ten-eleven translocation(TET)2and lncRNA-ANRIL on gastric cancer cells invasion and metastasis.Method The expression of TET2and lncRNA-ANRIL in gastric cancer tissues and corresponding adjacent tissues were detected by quantitative real-time polymerase chain reaction(qRT-PCR).The lentiviral vectors for TET2and lncRNA-ANRIL were constructed and transfected into MGC803cells and then the expression of lncRNA-ANRIL was detected by qRT-PCR in order to explore the relationship between TET2and lncRNA-ANRIL.The luciferase reporter assay was used to detect the expression of TET2in the gastric cancer cell MGC803and indicated that TET2could affect the activity of the promoter of lncRNA-ANRIL.Transwell experiment was used to study the effect of TET2and lncRNA-ANRIL on invasion and metastasis of gastric cancer cell MGC803.Result The expression of TET2was downregulated in gastric cancer tissues,while lncRNA-ANRIL was highly expressed in gastric cancer tissues.The expression of lncRNA-ANRIL was significantly up-regulated in gastric cancer cell MGC803after knockdown of TET2.The luciferase reporter assay showed that TET2could regulate the expression of lncRNA-ANRIL by inhibiting the activity of the activity of the promoter of lncRNA-ANRIL.Transwell experiments showed that the invasion and metastasis ability of gastric cancer cells was significantly enhanced after knockdown of TET2,while knockdown of lncRNA-ANRIL could rescue the enhancement.Conclusion TET2and lncRNAANRIL play an important role in the invasion and metastasis of gastric cancer cells.Our study provides a theoretical basis for gastric cancer targeted therapy.