结直肠癌发病相关的lncRNA筛选及GAPLINC对HCT116细胞的作用

来源期刊：广州医药 | 1-6 发布时间：2021-12-21 收稿时间：2025/11/13 17:37:03 阅读量：18

作者：: 罗育其,

温敏杰,

于海涛,

关键词：: lncRNA 结直肠癌细胞迁移及侵袭细胞凋亡; lncRNA Colorectal cancer Invasion and migration of cells Apoptosis of cells

DOI：: 10.3969/j.issn.1000-8535.2019.01.001

收稿时间：: 2018-09-25
修订日期：
接收日期：
引用总数：: 0

目的筛选结直肠癌（CRC）差异性表达的长链非编码RNA（lncRNA）,并进行临床标本验证,研究其对结肠癌细胞HCT116功能的作用。方法利用lncRNA PCR芯片对3对CRC组织和癌旁对照组织筛选差异性表达的lncRNA,确定候选研究lncRNA GAPLINC,RT-qPCR对21例临床样本进行验证其表达的差异性;同时构建GAPLINC表达质粒及其沉默体siRNA转染HCT116细胞,研究其对细胞凋亡、迁移及侵袭能力的影响。结果 lncRNA芯片实验结果提示CRC组织中存在大量的差异性表达的lncRNA,其中GAPLINC在CRC组织表达稳定增加,21例临床样本进一步验证了其在肿瘤组织中表达增加（P<0.05）;转染GAPLINC表达质粒后,HCT116细胞凋亡被抑制,同时其迁移及侵袭能力增强,转染siRNA抑制GAPLINC的表达后,则出现相反的结果。结论利用lncRNA芯片可对CRC差异性表达lncRNA进行批量筛选,GAPLINC在CRC组织中表达稳定增加,具有促癌作用,在CRC发生发展中可能起着重要作用。

Objective To screen the differentiational expression of lncRNA in CRC tissue,confirm it in large simple of clinical specimens,and study its effects on human colorectal cells HCT116 cell line. Methods We screened the lncRNA which expressed differently in 3 CRC tissues and their pair-non carcinour tissues by lncRNA arrays;chose the over expressed lncRNA which played the potential role of oncogene for further researching,and tested the difference in 21 clinical specimens by RT-qPCR. We cultured the HCT166 cells,and then constructed expressed plasmids pcCDNA3.1-GAPLINC and synthesized GAPLINC siRNA,transfected the plasmids and siRNA into HCT116 cells;to study the changes of HCT116 cells behavior,the transwell assays were carried on;the changes of apoptosis of HCT116 cells were tested by flow cytometry. Results There existed many lncRNA which expressed differently between CRC tissues and normal control tissues by lncRNA arrays,there were 21 lncRNA down expressed,and 3 lncRNA up expressed;among these lncRNA,GAPLINC over expressed stably,and its high level of expression was approved in 21 clinical specimens by the test of RT-qPCR. We constructed the expressed plasmids pcCDNA3.1-GAPLINC and synthesizing GAPLINC siRNA successfully;after transfecting pcCDNA3.1-GAPLINC into HCT116 cells,the over expression of GAPLINC increased the migration and invasion of the HCT166 cells （P<0.05）,decreased the proportion of cell apoptosis （P <0.05）;by contraries,knocked down the expression of GAPLINC inhibited invasion and migration of HCT116 cells （P<0.05）,and promoted the apoptosis of the HCT116 cells （P <0.05）. Conclusion It could screen the different expression of lncRNA in large quantities by lncRNA arrays,and GAPLINC expressed highly and stably in CRC tissues. GAPLINC played a role of oncogene,which promoted the proliferation and invasion of CRC cells,and inhibited the apoptosis of CRC cells,which meant playing an important role in the carcinoma and development of CRC.

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