CD81 Enhances Radioresistance of Glioblastoma by Promoting Nuclear Translocation of Rad51

SIMPLE SUMMARY: CD81 is highly expressed in glioblastoma (GBM) as a transmembrane protein. The functional study demonstrated that CD81 contributed to radioresistance of GBM. Further evidence showed that CD81 expression was closely related to DNA damage response and homologous recombination repair (HRR) was responsible for the CD81 mediated radioresistance. Particularly, nuclear membrane protein CD81 assisted the nuclear transport of Rad51, a key protein involved in HRR process after irradiation. Overall, CD81 may be utility as a predictive biomarker and therapeutic target of radioresistant GBM. ABSTRACT: Glioblastoma (GBM) is the most common type of primary tumor in central nervous system in adult with a 5-year survival rate of ≤5%. Despite of recent advances in tumor radiotherapy, the prognosis of GBM remains to be dismal due to radioresistance. In this study, we identified CD81 as a potential biomarker of GBM radioresistance with the analysis of upregulated genes in human glioma radioresistant cell lines U251R and T98G in comparison with U251 cells. In vitro and in vivo experiments demonstrated that suppressing CD81 by siRNA/shRNA enhanced radiation-induced cell killing and DNA damage of γ-H2AX formation, and delayed tumor xenograft growth of GBM. Mechanistically, we found that knockdown of CD81 significantly decreased radiation-induced expression of nuclear Rad51, a key protein involved in homologous recombination repair (HRR) of DNA, suggesting that CD81 is essential for DNA damage response. Meanwhile, when the cells were treated with B02, a Rad51 inhibitor, silencing CD81 would not sensitize GBM cells to radiation, which further illustrates that Rad51 acts as an effector protein of CD81 in tumor radioresistance. Dual immunofluorescence staining of CD81 and Rad51 illustrated that nuclear membrane CD81 contributed to the nuclear transport of Rad51 after irradiation. In conclusion, we demonstrated for the first time that CD81 not only played a vital role in DNA repair through regulating Rad51 nuclear transport, but also might serve as a potential target of GBM radiotherapy.