MicroRNA-323a-3p Negatively Regulates NEK6 in Colon Adenocarcinoma Cells.

Zhongshi Hong,Zhichuan Chen,Zesheng Shi,Jianpeng Pan,Chengzhi Qiu,Chunxiao Wang,Zhiqian Zhang

doi:10.1155/2022/7007718

Abstract

Objective The activity of NEK6 is enhanced in several cancer cells, including colon adenocarcinoma (COAD) cells. However, there are few reports on the microRNA (miRNA/miR) regulation of NEK6. In this study, we aimed to investigate the effects of miRNAs targeting NEK6 in COAD cells. Methods Public data and online analysis sites were used to analyze the expression levels of NEK6 and miR-323a-3p in COAD tissues as well as the relationship between NEK6 or miR-323a-3p levels and survival in patients with COAD and to predict miRNAs targeting NEK6. Real-time polymerase chain reaction and western blotting were performed to determine the levels of NEK6 and miR-323a-3p in COAD cells. The targeting of NEK6 by miR-323a-3p was verified using a dual-luciferase reporter assay. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, 5-ethynyl-2′-deoxyuridine assay, propidium iodide (PI) staining, annexin V-fluorescein isothiocyanate/PI staining, and transwell assay were employed to test the proliferation, apoptosis, migration ability, and invasiveness of COAD cells. Results In COAD cells, NEK6 was highly expressed, whereas miR-323a-3p was expressed at low levels and negatively regulated NEK6. Upregulating the level of miR-323a-3p impaired the proliferation, migration, and invasion of COAD cells and promoted apoptosis, whereas supplementing NEK6 alleviated the damage of the proliferation, migration, and invasion of COAD cells caused by miR-323a-3p and inhibited miR-323a-3p-induced apoptosis. These findings indicate that miR-323a-3p regulates the proliferation, migration, invasion, and apoptosis of COAD cells by targeting NEK6. Conclusion miR-323a-3p downregulates NEK6 in COAD cells; this provides a novel basis for further understanding the occurrence and development of COAD.

Highlights

Colorectal cancer (CRC) is the third most commonly diagnosed cancer and the second leading cause of cancer death worldwide in 2020 [1]
By detecting NEK6 levels in different colon adenocarcinoma (COAD) cell lines, we found that the NEK6 transcription and protein levels in COAD cell lines were higher than those in normal colon epithelial cell lines, and the expression level of NEK6 in SW620 and HCT-8 cells was the highest (p < 0.05; Figures 1(b)–1(d)). erefore, subsequent in vitro experiments were performed using SW620 and HCT-8 cells
Transfection of the miR323a-3p mimic into cells upregulated the expression of miR323a-3p and decreased the expression of NEK6, indicating that miR-323a-3p negatively regulated the expression of NEK6 (p < 0.001; Figures 2(d)–2(f )). e results of the StarBase analysis showed that miR-323a-3p expression was significantly decreased in COAD tissues (Figure 2(g))

Summary

Introduction

Colorectal cancer (CRC) is the third most commonly diagnosed cancer and the second leading cause of cancer death worldwide in 2020 [1]. The incidence of CRC continues to increase every year in many developing countries. E causes of CRC include genetic, environmental, and lifestyle factors. Ese factors may cause mutations or abnormal expression of certain oncogenes and tumor suppressor genes, leading to the occurrence or development of CRC [2]. Erefore, it is necessary to further explore the molecular mechanism of CRC occurrence and development, which can lay the foundation for finding new therapeutic targets and developing novel drugs for CRC. Inhibition of NEK6 can lead to the termination of mitosis, chromatin spindle defects, and abnormal chromosomal differentiation [5–7]. The role and regulatory mechanism of NEK6 on the development of colon adenocarcinoma (COAD) are not fully understood

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Conclusion