微小RNA-4486通过靶向丝裂原活化蛋白激酶激酶4抑制p38丝裂原活化蛋白激酶/应激活化蛋白激酶信号通路的激活，从而抑制结直肠癌增殖。

miR-4486 inhibits colorectal cancer proliferation via targeting MAP2K4 to inhibit the activation of the p38MAPK/JNK signaling.

作者信息

Wang Weiwei, Chen Linxia, Xu Feipeng, Chen Rihong, Li Qidong, Zou Lirui, Hu Honghui, Zhu Wenjing

机构信息

Department of Gastrointestinal Surgery, the Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, 524001, China.

Department of Operating Room, the Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong, 524001, China.

出版信息

Heliyon. 2024 Oct 3;10(21):e38926. doi: 10.1016/j.heliyon.2024.e38926. eCollection 2024 Nov 15.

OBJECTIVE

Since MAP2K4 was reportedly involved in colorectal cancer development and the p38MAPK/JNK signaling transcription, this study aimed to investigate the mechanism by which the microRNA (miR)-4486 acts on colorectal cell proliferation.

METHODS

RT-PCR was conducted to measure the expression levels of the MAP2K4 and miR-4486 in NCM460, SW1116, and HCT116 cells. TargetScanHuman site anticipated that MAP2K4 may be a target of miR-4486. The dual-luciferase reporter assay confirmed their relationship. After plasmids of miR-4486 mimic and si-MAP2K4 transfection, MAP2K4 was quantified again, The CCK-8 assay was carried out to assess cell proliferation, while Scratch and Transwell assays were used to evaluate cell migration and invasion. Finally, miR-4486 mimic and SB203580 were applied in HCT116 and SW1116 cells separately or in combination. CCK-8, Scratch and Transwell assay were performed again. In addition, the proteins including c-capase3, Bax, Bcl2, MAP2K4, and the p38MAPK/JNK signaling-related proteins expression levels were quantified by Western blot (WB).

RESULTS

Compared with the NCM460 cells, the expression level of MAP2K4 was elevated, while the expression level of miR-4486 was reduced in SW1116 and HCT116 cells. The results showed that elevated levels of miR-4486 suppressed cell proliferation, migration, and invasion in colorectal cells by downregulating MAP2K4 expression. miR-4486 mimic showed similar effects to SB203580, which promoted colorectal cell apoptosis and inhibited the p38 MAPK/JNK signaling transcription.

CONCLUSION

miR-4486 may target MAP2K4 to inhibit colorectal cell proliferation by inhibiting the activation of the p38/JNK signaling pathway.

目的

鉴于据报道MAP2K4参与结直肠癌的发展以及p38丝裂原活化蛋白激酶/应激活化蛋白激酶（p38MAPK/JNK）信号转导，本研究旨在探究微小RNA（miR）-4486作用于结肠直肠细胞增殖的机制。

方法

采用逆转录聚合酶链反应（RT-PCR）检测正常结肠黏膜上皮细胞系（NCM460）、人结肠癌细胞系（SW1116）和人结肠癌细胞系（HCT116）中MAP2K4和miR-4486的表达水平。TargetScanHuman网站预测MAP2K4可能是miR-4486的一个靶点。双荧光素酶报告基因检测证实了它们之间的关系。在转染miR-4486模拟物和小干扰RNA（si）-MAP2K4质粒后，再次对MAP2K4进行定量分析，采用细胞计数试剂盒-8（CCK-8）检测评估细胞增殖，采用划痕实验和Transwell实验评估细胞迁移和侵袭。最后，将miR-4486模拟物和SB203580分别或联合应用于HCT116和SW1116细胞。再次进行CCK-8、划痕实验和Transwell实验。此外，通过蛋白质免疫印迹法（WB）对包括半胱天冬酶3（c-capase3）、凋亡相关蛋白（Bax）、抗凋亡蛋白（Bcl2）、MAP2K4以及p38MAPK/JNK信号相关蛋白的表达水平进行定量分析。

结果

与NCM460细胞相比，SW1116和HCT116细胞中MAP2K4的表达水平升高，而miR-4486的表达水平降低。结果表明，miR-4486水平升高可通过下调MAP2K4表达抑制结肠直肠细胞的增殖、迁移和侵袭。miR-4486模拟物显示出与SB203580相似的作用，即促进结肠直肠细胞凋亡并抑制p38丝裂原活化蛋白激酶/应激活化蛋白激酶（p38 MAPK/JNK）信号转导。