The 3'‑untranslated region of XB130 regulates its mRNA stability and translational efficiency in non‑small cell lung cancer cells

Wang,Qinrong; Liu,Lingling; Gou,Xuanjing; Zhang,Ting; Zhao,Yan; Xie,Yuan; Zhou,Jianjiang; Liu,Ying; Song,Kewei

doi:10.3892/ol.2023.14013

The 3'‑untranslated region of XB130 regulates its mRNA stability and translational efficiency in non‑small cell lung cancer cells

Authors:
- Qinrong Wang
- Lingling Liu
- Xuanjing Gou
- Ting Zhang
- Yan Zhao
- Yuan Xie
- Jianjiang Zhou
- Ying Liu
- Kewei Song
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Affiliations: Key Laboratory of Endemic and Ethnic Diseases, Ministry of Education, Guizhou Medical University, Guiyang, Guizhou 550004, P.R. China
Published online on: August 17, 2023 https://doi.org/10.3892/ol.2023.14013
Article Number: 427
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Silencing XB130 inhibits cell proliferation and epithelial‑mesenchymal transition in non‑small cell lung cancer (NSCLC), suggesting that downregulating XB130 expression may impede NSCLC progression. However, the molecular mechanism underlying the regulation of XB130 expression remains unclear. In the present study, the role of the 3'‑untranslated region (3'‑UTR) in the regulation of XB130 expression was investigated. Recombinant psiCHECK‑2 vectors with wild‑type, truncated, or mutant XB130 3'‑UTR were constructed, and the effects of these insertions on reporter gene expression were examined using a dual‑luciferase reporter assay and reverse transcription‑quantitative PCR. Additionally, candidate proteins that regulated XB130 expression by binding to critical regions of the XB130 3'‑UTR were screened for using an RNA pull‑down assay, followed by mass spectrometry and western blotting. The results revealed that insertion of the entire XB130 3'‑UTR (1,218 bp) enhanced reporter gene expression. Positive regulatory elements were primarily found in nucleotides 113‑989 of the 3'‑UTR, while negative regulatory elements were found in the 1‑112 and 990‑1,218 regions of the 3'‑UTR. Deletion analyses identified nucleotides 113‑230 and 503‑660 of the 3'‑UTR as two major fragments that likely promote XB130 expression by increasing mRNA stability and translation rate. Additionally, a U‑rich element in the 970‑1,053 region of the 3'‑UTR was identified as a negative regulatory element that inhibited XB130 expression by suppressing translation. Furthermore, seven candidate proteins that potentially regulated XB130 expression by binding to the 113‑230, 503‑660, and 970‑1,053 regions of the 3'‑UTR were identified, shedding light on the regulatory mechanism of XB130 expression. Collectively, these results suggested that complex sequence integrations in the mRNA 3'‑UTR variably affected XB130 expression in NSCLC cells.

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