Dysregulation of CUL4A and CUL4B Ubiquitin Ligases in Lung Cancer

Abstract

The Cullin-RING ubiquitin ligase 4 (CRL4) is implicated in controlling cell cycle, DNA damage repair, and checkpoint response based on studies employing cell lines and mouse models. CRL4 proteins, including CUL4A and CUL4B, are often highly accumulated in human malignancies. Elevated CRL4 attenuates DNA damage repair and increases genome instability that is believed to facilitate tumorigenesis. However, this has yet to be evaluated in human patients with cancer. In our study, 352 lung cancer and 62 normal lung specimens of Asian origin were constructed into tissue microarrays of four distinct lung cancer subtypes. Expression of CUL4A, CUL4B, and their substrates was detected by immunohistochemistry and analyzed statistically for their prognostic value and association with DNA damage response and genomic instability. Our results show that both CUL4A and CUL4B are overexpressed in the majority of lung carcinomas (P_CUL4A <0.001 and P_CUL4B <0.001) and significantly associated with tumor size (P_CUL4A <0.001 and P_CUL4B = 0.002), lymphatic invasion (P_CUL4A = 0.004 and P_CUL4B <0.001), metastasis (P_CUL4A = 0.019 and P_CUL4B = 0.006), and advanced TNM stage (P_CUL4A <0.001 and P_CUL4B <0.001), which parallels gene amplification and abnormal activation of the canonical WNT signaling. Moreover, overexpression of CUL4A, but not CUL4B, is significantly associated with tobacco smoking (p = 0.01) and is inversely correlated with XPC and P21, both of which are substrates of CUL4A (P_CUL4A = 0.019 and P_CUL4B = 0.006). Higher levels of CUL4A or CUL4B are significantly associated with the overall survival of patients (P_CUL4A <0.001 and P_CUL4B <0.001) and progression-free survival (P_CUL4A <0.001 and P_CUL4B = 0.001). Our findings revealed that CUL4A and CUL4B are differentially associated with etiologic factors for pulmonary malignancies and are independent prognostic markers for the survival of distinct lung cancer subtypes.

Keywords: E3 ubiquitin ligase; cell signaling; immunohistochemistry; lung cancer; protein degradation; ubiquitin; ubiquitin ligase.