[DTX2过表达通过Notch2/Akt轴促进结肠癌细胞的迁移和侵袭]
[DTX2 overexpression promotes migration and invasion of colorectal cancer cells through the Notch2/Akt axis].
作者信息
Ma Z, Zhao X, Zhang X, Xu G, Liu F
机构信息
Department of Anorectal Surgery, Dalian University Affiliated Xinhua Hospital, Dalian 116021, China.
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2023 Mar 20;43(3):340-348. doi: 10.12122/j.issn.1673-4254.2023.03.02.
OBJECTIVE
To investigate the effect of changes in DTX2 expression level on migration and invasion of colorectal cancer (CRC) cells and explore the mechanism.
METHODS
Two CRC cell lines SW620 and LoVo were transfected with a specific shRNA targeting DTX2 (DTX2-shRNA) or a DTX2-overexpressing plasmid (pcDNA-DTX2), and the transfection efficiency was evaluated with RT-qPCR and Western blotting. Scratch and Transwell assays were used to assess the changes in migration and invasion ability of the transfected cells, and the cellular expression levels of Notch2, NICD, AKT, p-Akt and MMP-2/9 proteins were detected with Western blotting. The CRC cells were co-transfected with pcDNA-DTX2 and Notch2 siRNA to assess the effect of Notch2 knockdown on DTX2 overexpression-induced enhancement of cell migration and invasion.
RESULTS
The expression levels of DTX2 at both the mRNA and protein levels were significantly decreased in CRC cells transfected with DTX2- shRNA ( < 0.01) and increased in cells transfected with pcDNA-DTX2 ( < 0.01). Scratch and Transwell assays showed that the migration and invasion abilities of CRC cells were significantly lowered following DTX2 knockdown ( < 0.01) and were enhanced in cells with DTX2 overexpression ( < 0.01). The expression levels of Notch2, NICD, p-Akt and MMP-2 proteins decreased significantly in CRC cells with DTX2 knockdown ( < 0.05) and increased obviously in DTX2-overexpressing cells ( < 0.05). In both of the two CRC cell lines, transfection with Notch2 siRNA obviously reversed the effect of DTX2 overexpression in promoting cell migration and invasion ( < 0.01) and expressions of the related proteins.
CONCLUSION
DTX2 overexpression promotes migration and invasion of CRC cells through the Notch2/Akt axis, suggesting the potential of DTX2 as a new biological indicator of CRC.
目的
研究DTX2表达水平变化对结直肠癌（CRC）细胞迁移和侵袭的影响并探讨其机制。
方法
用靶向DTX2的特异性短发夹RNA（DTX2-shRNA）或DTX2过表达质粒（pcDNA-DTX2）转染两种CRC细胞系SW620和LoVo，通过RT-qPCR和蛋白质免疫印迹法评估转染效率。采用划痕实验和Transwell实验评估转染细胞迁移和侵袭能力的变化，并用蛋白质免疫印迹法检测Notch2、NICD、AKT、p-Akt和MMP-2/9蛋白的细胞表达水平。将CRC细胞与pcDNA-DTX2和Notch2 siRNA共转染，以评估Notch2敲低对DTX2过表达诱导的细胞迁移和侵袭增强的影响。
结果
用DTX2-shRNA转染的CRC细胞中，DTX2的mRNA和蛋白质水平均显著降低（<0.01），而用pcDNA-DTX2转染的细胞中DTX2水平升高（<0.01）。划痕实验和Transwell实验表明，DTX2敲低后CRC细胞的迁移和侵袭能力显著降低（<0.01），而DTX2过表达的细胞中迁移和侵袭能力增强（<0.01）。DTX2敲低的CRC细胞中Notch2、NICD、p-Akt和MMP-2蛋白的表达水平显著降低（<（0.05），在DTX2过表达的细胞中明显升高（<0.05）。在两种CRC细胞系中，用Notch2 siRNA转染均明显逆转了DTX2过表达对细胞迁移和侵袭的促进作用（<0.01）以及相关蛋白的表达。
结论
DTX2过表达通过Notch2/Akt轴促进CRC细胞的迁移和侵袭，提示DTX2作为CRC新的生物学指标的潜力。
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