Fig 2: OTUD5 affected the resistance of NSCLC cells to cisplatin and doxorubicin. (a–d) The expression of OTUD5 in A549 and HCC827 cells treated with cisplatin and doxorubicin was detected by Western blot and RT‐PCR. **p < .01 and ***p < .001. (e) After cisplatin and doxorubicin treatments, the CCK‐8 assay was performed to detect the viability of A549 and HCC827 cells transfected with OTUD5 siRNA or NC siRNA. *p < .05, **p < .01, and ***p < .001. (f) The IC50 values of cisplatin and doxorubicin in NSCLC cells were compared before and after A549 and HCC827 cells were transfected with OTUD5 siRNA or NC siRNA. **p < .01 and ***p < .001. After cisplatin and doxorubicin treatments, flow cytometry analysis was used to detect the apoptosis of A549 (g) and HCC827 (h) cells transfected with OTUD5 siRNA or NC siRNA. **p < .01 and ***p < .001 [Colour figure can be viewed at wileyonlinelibrary.com]

Fig 3: Knockdown of OTUD5 promoted the proliferation, migration, and invasion of HCC827 cells while inhibiting their apoptosis via regulating PDCD5. Western blot (a) and RT‐PCR (b) were used to detect PDCD5 expression in HCC827 cells transfected with OTUD5 siRNA and NC siRNA. CCK‐8 assay, transwell assay, and apoptosis analysis were carried out to detect the proliferation (c), migration (d), invasion (e), and apoptosis (f) of HCC827 cells, respectively. *p < .05, **p < .01, and ***p < .001 [Colour figure can be viewed at wileyonlinelibrary.com]

Fig 4: The expression of OTUD5 was reduced in NSCLC. The Kaplan–Meier method was used to analyze the survival time of patients with lung adenocarcinoma (a), pancreatic cancer (b), and cervical squamous carcinoma (c) who showed high or low expression of OTUD5. (d–e) RT‐PCR and Western blot were applied to detect the expression of OTUD5 mRNA and protein in tumor tissues and adjacent normal tissues, respectively. ***p < .001. (f–g) RT‐PCR and Western blot were applied to detect the expression of OTUD5 mRNA and protein in NSCLC cell lines A549, H358, H1299, and HCC827, and human bronchial epithelial cell line BEAS‐2B. ***p < .001 [Colour figure can be viewed at wileyonlinelibrary.com]

Fig 5: OTUD5 impeded the progression of NSCLC cells via regulating p53. (a) Western blot was applied to measure p53 expression in A549 cells transfected with OTUD5 siRNA and NC siRNA. (b) RT‐PCR was applied to measure p53 mRNA expression in A549 cells transfected with OTUD5 siRNA and NC siRNA. CCK‐8 assay, transwell assay, and apoptosis analysis were carried out to detect the proliferation (c), migration (d), invasion (e), and apoptosis (f) of A549 cells, respectively. *p < .05, **p < .01, and ***p < .001. (g) The Pearson correlation test was used to analyze the correlation between OTUD5 and p53 expression in NSCLC samples [Colour figure can be viewed at wileyonlinelibrary.com]