USP3 promotes proliferation of non-small cell lung cancer through regulating RBM4
Authors: Y Wang
X-H Liao
B Zhong
S-Y Zhu
Journal: European review for medical and pharmacological sciences (2020)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: USP3; non-small cell lung cancer; RBM4; proliferation; qRT-PCR
Affiliations: Department of Respiratory Medicine, The First Affiliated Hospital of Gannan Medical University, Ganzhou, China. .

Abstract:  Objective: Previous studies have shown that ubiquitin specific protease 3 (USP3) is an oncogene. However, the role of USP3 in non-small cell lung cancer (NSCLC) has not been reported. This study aims to explore the expression characteristics of USP3 in NSCLC, and its regulation on the proliferative capacity of NSCLC cells. Patients and methods: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine the expression levels of USP3 and RNA Binding Motif 4 (RBM4) in 42 pairs of tumor tissues and adjacent tissue specimens collected from NSCLC patients. Meanwhile, the correlation between the messenger ribonucleic acid (mRNA) expressions of USP3 and RBM4, and the clinical indicators and prognosis of NSCLC patients were analyzed. At the same time, mRNA expression of USP3 in NSCLC cell lines was further verified by the qRT-PCR method. In addition, USP3 knockdown and overexpression models were constructed using lentivirus in NSCLC cell lines H1299 and SPCA1. Cell counting kit-8 (CCK-8), cell colony formation, and 5-Ethynyl-2'-deoxyuridine (EDU) assays were performed to evaluate the influence of USP3 on proliferative capacity in NSCLC cells. Finally, Dual-Luciferase reporter assay and rescue experiments were conducted to further explore its underlying molecular mechanism. Results: In this experiment, qRT-PCR results revealed that the expression level of USP3 in tumor tissues of NSCLC patients was remarkably higher than that in adjacent tissues, and the difference was statistically significant. Compared with NSCLC patients with low expression of USP3, those with high USP3 expression suffered much more advanced pathology stage and lower overall survival rate. Proliferation ability of NSCLC cells overexpressing USP3 was remarkably enhanced, while the opposite result was observed in the USP3 knockdown group. Subsequently, RBM4 expression in NSCLC tissue samples was found to be significantly reduced and negatively correlated with USP3 level. In addition, the result of Dual-Luciferase reporter assay demonstrated that USP3 can be targeted by RBM4. Rescue experiments revealed that RBM4 was responsible for NSCLC progression regulated by USP3. Conclusions: The above studies indicated that USP3 expression was remarkably up-regulated in NSCLC tissues, which was closely related to the pathological staging and poor prognosis of NSCLC patients. Therefore, USP3 might accelerate the proliferation of NSCLC cells via regulating RBM4. 

Retracted: No

PubPeer comments:
  The investigation was approved by the Ethics Committee of the hospital, and all patients had signed informed consent. This is unconvincing, as the investigation was supposedly distributed over **two** hospitals - The First Affiliated Hospital of Gannan Medical University, Ganzhou, and Fuxin Central Hospital, Fuxin. Fig 1C:  Kaplan-Meier survival curve of NSCLC patients based on USP3 expression. The prognosis of patients with high expression of USP3 was significantly worse than that of low expression group . ![file] Versions of this pair of K-M plots (resembling combs, or walls made of Lego) appear in a great number of papers in this journal. The precipitous nose-dive to 0% Survival at 75 months is especially remarkable and I hope that it doesn t reflect a widespread policy of euthanasia. , 
 Another recurrent feature of this tradition of papers is a Table 1 in which the column heads promise to convert the numbers in each patient subgroup into percentages, but the promise remains unfulfilled. ![file] , 
 A colony-formation plate is re-used between Figs 2C and 4C,  Colony formation assay was performed to detect relative colony number in H1299 and SPCA1 cell lines ...  Colony formation assay was performed to detect the number of colonies in co-transfected cells . ![file] ,