Multi-SUMOylation of NAC1 is essential for the growth of prostate cancer cells.

Nucleus accumbens-associated 1 (NAC1) is a member of pox virus and zinc finger/bric-a-brac tramtrack broad complex (BTB/POZ) gene family. Overexpression of NAC1 is implicated in cancer development, recurrence and chemotherapy resistance. In our previous study, we found NAC1 was a potential small ubiquitin-like modifier (SUMO) substrate in prostate cancer cells. However, there was still lack of evidences to further support and validate the result. In this work, we found that NAC1 is a multi-SUMO-sites acceptor. The SUMO acceptor lysines were K167, K318, K368, K483 and K498. SUMOylation didn't alter the localization of NAC1, but facilitated the formation of NAC1 nuclear bodies. Compared with NAC1 wild type (NAC1 WT), the SUMO-sites mutant of NAC1 (NAC1 SM) suppressed cell proliferation and tumor growth in cellular and animal levels. This work uncovered the function of SUMOylation of NAC1 in prostate cancer cells.

Nucleus accumbens associated 1 is recruited within the promyelocytic leukemia nuclear body through SUMO modification.

Nucleus accumbens associated 1 (NACC1) is a cancer-associated BTB/POZ (pox virus and zinc finger/bric-a-brac tramtrack broad complex) gene, and is involved in several cellular functions in neurons, cancer and stem cells. Some of the BTB/POZ proteins associated with cancer biology are SUMOylated, which appears to play an important role in transcription regulation. We show that NACC1 is SUMOylated on a phylogenetically conserved lysine (K167) out of three consensus SUMOylation motif sites. Amino acid substitution in the SIM sequence (SIM/M) within the BTB/POZ domain partially reduced K167 SUMOylation activity of NACC1. Overexpression of GFP-NACC1 fusion protein leads to formation of discrete nuclear foci similar to promyelocytic leukemia nuclear bodies (PML-NB), which colocalized with SUMO paralogues (SUMO1/2/3). Both NACC1 nuclear body formation and colocalization with SUMO paralogues were completely suppressed in the GFP-NACC1-SIM/M mutant, whereas they were partially maintained in the NACC1 K167R mutant. Confocal immunofluorescence analysis showed that endogenous and exogenous NACC1 proteins colocalized with endogenous PML protein. A pull-down assay revealed that the consensus motifs of the SUMO acceptor site at K167 and the SIM within the BTB/POZ domain were both necessary for efficient binding to PML protein. Our study demonstrates that NACC1 can be modified by SUMO paralogues, and cooperates with PML protein.

Clinical significance of miR-1298 in cervical cancer and its biological function in vitro.

Cervical cancer is one of the most malignant tumors in women. miR-1298 was reported to be abnormally expressed and serve crucial role in tumorigenesis of several types of cancer; however, the role of miR-1298 in cervical cancer remains unknown. The present study aimed to evaluate the clinical and biological significance of miR-1298 in cervical cancer. To do so, the expression level of miR-1298 in cervical cancer tissues and cells was evaluated by reverse transcription quantitative PCR. Kaplan-Meier survival analysis and Cox regression analysis were used to explore the prognostic significance of miR-1298 in patients with cervical cancer. Cell Counting Kit-8 and Transwell migration and invasion assays were used to evaluate the effect of miR-1298 on the proliferative, migratory and invasive abilities of cervical cancer cells, respectively. The expression of miR-1298 was lower in cancer tissues and cells compared with normal tissues and cells. Furthermore, miR-1298 expression was associated with lymph node metastasis, tumor diameter and staging from the International Federation of Gynecology and Obstetrics. In addition, patients with low miR-1298 expression had poorer overall survival. These findings suggested that miR-1298 may be considered as an independent prognostic factor for patients with cervical cancer. Furthermore, the results demonstrated that miR-1298 knockdown could promote tumor cell proliferation and migratory and invasive abilities. In addition, nucleus accumbens-associated 1 (NACC1) was demonstrated to be a direct target of miR-1298. Taken together, these findings indicated that miR-1298 overexpression may be considered as a prognostic biomarker for cervical cancer and that miR-1298 may play an inhibitor role in cervical cancer by targeting NACC1.