Wang Zhuo, Hu Tao, Jin Chengwu, Yu Jiangui, Zhu Dongqiang, Liu Jian
Department of General Surgery, Chengdu Fifth People's Hospital, Chengdu, China.
J Gastrointest Oncol. 2020 Oct;11(5):899-910. doi: 10.21037/jgo-20-387.
Colon cancer is one of the major causes of morbidity and mortality worldwide. MicroRNAs (miRNAs) play important functions in the growth and metastasis of colon cancer. This study aimed to investigate the anti-tumor effect of micro ribonucleic acid 539-3p (miR-539-3p) on colon cancer via regulation of cell viability, motility, and nude mouse tumorigenicity with cyclin-dependent kinase 14 (CDK14) inhibition.
The target relationship between miR-539-3p and CDK14 was predicted using TargetScan software, and were detected by luciferase reporter assay. Cell counting kit-8 (CCK-8) assay and flow cytometry were employed to examine cell proliferation and apoptosis. Western blotting was employed to measure the protein expression levels of p27, cleaved caspase-3, and epithelial (E)- and neural (N)-cadherin. The effect of miR-539-3p on tumor growth was evaluated by establishing a xenograft tumor model in nude mice.
The target relationship of CDK14 and miR-539-3p was identified as a negative regulator. Overexpression of miR-539-3p significantly inhibited SW620 and SW480 cell proliferation, promoted cell apoptosis, and suppressed cell invasion by targeting CDK14. The xenograft tumor model showed that the overexpression of miR-539-3p reduced tumor weight and volume. Immunohistochemical staining revealed that the overexpression of miR-539-3p inhibited the expression of Ki67 and E-cadherin. Additionally, terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining showed that overexpression of miR-539-3p induced apoptosis.
Overexpression of miR-539-3p inhibited SW620 and SW480 cell proliferation, promoted cell apoptosis, and suppressed cell invasion by targeting CDK14. Therefore, miR-539-3p may be a useful diagnostic and therapeutic biomarker for colon cancer.
结肠癌是全球发病和死亡的主要原因之一。微小RNA（miRNA）在结肠癌的生长和转移中发挥重要作用。本研究旨在通过抑制细胞周期蛋白依赖性激酶14（CDK14）来调节细胞活力、运动能力和裸鼠致瘤性，从而探讨微小核糖核酸539-3p（miR-539-3p）对结肠癌的抗肿瘤作用。
使用TargetScan软件预测miR-539-3p与CDK14之间的靶标关系，并通过荧光素酶报告基因检测进行验证。采用细胞计数试剂盒-8（CCK-8）检测法和流式细胞术检测细胞增殖和凋亡情况。采用蛋白质免疫印迹法检测p27、裂解的半胱天冬酶-3以及上皮（E）-钙黏蛋白和神经（N）-钙黏蛋白的蛋白表达水平。通过在裸鼠中建立异种移植肿瘤模型来评估miR-539-3p对肿瘤生长的影响。
确定CDK14与miR-539-3p的靶标关系为负调控。miR-539-3p的过表达通过靶向CDK14显著抑制SW620和SW480细胞增殖，促进细胞凋亡，并抑制细胞侵袭。异种移植肿瘤模型显示，miR-539-3p的过表达降低了肿瘤重量和体积。免疫组织化学染色显示，miR-539-3p的过表达抑制了Ki67和E-钙黏蛋白的表达。此外，末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）染色显示，miR-539-3p的过表达诱导了细胞凋亡。
miR-539-3p的过表达通过靶向CDK14抑制SW620和SW480细胞增殖，促进细胞凋亡，并抑制细胞侵袭。因此，miR-539-3p可能是结肠癌有用的诊断和治疗生物标志物。
