Recent studies have revealed that a part of members of DUB are involved in carcinogenesis such as USP1, USP7 and USP2 (25–27) while some DUB inhibited various cancer progression such as BAP1 and USP10 (28, 29). USP 42, a DUB related to p53 in addition to histone H2B has been discovered to be involved in multiple cancers (30, 31). Misaki Matsui, et al. revealed that USP42 played important role in DSB repair mediated by nuclear speckle by accelerating BRCA1 to load to DSB sites (32). Amélie Giguère et al. found that RUNX1 rearrangements combined USP42 played an essential part in acute myeloid leukemia, predicting a promising therapeutic target (33). Nevertheless, whether and how USP42 exerts crucial roles in breast cancer progress are not elucidated at all. In our present study, we uncovered for the first time that breast cancer tissues expressed higher load of USP42 at both mRNA and protein level compared to normal tissue samples. Moreover, expression level of USP42 was correlated with survival time of patients with breast cancer. Our primary findings provided compelling evidence for indispensable effects of USP42 on breast cancer.
Given these data, we assumed that USP42 served as an oncogene during the development of breast cancer. To confirm this hypothesis, we constructed plasmid with overexpressing USP42 and transfecting it into breast cancer cells. Comparing with control group, overexpressing USP42 dramatically promoted breast cancer cells growth, invasion and migration. Subsequently, we employed RNAi technology, the common strategy utilized in cancer research, to silence the expression of USP42 effectively in breast cancer cells. Knockdown of USP42 significantly suppressed breast cancer cells proliferation, invasion and migration. Taken together, USP42 was capable of facilitating the progression of breast cancer.
Apoptosis has been proved to be a well-organized and intricate process in various biological conditions and it exerts crucial effects in multiple diseases (34). One of the leading reasons for a variety of cancers occurrence is that too little apoptosis produces and tumor cells will not disappear (35, 36). In this study, we employed flow cytometry to confirm that silencing USP42 significantly increased apoptosis rate of breast cancer cell. In addition, Bcl-2, the member of Bcl-2 family, is the main regulator of apoptosis. Increased ration of Bax/Bcl-2 in addition to caspase-3 activation effectively triggered apoptosis and cell death (37). In this study, we investigated the load of apoptosis-related proteins including Bcl-2, Bax and cleaved-caspase-3, we found that silencing USP42 notably iecreased the expression of Bax in addition to cleaved-caspase-3 while decreased the expression of Bcl-2, indicating that USP42 promoted breast cancer cells progression by mediating cell apoptosis.
Previous studies have uncovered that MAPK signaling pathway is involved in various physiological progress such as cell proliferation and apoptosis (38). JNKs and p38 kinases are two indispensable subfamilies of MAPK signaling pathway and the activation JNKs as well as p38 kinases responds to multiple endogenous and exogenous stimuli (38, 39). Study has discovered that phosphorylation of JNK and p38 is associated with apoptosis tightly (37). In our present study, we revealed that knockdown of USP42 obviously up-regulated phosphorylation of JNK and p38 while JNK inhibitor or p38 inhibitor partly eliminated the above activation, which implied that silencing USP42 accelerated breast cancer cell apoptosis by activating JNK/p38 signaling pathway. Further investigation verified that pretreatment with JNK inhibitor of p38 inhibitor partially counteracted effects of silencing USP42 on breast cancer cells progression: increased cell viability, inhibited invasion in addition to migration and depressed apoptosis. However, we need more experiments to uncover that how USP42 affect the activation of JNK as well as p38. Additionally, we need to explore the direct target of USP42 on MAPK pathway and confirm the accurate deubiquitination sites.
In conclusion, our findings firstly expounded the functions of USP42 on breast cancer cells and revealed its mechanisms for the first time, which meant that USP42 may be regarded as a potential gene target for treating breast cancer.