PDF(Pubmed)
Abstract：
BACKGROUND: Invasion and migration are the irreversible stages of colorectal cancer (CRC). The key is to find a sensitive, reliable molecular marker that can predict the migration of CRC at an early stage. N-myc downstream regulated gene 1 (NDRG1) is a multifunctional gene that has been tentatively reported to have a strong relationship with tumor invasion and migration, however the current molecular role of NDRG1 in CRC remains unknown.
OBJECTIVE: To explore the role of NDRG1 in the development of CRC.
METHODS: NDRG1 stably over-expressed Caco2 cell line was established by lentiviral infection and NDRG1 knock-out Caco2 cell line was established by CRISPR/Cas9. Furthermore, the mRNA and protein levels of NDRG1 in Caco2 cells after NDRG1 over-expression and knockout were detected by real-time polymerase chain reaction and western blot. The cell proliferation rate was measured by the cell counting kit-8 method; cell cycle and apoptosis were detected by flow cytometry; invasion and migration ability were detected by the 24-transwell method.
RESULTS: NDRG1 over-expression inhibited Caco2 proliferation and the cell cycle could be arrested at the G1/S phase when NDRG1 was over-expressed, while the number of cells in the G2 phase was significantly increased when NDRG1 was knocked out. This suggests that NDRG1 inhibited the proliferation of Caco2 cells by arresting the cell cycle in the G1/S phase. Our data also demonstrated that NDRG1 promotes early cell apoptosis. Invasion and migration of cells were extensively inhibited when NDRG1 was over-expressed.
CONCLUSIONS: NDRG1 inhibits tumor progression in Caco2 cells which may represent a potential novel therapeutic strategy for the treatment of CRC.
OBJECTIVE: To explore the role of NDRG1 in the development of CRC.
METHODS: NDRG1 stably over-expressed Caco2 cell line was established by lentiviral infection and NDRG1 knock-out Caco2 cell line was established by CRISPR/Cas9. Furthermore, the mRNA and protein levels of NDRG1 in Caco2 cells after NDRG1 over-expression and knockout were detected by real-time polymerase chain reaction and western blot. The cell proliferation rate was measured by the cell counting kit-8 method; cell cycle and apoptosis were detected by flow cytometry; invasion and migration ability were detected by the 24-transwell method.
RESULTS: NDRG1 over-expression inhibited Caco2 proliferation and the cell cycle could be arrested at the G1/S phase when NDRG1 was over-expressed, while the number of cells in the G2 phase was significantly increased when NDRG1 was knocked out. This suggests that NDRG1 inhibited the proliferation of Caco2 cells by arresting the cell cycle in the G1/S phase. Our data also demonstrated that NDRG1 promotes early cell apoptosis. Invasion and migration of cells were extensively inhibited when NDRG1 was over-expressed.
CONCLUSIONS: NDRG1 inhibits tumor progression in Caco2 cells which may represent a potential novel therapeutic strategy for the treatment of CRC.
摘要：
背景：侵袭和迁移是结直肠癌（CRC）的不可逆阶段。关键是要找到一个敏感的，可靠的分子标记，可以在早期预测CRC的迁移。N-myc下游调控基因1(NDRG1)是一种多功能基因,目前已初步报道与肿瘤的侵袭和迁移有很强的关系,然而,目前NDRG1在CRC中的分子作用尚不清楚.
目的：探讨NDRG1在CRC发生发展中的作用。
方法：通过慢病毒感染建立NDRG1稳定过表达的Caco2细胞系，并通过CRISPR/Cas9建立NDRG1敲除的Caco2细胞系。此外，通过实时聚合酶链反应和Westernblot检测NDRG1过表达和敲除后Caco2细胞中NDRG1的mRNA和蛋白水平。细胞计数试剂盒-8法测定细胞增殖率；流式细胞术检测细胞周期和凋亡；24-transwell法检测侵袭和迁移能力。
结果：NDRG1过表达抑制了Caco2的增殖，NDRG1过表达时，细胞周期可以阻滞在G1/S期，而当敲除NDRG1时，G2期的细胞数量显着增加。这表明NDRG1通过将细胞周期阻滞在G1/S期来抑制Caco2细胞的增殖。我们的数据还表明NDRG1促进早期细胞凋亡。当NDRG1过表达时，细胞的侵袭和迁移被广泛抑制。
结论：NDRG1抑制Caco2细胞中的肿瘤进展，这可能是治疗CRC的潜在新治疗策略。
目的：探讨NDRG1在CRC发生发展中的作用。
方法：通过慢病毒感染建立NDRG1稳定过表达的Caco2细胞系，并通过CRISPR/Cas9建立NDRG1敲除的Caco2细胞系。此外，通过实时聚合酶链反应和Westernblot检测NDRG1过表达和敲除后Caco2细胞中NDRG1的mRNA和蛋白水平。细胞计数试剂盒-8法测定细胞增殖率；流式细胞术检测细胞周期和凋亡；24-transwell法检测侵袭和迁移能力。
结果：NDRG1过表达抑制了Caco2的增殖，NDRG1过表达时，细胞周期可以阻滞在G1/S期，而当敲除NDRG1时，G2期的细胞数量显着增加。这表明NDRG1通过将细胞周期阻滞在G1/S期来抑制Caco2细胞的增殖。我们的数据还表明NDRG1促进早期细胞凋亡。当NDRG1过表达时，细胞的侵袭和迁移被广泛抑制。
结论：NDRG1抑制Caco2细胞中的肿瘤进展，这可能是治疗CRC的潜在新治疗策略。
