PURPOSE: Radioresistance is a major cause of treatment failure in tumor radiotherapy and the underlying mechanisms of radioresistance are still elusive. Golgi phosphoprotein 3 (GOLPH3) has been reported to associate tightly with cancer progression and chemoresistance. Herein, we explored whether GOLPH3 mediated radioresistance of lung adenocarcinoma (LUAD) and whether targeted suppression of GOLPH3 sensitized LUAD to radiotherapy.
METHODS AND MATERIALS: The aberrant expression of GOLPH3 was evaluated by immunohistochemistry in LUAD clinical samples. To evaluate the association between GOLPH3 and radioresistance, colony formation and apoptosis were assessed in control and GOLPH3 knockdown cells. γ-H2AX foci/level determination and micronucleus test were used to analyze DNA damage production and repair. The rescue of GOLPH3 knockdown was then performed by exogenous expression of siRNA-resistant mutant GOLPH3 to confirm the role of GOLPH3 in DNA damage repair. Mechanistically, the effect of GOLPH3 on regulating stability and nuclear accumulation of epidermal growth factor receptor (EGFR) and the activation of DNA-PK were investigated by qRT-PCR, western blot, immunofluorescence and co-immunoprecipitation. The role of GOLPH3 in vivo in radioresistance was determined in a xenograft model.
RESULTS: In tumor tissues of 33 patients with LUAD, the expression of GOLPH3 showed significantly increases compared with those in matched normal tissues. Knocking down GOLPH3 reduced the clonogenic capacity, impaired DSB repair and enhanced apoptosis after irradiation. In contrast, reversal of GOLPH3 depletion rescued the impaired repair of radiation-induced DSBs. Mechanistically, loss of GOLPH3 accelerated the degradation of EGFR in lysosome, causing the reduction in EGFR levels, thereby weakening nuclear accumulation of EGFR and attenuating the activation of DNA-PK. Furthermore, adenovirus-mediated GOLPH3 knockdown could enhance the ionizing-radiation response in LUAD xenograft model.
CONCLUSIONS: GOLPH3 conferred resistance of LUAD to ionizing-radiation via stabilizing EGFR and targeted suppression of GOLPH3 might be considered as a potential therapeutic strategy for sensitizing LUAD to radiotherapy.
|Journal||International journal of radiation oncology, biology, physics|
|Early online date||26 Nov 2021|
|Publication status||Early online date - 26 Nov 2021|