Liu Jie, Shi Zhitao, Ma Yunyun, Fu Liang, Yi Man
Department of Proctology, Affiliated Traditional Chinese Medicine Hospital, Xinjiang Medical University, Urumqi, People's Republic of China.
Department of General Surgery, Affiliated Traditional Chinese Medicine Hospital, Xinjiang Medical University, Urumqi, People's Republic of China.
Onco Targets Ther. 2020 Sep 3;13:8803-8811. doi: 10.2147/OTT.S253470. eCollection 2020.
We aimed at studying the mechanism of MOB1 inhibiting the proliferation and metastasis of colorectal cancer (CRC), to provide a new guidance for the early diagnosis and treatment of CRC.
MOB1 expression level in 68 pairs of CRC tissues and adjacent ones was detected by quantitative real-time polymerase chain reaction (qRT-PCR) analysis, and the associations between the expression level of MOB1 and the clinicopathological indicators as well as the prognosis of CRC patients were analyzed. After constructing CRC cell lines that stably overexpressing or silencing MOB1, the changes of cell proliferation and metastasis ability were examined by Cell Counting Kit (CCK-8) and Transwell assay. In addition, the interaction between MOB1 and PAK2 and how the these two genes affect the biological functions of CRC cell lines were investigated by luciferase assay, qRT-PCR and Western Blot experiments.
Our data showed that MOB1 expression level in CRC tissues was remarkably lower than that in adjacent ones. In comparison to patients of the group of high MOB1 expression, these patients of low MOB1 expression group showed higher incidence of distant or lymph node metastasis and lower survival rate. Cell functional experiments revealed that overexpression of MOB1 markedly attenuated the proliferation and migration ability of CRC cell lines compared to the NC group; In contrast, knockdown of MOB1 enhanced the above-mentioned cell abilities compared to anti-NC group. Luciferase assay verified an interaction between MOB1 and PAK2; and Western blot analysis showed a negative correlation between the expression of the MOB1 and PAK2 protein levels in CRC tissues. Subsequently, we demonstrated that MOB1 interacted with PAK2 to regulate its expression and affected the proliferation and migration capacity of CRC cell lines in vitro.
In summary, the lowly expressed MOB1 in CRC tissues and cell lines may accelerate the proliferation and migration through modulating PAK2 expression.
本研究旨在探讨MOB1抑制结直肠癌（CRC）增殖和转移的机制，为CRC的早期诊断和治疗提供新的指导。
采用定量实时聚合酶链反应（qRT-PCR）分析检测68对CRC组织及其癌旁组织中MOB1的表达水平，并分析MOB1表达水平与CRC患者临床病理指标及预后的相关性。构建稳定过表达或沉默MOB1的CRC细胞系后，通过细胞计数试剂盒（CCK-8）和Transwell实验检测细胞增殖和转移能力的变化。此外，通过荧光素酶报告基因实验、qRT-PCR和蛋白质免疫印迹实验研究MOB1与PAK2之间的相互作用以及这两个基因如何影响CRC细胞系的生物学功能。
我们的数据显示，CRC组织中MOB1的表达水平明显低于癌旁组织。与MOB1高表达组患者相比，MOB1低表达组患者远处或淋巴结转移发生率更高，生存率更低。细胞功能实验表明，与NC组相比，MOB1过表达显著减弱了CRC细胞系的增殖和迁移能力；相反，与抗NC组相比，MOB1基因敲低增强了上述细胞能力。荧光素酶报告基因实验证实了MOB1与PAK2之间存在相互作用；蛋白质免疫印迹分析显示，CRC组织中MOB1和PAK2蛋白水平的表达呈负相关。随后，我们证明MOB1与PAK2相互作用以调节其表达，并在体外影响CRC细胞系的增殖和迁移能力。
总之，CRC组织和细胞系中低表达的MOB1可能通过调节PAK2的表达来加速增殖和迁移。
