Fig. 2: Expression of C5aR1 and the function of C5aR1 in human PDAC cell lines. | British Journal of Cancer

Fig. 2: Expression of C5aR1 and the function of C5aR1 in human PDAC cell lines.

From: Inhibition of C5a-C5aR1 axis suppresses tumour progression by enhancing antitumour immunity and chemotherapeutic effect in pancreatic ductal adenocarcinoma

Fig. 2

a Western blot analysis of C5a and C5aR1, C5aR2 expression in various human pancreatic cell lines, including cell lines of primary PDAC (BxPC-3, MIA PaCa-2, PANC-1, Capan-2), metastatic PDAC ascites (AsPC-1), lymph node metastasis (Hs766T), and PDAC liver metastases (CFPAC-1, Capan-1). b Western blot analysis of the C5aR1 knockdown with C5aR1siRNA transfection in MIA PaCa-2, BxPC-3 cells. The expression of C5aR1 in C5aR1siRNA1, 2-transfected cells was decreased by normalisation with β-actin compared to the control siRNA cells. c Proliferation assays using C5aR1siRNAs-transfected cells. The absorbance of MIA PaCa-2 and BxPC-3 cells, which had undergone C5aR1 knockdown with C5aR1siRNA1, 2 transfection, showed significant reduction after 48 h in both cells when compared to the control siRNA group (*p < 0.05, **p < 0.01, Mann–Whitney–Wilcoxon test). d The efficacy of C5aR1 selective inhibitor (CCX168) in proliferation assays. The absorbance of MIA PaCa-2 and BxPC-3 cells, which had been seeded with CCX168, showed significant reduction after 48 h in both cells when compared to the control group (*p < 0.05, **p < 0.01, Mann–Whitney–Wilcoxon test). e Western blot analysis with C5aR1siRNA-transfected cells (MIA PaCa-2, BxPC-3) showed decreasing expression of p-PI3K, p-AKT, and p-mTOR by normalisation with β-actin compared to the control cells, whereas the expression of t-PI3K, t-AKT, and t-mTOR showed no difference compared to the control cells. f The analyses of proliferation using Gemcitabine (Gem) and CCX168. The absorbance of MIA PaCa-2 and BxPC-3 cells, which had been seeded with CCX168, showed significant reduction after 48 h in both cells when compared to the control group (*p < 0.05, **p < 0.001, Mann–Whitney–Wilcoxon test). In addition, proliferation in MIA PaCa-2 cells was significantly inhibited after 48 h in the Gem and CCX168 combined group compared to the Gem or CCX168 group. g, h Apoptosis assay utilising annexin V and PI using MIA PaCa-2 cells. The number of early apoptotic cells that were positive for Annexin V, but negative for PI, was significantly increased in the group treated with Gem in combination with C5aR1siRNA transfection or CCX168 group compared to Gem group (*p < 0.05, **p < 0.001, Mann–Whitney–Wilcoxon test).

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