CHD1L accelated the progression of cutaneous squamous cell carcinoma via promoting PI3K/PD-L1 signaling pathway induced M2 polarization of TAMs

Abstract

To investigate CHD1L's impacts and molecular processes in hypoxic cutaneous squamous cell carcinoma. Monoclonal proliferation assays and CCK-8 were used to detect the proliferation capacity of A431 cells and Colon16 cells; wound healing experiments and Transwell assays were used to examine the migration and invasion capacity of A431 cells and Colon16 cells; angiogenesis experiments were conducted to assess the influence of A431 cells on angiogenesis; a nude mouse tumor xenograft experiment and HE staining were utilized to evaluate the impact of CHD1L on the progression of cutaneous squamous cell carcinoma; western blot analysis was performed to detect the expression of p-PI3K, p-AKT, and PD-L1 in A431 cells, as well as CD9, TSG101, PD-L1 in exosomes, and CD206, Arginase-1, iNOS, IL-1β, p-AKT, p-mTOR, VEGF, COX-2, MMP2, MMP9, p-ERK1/2 in tumor-associated macrophages. Under hypoxic conditions, CHD1L promoted the proliferation, migration, invasion, and angiogenesis of cutaneous squamous cell carcinoma. Furthermore, CHD1L facilitated the progression of cutaneous squamous cell carcinoma. CHD1L also increased the relative protein expression of p-PI3K, p-AKT, and PD-L1 in A431 cells, as well as CD9, TSG101, PD-L1 in exosomes, CD206, Arginase-1, p-AKT, p-mTOR, VEGF, COX-2, MMP2, MMP9, and p-ERK1/2 in tumor-associated macrophages, while inhibiting the relative protein expression of iNOS and IL-1β. Under hypoxic conditions, CHD1L can promote the proliferation and migration of cutaneous squamous cell carcinoma.

Keywords: CHD1L; Cutaneous squamous cell carcinoma; Hypoxia; Tumor-associated macrophages.

Fig. 1

CHD1L promotes the progression of SCC. (A) H&E staining results; (B) Tumor area statistics. Data are expressed as mean ± standard deviation; N = 6; **P < 0.01.

Fig. 2

CHD1L promotes the proliferation of cutaneous squamous cell carcinoma cells in a hypoxic environment. (A) Clonal proliferation assay results for A431 and Colon16 cells; (B) Plot of statistical results of clone formation in A431 cells and Colon16 cells; (C) Results of CCK-8 assay on A431 cells and Colon16 cells. Data are expressed as mean ± standard deviation; N = 3; **P < 0.01.

Fig. 3

CHD1L promotes the migration and invasion of cutaneous squamous cell carcinoma cells in a hypoxic environment. (A) Results of 0 h and 48 h wound healing tests with A431 cells and Colon16 cells; (B) Wound closure rate statistics for A431 cells and Colon16 cells; (C) Results of Transwell assay of A431 and Colon16 cells; D: Statistics on the amount of cell migration and invasion in A431 and Colon16 cells. Data are expressed as mean ± standard deviation; N = 3; **P < 0.01.

Fig. 4

CHD1L promotes angiogenesis of cutaneous squamous cell carcinoma cells in a hypoxic environment. (A) Angiogenesis assay results; (B) Statistics of new blood vessel formation. Data are expressed as mean ± standard deviation; N = 3; **P < 0.01.

Fig. 5

CHD1L promotes the expression of PD-L1 in cutaneous squamous cell carcinoma cells and promotes M2 polarization in TAMs. (A) p-PI3K, p-AKT, and PD-L1 protein bands and relative protein expression levels in A431 cells; (B) CD206, Arginase-1, iNOS, IL-1β, p-AKT, p-mTOR, VEGF, COX-2, MMP2, MMP9, and p-ERK1/2 protein bands and relative protein expression levels in tumor-associated macrophages. GAPDH as control protein; Data are expressed as mean ± standard deviation; N = 6; **P < 0.01.

Fig. 6

CHD1L promotes the expression of PD-L1 in cutaneous squamous cell carcinoma cells in a hypoxic environment and promotes M2 polarization in TAMs through exosomes. (A) p-PI3K, p-AKT, and PD-L1 protein bands and relative protein expression levels in A431 cells; (B) CD9, TSG101, and PD-L1 protein bands and protein expression levels in exosomes; (C) CD206, Arginase-1, iNOS, IL-1β, p-AKT, p-mTOR, VEGF, COX-2, MMP2, MMP9, and p-ERK1/2 protein bands and relative protein expression levels in tumor-associated macrophages. GAPDH as control protein; Data are expressed as mean ± standard deviation; N = 3; **P < 0.01.

Fig. 7

In the hypoxic environment of cutaneous squamous cell carcinoma, CHD1L can promote the secretion of PD-L1 by cancer cells, thus promoting the activation of the ERK/AKT/mTOR signaling pathway and M2 polarization in tumor-associated macrophages, inhibiting M1 polarization and leading to the proliferation, migration, invasion, and angiogenesis of cutaneous squamous cell carcinoma cells.