Senescence-associated secretory factors induced by cisplatin in melanoma cells promote non-senescent melanoma cell growth through activation of the ERK1/2-RSK1 pathway

Abstract

Although targeted therapy and immunotherapy greatly improve the outcome of melanoma, drug resistance and low response rates still maintain the unsubstitutability of traditional chemotherapy. Cisplatin (CDDP) is widely used in different types of tumours with high response rates, but it generally has low efficiency in melanoma. The mechanisms underpinning the phenomena are not sufficiently understood. Here we demonstrated that various melanoma cell lines adopted senescence phenotype after CDDP treatment in contrast to the other types of tumour cells. CDDP treatment induced melanoma A375 cells into senescence through the sequential activation of the DNA damage response and the P53/P21 pathway. All the senescent melanoma cells induced by CDDP alone or the combination of CDDP and dacarbazine developed robust senescence-associated secretory phenotype (SASP), that is, the secretion of multiple cytokines. IL-1α was an early component and an upstream regulator of SASP. Similarly, CDDP either alone or combined with dacarbazine could induce melanoma cell senescence and SASP in either A375 or B16F10 melanoma xenograft mice. The supernatant of senescent A375 cells promoted the growth of normal non-senescent A375 cells and enhanced their expression and secretion of IL-8 through the activation of the ERK1/2-RSK1 pathway. The transplantation of non-senescent and senescent A375 cells together into nude mice showed accelerated tumour growth compared with transplanting non-senescent cells alone; no tumours developed when transplanting senescent cells alone. Following CDDP administration in A375-bearing mice, the intratumour injection of neutralisation antibodies targeting the SASP factors IL-1α or IL-8 evidently delayed tumour growth. The results suggest that the CDDP-induced senescent melanoma cells promote non-senescent cells proliferation through the activation of ERK1/2-RSK1 pathway by the SASP factors. Cell senescence and concomitant SASP may be the particular mechanisms for melanoma to resist chemotherapeutics.

Therapy-Induced Senescence: An “Old” Friend Becomes the Enemy

“…Reduced DNA synthesis, morphology, SA-β-gal, growth arrest, p21 Cip1 [55] Irinotecan SGC-7901, MKN-45 SA-β-gal [56] A549, HCT116 Reduced DNA synthesis, morphology, SA-β-gal, growth arrest, p21 Cip1 [55] Alkylating agents Busulfan Rat-derived BMSCs and ADSCs SA-β-gal [42] WI38 Growth arrest, SA-β-gal [57] U2OS, MG63 SA-β-gal [58] WI38 SA-β-gal [59] Murine hematopoietic cells SA-β-gal, p16 INK4 , p19 INK4 [60] Temozolomide Patient derived glioma cells Cell cycle arrest, polyploidy, morphology [61] GL261 SAHF (H3K9Me3), p53, Rb [62] LN229 SA-β-gal, cell cycle arrest, SASP (IL-6, IL-8) [63] In vivo (p16-3MR) mouse model p16 INK4 [28] Carmustine GL261 SAHF (H3K9Me3), p53, Rb [62] Dacarbazine A375, B16F10 SASP [64] Cyclophosphamide HSC-bcl2 lymphoma SA-β-gal, p53, p16 INK4 [65] Melphalan Multiple myeloma mouse model SA-β-gal [66] Mitomycin C A549 Growth arrest, SA-β-gal, yH2AX, morphology [67]…”

“…Platinum-based Cisplatin A375, B16F10, B16F10 xenografts SASP, SA-β-gal [64] A2780 SAHF (HP1-γ), morphology, SA-β-gal [68] CNE1 Growth arrest, morphology, SA-β-gal [30] SKOV3, TOV-21G Morphology, SA-β-gal [69] HepG2, SMMC-7721 SA-β-gal, p53, p21 Cip1 , p16 INK4 [70] Follicular lymphoma 3D model SA-β-gal [49] In vivo mouse model (p16-3MR) p16 INK4 [28] Carboplatin H1299, patients' lung tumor samples Cell cycle arrest, SA-β-gal, p16 INK4 , RB, downregulation of cyclin B1 and cyclin D1 [71] Oxaliplatin PROb, CT26 SA-β-gal [72] HepG2, SMMC-7721, patients' colorectal tumor samples SA-β-gal [73] Antimetabolites Methotrexate C85 p53 [74] C85 SA-β-gal [75] MCF-7 SA-β-gal [76] Rat-derived BMSCs and ADSCs SA-β-gal [42] A549 p21 Cip1 , low Ki67, growth arrest, SA-β-gal, increased granularity, morphology, SASP (IL-6, IL-8), γH2AX [43] SH-SY-5Y p21 Cip1 , growth arrest, SA-β-gal, γH2AX [43] HCT116 p21 Cip1 , low Ki67, growth arrest, SA-β-gal, increased granularity, morphology, SASP (IL-8), γH2AX [43] MDA-MB-231 p21 Cip1 , growth arrest, SA-β-gal, morphology, SASP (IL-6, IL-8, VEGF) [43] MCF-7 p21 Cip1 , low Ki67, growth arrest, SA-β-gal, increased granularity, morphology, γH2AX [43] Pemetrexed H1650, A549, H2228, H292, H226 and H1650, A549 xenografts SA-β-gal, morphology, SASP (IL-6, IL-8, IL-1β and MCP-1) [77] A549 SASP, SA-β-gal [78] Gemcitabine Miapaca-2 and Panc-1 SA-β-gal [79] AsPc1, Panc1 SA-β-gal [80] Azacitidine U2OS, MCF7 SA-β-gal, p53, growth arrest [81] TPC-1 SA-β-gal [82] KKU100, HuCCA1, RMCCA1 Morphology, SA-β-gal [83] DU145, LNCaP Morphology, growth arrest, polyploidy [40] Bromodeoxyuridine MNA, STA-NB-10, CLB-Ma mouse xenograft (MYCN-amplified neuroblastoma)…”

“…Other alkylating agents, such as carmustine, dacarbazine, cyclophosphamide, and melphalan have also been shown to induce senescence in a variety of cell lines [62,64]. Dacarbazine, in combination with cisplatin, induced robust SASP in A375 and B16F10 cell lines, despite the absence of significant SA-β-gal expression [64]. Moreover, both melphalan and cyclophosphamide were able to induce senescence in multiple myeloma and lymphoma mouse models, respectively [65,66].…”

“…Cisplatin was the first compound tested for the induction of TIS in tumor cells. Cisplatin induces senescence in several melanoma cell lines associated with a vigorous secretory response [64]. Cisplatin also induces senescence in A2780, SKOV3 and TOV-21G ovarian cancer cells; A549 and H292 lung cancer cells; HepG2 and SMMC-7721 hepatocellular cancer cells and CNE1 nasopharyngeal cancer cells [30,[68][69][70].…”

Stimulation of prostate cells by the senescence phenotype of epithelial and stromal cells: Implication for benign prostate hyperplasia

“…Mitogen‐activated serine/threonine kinases ERK1 and ERK2 promote cell proliferation and survival although, under certain conditions, they may induce apoptosis . We should note that ERK1/2 activation due to DNA damage induced cellular senescence was also observed for cisplatin‐treated melanoma cells . The longer culture time required to activate AKT and ERK compared to STAT5 (9 hours vs 6 hours, Figure ) may be due to delayed accumulation of the factor(s) that direct AKT and ERK activation.…”