Carboplatin-induced Senescence in Ovarian Cancer Cells Is Reversed Through EGFR and NF-κB Signaling

Abstract

Background: Metastases and recurrence of ovarian cancer after surgery and chemotherapy account for most cancer-related deaths, yet the mechanism underlying metastases and recurrence remains poorly understood. Recent evidence demonstrates that although long-lasting cells were considered tumor suppressors, senescent cancer cells, can induce the metastases and recurrence. In this study, we focused on the fate of ovarian cancer cells treated with carboplatin and explored the mechanism underlying ovarian cancer cell recovery from chemotherapy-induced senescence. Methods: SÁ-β-galactosidase staining was used to detect the impact of carboplatin on senescence of ovarian cancer cells. Cell proliferation was determined using direct cell counting, clone formation assay and 3D tumor spheroid formation assay. Lentivirus-mediated transduction was used to silence or upregulate EGFR expression. Quantitative real-time PCR and western blot analysis validated the efficacy of the knockdown or overexpression effect. Immunofluorescence staining and western blot analysis were used to examined the expression of EGFR and NF-KB. Cell death was determined using trypan blue staining assay. Results: Ovarian cancer cells treated by carboplatin exhibit a senescence-like phenotype indicated by SA-β-galactosidase positive staining. Importantly, carboplatin-induced senescence-like phenotype is reversible. In ovarian cancer cells, EGFR positively regulated cells proliferation, decreased carboplatin-induced senescence and upregulated the NF-κB1 protein level. EGFR/NF-κB1 upregulation promoted the recovery of ovarian cancer cells from senescence and chemoresistance to carboplatin. Conclusions: Ovarian cancer cells treated with carboplatin displayed a reversible senescence-like phenotype that could be combined with EGFR or NF-κB1 inhibitors to improve treatment effects.

Mechanisms of Senescence in Cancer: Positive and Negative Aspects of Cancer Cells Senescence

“…Through SA-β-galactosidase staining in an ovarian cancer cell line, Li et al showed that overexpression of EGFR inhibited the carboplatin-induced senescence-like phenotype. At the same time, EGFR knockdown may contribute to the carboplatin-induced senescence-like phenotype [54]. EGFR knockdown also induced senescence in gallbladder cancer cells GBC-SD and NOZ.…”

The senescent mesothelial matrix accentuates colonization by ovarian cancer cells

“…On the other hand, it has also been shown to potentiate tumor heterogeneity, resistance [6], and relapse [7]. For instance, TIS in ovarian cancer cells leads to their transient dormancy, which can be reversed to re-initiate proliferation [8,9]. TIS-associated cytokine secretion (as part of the broader phenomenon of senescence-associated secretory phenotype or SASP) may accentuate infiltration by tumor-associated macrophages (TAMs) in primary tumors leading to greater invasion and metastasis [10][11][12].…”