PMC8685210_01
CT imaging of changes of masses in oropharynx and left neck. (A) Before chemotherapy (pharyngeal mass: 6.0 cm x 4.6 cm; neck mass: 3.9 cm x 2.9 cm). (B) At the beginning of radiotherapy and 11 days after chemotherapy (pharyngeal mass: 6.0 cm x 4.4 cm; neck mass: 3.5 cm x 2.8 cm). (C) At the end of the radiotherapy and oral crizotinib for 1 month (pharyngeal mass: 4.3 cm x 2.7 cm; neck mass: 2.6 cm x 1.4 cm). (D) At the beginning of oral imatinib and thalidomide (pharyngeal mass: 2.8 cm x 1.9 cm; neck mass: 2.1 cm x 1.4 cm). (E) Oral imatinib and thalidomide for 2 months. (F) Oral imatinib and thalidomide for 6 months.
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PET/CT scan showed a large solid mass located in the left parapharyngeal space with compression of pharyngeal cavity and multiple enlarged lymph nodes in the left neck ( Figure 1A ). CD68 and CD163 and partially express S100 ( Figures 2C-E ). Chemotherapy (cyclophosphamide, doxorubicin, vincristine, prednisone, and etoposide (CHOEP)) was initially started on December 8, 2017 with the aim of stabilization of the fulminate disease course; however, the lesions did not shrink and pain and dysphagia were heavier caused by the compression of the pharynx ( Figure 1B ). Partial response (PR) was observed after radiotherapy of a total dose of 60 Gy/30 F and 1-month treatment of crizotinib, with sum of the product of the longest perpendicular dimensions (SPD) decreased by 57% ( Figure 1C ). The lesions kept shrinking ( Figure 1D ) after radiotherapy, and crizotinib was still taken daily for 1 month more. Figure 1E ) compared with tumor size in March 2018. Four months later the re-evaluation by CT scans showed a nearly complete remission (CR) ( Figure 1F ).
The hematoxylin-eosin (H&E) and immunohistochemical pictures of the tumor. (A) H&E, original magnification, x100. (B) H&E, original magnification, x400. (C) Immunohistochemical staining for CD68. (D) Immunohistochemical staining for CD163. (E) Immunohistochemical staining for S100.
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Figures 2A, B ). For immunohistochemical markers, most HS express CD68 and CD163 and partially express S100 ( Figures 2C-E ). Chemotherapy (cyclophosphamide, doxorubicin, vincristine, prednisone, and etoposide (CHOEP)) was initially started on December 8, 2017 with the aim of stabilization of the fulminate disease course; however, the lesions did not shrink and pain and dysphagia were heavier caused by the compression of the pharynx ( Figure 1B ).
Imatinib could inhibit MAPK and PI3K/AKT pathways in vitro. (A-C) Cells were exposed to different concentrations of imatinib, thalidomide (THD), and imatinib + thalidomide. Cell viability was assessed using CCK8 assays after treatment for 24 h. Data were obtained from three independent experiments. *
p < 0.05, **
p < 0.01, ***
p < 0.001 vs. each control group. (D) KEGG pathway enrichment analysis of gene mutations in patient. (E) Representative immunohistochemistry staining images of p-AKT, p-JNK, and p-ERK in patient's tumor tissues (magnification, x200). (F) Total proteins were collected from the treated DH82 cells, and Western blot analyses for the expression of p-AKT, AKT, p-JNK, JNK, p-ERK, and ERK were performed. GAPDH was used as a loading control. Data were obtained from three independent experiments. ***
p < 0.001 vs. each control group.
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Results of cell counting kit-8 (CCK8) assays showed that the proliferation activity of DH82 was significantly inhibited by imatinib but not thalidomide ( Figures 3A, B ). Combined thalidomide and imatinib treatment did not improve the inhibitory effects of imatinib to DH82 ( Figure 3C ). Further KEGG pathway enrichment analysis of NGS results from patient's tissue also revealed that PI3K/AKT and MAPK pathways were activated in this HS patient ( Figure 3D ). Results showed that p-AKT and p-ERK were strongly positive, while p-JNK was almost negative ( Figure 3E ), indicating the patient actually harbored the activation of MAPK and PI3K/AKT pathways. Treatment of DH82 with imatinib demonstrated that p-ERK and p-AKT were substantially inhibited with imatinib while p-JNK was slightly elevated in a dose-dependent manner, which confirmed the inhibitory effects of imatinib on DH82 by targeting activation of MAPK and PI3K/AKT pathways ( Figure 3F ).