lncRNA TTN-AS1通过靶向miR-204-3p调控胃癌细胞迁移、侵袭及上皮-间质转化

目的：探讨长链非编码RNA肌联蛋白反义RNA 1（lncRNA TTN-AS1）是否可通过靶向微小RNA-204-3p（miR-204-3p）调控胃癌细胞迁移、侵袭及上皮-间质转化（EMT）。方法：采用qRT-PCR法检测胃癌组织、癌旁组织中TTN-AS1、miR-204-3p的表达量；体外培养人胃癌细胞AGS，分别将si-NC、si-TTN-AS1、miR-NC、miR-204-3p mimics、si-TTN-AS1与anti-miR-NC、si-TTN-AS1与anti-miR-204-3p转染至AGS细胞；采用qRTPCR法检测AGS细胞中TTN-AS1、miR-204-3p的表达量；采用Transwell小室实验检测迁移及侵袭能力；双荧光素酶报告实验检测TTN-AS1、miR-204-3p的靶向关系。结果：胃癌组织中TTN-AS1的表达水平比癌旁组织增加约2.90倍（P ＜0.05），miR-204-3p的表达水平比癌旁组织减少约0.57倍（P ＜0.05）；转染si-TTN-AS1或转染miR-204-3p mimics可明显减少迁移及侵袭细胞数（P ＜0.05）；双荧光素酶报告实验证实TTN-AS1可靶向结合miR-204-3p；共转染si-TTN-AS1与anti-miR-204-3p可明显增加迁移及侵袭细胞数（P ＜0.05）。结论：抑制TTN-AS1表达可通过上调miR-204-3p的表达从而抑制胃癌细胞迁移、侵袭及EMT。

The mechanism of lncRNA TTN-AS1 in regulating gastric cancer cell migration,invasion and EMT by targeting miR-204-3p

Objective: To explore the role of lncRNA TTN-AS1 in regulation the migration, invasion and epithelial-mesenchymal transition (EMT) of gastric cancer cells by targeting miR-204-3p. Methods: qRT-PCR method was used to detect the expression of TTN-AS1 and miR-204-3p in gastric cancer tissues and adjacent tissues. In vitro culture of human gastric cancer cell AGS, si-NC, si-TTN-AS1, miR-NC, miR-204-3p mimics, si-TTN-AS1 and anti-miR-NC, si-TTN-AS1 and anti- miR-204-3p were transfected into AGS cells. The qRT-PCR method was used to detect the expression of TTN-AS1 and miR-204-3p in AGS cells. The Transwell chamber assay was employed to monitor migration and invasion capabilities. The dual luciferase reporter experiment was used to detect the targeting relationship of TTN-AS1 and miR-204-3p. Results: The expression level of TTNAS1 in gastric cancer tissues was increased by about 2.90 times (P<0.05), and the expression level of miR-204-3p was decreased by about 0.57 times compared with adjacent tissues (P<0.05). Transfection of si-TTN-AS1 or transfection of miR-204-3p mimics could significantly reduce the number of migrating and invading cells (P<0.05). TTN-AS1 could target miR-204-3p, which was confirmed by the dual luciferase report experiment.Co-transfection of si-TTN-AS1 and anti-miR-204-3p could significantly increase the number of migrating and invading cells (P<0.05). Conclusion: The inhibition of TTN-AS1 expression could inhibit gastric cancer cell migration, invasion and EMT transformation by up-regulating the expression of miR-204-3p.