Elaiophylin Elicits Robust Anti-Tumor Responses via Apoptosis Induction and Attenuation of Proliferation, Migration, Invasion, and Angiogenesis in Pancreatic Cancer Cells

Abstract

Pancreatic cancer remains a formidable challenge in oncology due to its aggressive nature and limited treatment options. In this study, we investigate the potential therapeutic efficacy of elaiophylin, a novel compound, in targeting BxPC-3 and PANC-1 pancreatic cancer cells. We comprehensively explore elaiophylin's impact on apoptosis induction, proliferation inhibition, migration suppression, invasion attenuation, and angiogenesis inhibition, key processes contributing to cancer progression and metastasis. The results demonstrate that elaiophylin exerts potent pro-apoptotic effects, inducing a substantial increase in apoptotic cells. Additionally, elaiophylin significantly inhibits proliferation, migration, and invasion of BxPC-3 and PANC-1 cells. Furthermore, elaiophylin exhibits remarkable anti-angiogenic activity, effectively disrupting tube formation in HUVECs. Moreover, elaiophylin significantly inhibits the Wnt/β-Catenin signaling pathway. Our findings collectively demonstrate the multifaceted potential of elaiophylin as a promising therapeutic agent against pancreatic cancer via inhibition of the Wnt/β-Catenin signaling pathway. By targeting diverse cellular processes crucial for cancer progression, elaiophylin emerges as a prospective candidate for future targeted therapies. Further investigation of the in vivo efficacy of elaiophylin is warranted, potentially paving the way for novel and effective treatment approaches in pancreatic cancer management.

Keywords: BxPC-3; HUVECs; PANC-1; Wnt/β-Catenin; elaiophylin; pancreatic cancer.

Figure 1

The chemical structure of elaiophylin.

Figure 2

Elaiophylin inhibits the proliferation of BxPC-3 and PANC-1 cells. (A) The bar chart indicates the cell viability of BxPC-3 cells treated with elaiophylin at indicated concentrations. (B) The bar chart indicates the cell viability of PANC-1 cells treated with elaiophylin at indicated concentrations. (C,E) Representative images of colony formation of BxPC-3 and PANC-1 cells treated with elaiophylin at indicated concentrations. (D,F) The bar chart indicates the colony-formation rate. (G) Representative western blotting images of PCNA, HDAC1, and β-actin in BxPC-3 cells treated with elaiophylin at indicated concentrations. The original western blotting images are shown in Figures S2 and S3. (H,I) Bar charts indicate the ratio of PCNA/β-actin and HDAC1/β-actin. Error bars, S.D.: * p < 0.05; ** p < 0.01; *** p < 0.001. Data are presented as mean ± SD from three independent experiments (n = 3).

Figure 3

Elaiophylin promotes the cell-cycle arrest of BxPC-3 cells. (A) Representative flow cytometry images showing the cell cycle of elaiophylin-treated BxPC-3 cells stained with PI reagent. The peaks in the FACS graph represent cells in the G0/G1, S, and G2/M phases of the cell cycle. The shaded area indicates the cell population with specific DNA content, representing cells in different phases of the cell cycle. (B) The bar chart indicates the cell-cycle distribution rate. Error bars, S.D.: ** p < 0.01; *** p < 0.001. Data are presented as mean ± SD from three independent experiments (n = 3).

Figure 4

Elaiophylin induces cell apoptosis in BxPC-3 cells. (A) Representative flow cytometry images showing the cell apoptosis of elaiophylin-treated BxPC-3 cells stained with Annexin V-FITC and PI reagent. (B) The bar charts indicate the cell apoptotic rate. (C) Representative western blotting images of Caspase-3, Cleaved-caspase-3, and β-actin in BxPC-3 cells treated with elaiophylin at indicated concentrations. The original western blotting images are shown in Figure S4. (D) The bar charts indicate the ratio of Cleaved-caspase-3/caspase-3. Error bars, S.D.: * p < 0.05; ** p < 0.01; *** p < 0.001. Data are presented as mean ± SD from three independent experiments (n = 3).

Figure 5

Elaiophylin inhibits the migration and invasion of BxPC-3 and PANC-1 cells. (A,C) Representative cell scratch images of BxPC-3 and PANC-1 cells treated with elaiophylin at indicated concentrations. Magnification: 10×, scale bar: 100 µm. (B,D) The bar chart indicates the wound-closure rates of BxPC-3 and PANC-1 cells. (E,G) Representative cell invasive images of elaiophylin-treated BxPC-3 and PANC-1 cells stained with crystal violet reagent. Magnification: 20×, scale bar: 50 µm. (F,H) The bar chart indicates the number of invaded cells in 24 h. (I) Representative western blotting images of MMP-7, MMP-2, and β-actin in BxPC-3 cells treated with elaiophylin at indicated concentrations. The original western blotting images are shown in Figure S5. (J,K) Bar charts indicate the ratio of MMP-7/β-actin and MMP-2/β-actin. Error bars, S.D.: * p < 0.05; ** p < 0.01; *** p < 0.001. Data are presented as mean ± SD from three independent experiments (n = 3).

Figure 6

Elaiophylin inhibits angiogenesis. (A) Representative images of elaiophylin-treated HUVECs showing the formation of capillary-like structures. Magnification: 10×, scale bar: 100 µm. (B) The bar chart indicates the number of junctions. (C) The bar chart indicates the number of nodes. Error bars, S.D.: *** p < 0.001. Data are presented as mean ± SD from three independent experiments (n = 3).

Figure 7

Elaiophylin inhibits the activation of the Wnt/β-Catenin signaling pathway in BxPC-3 cells. (A) Representative western blotting images of β-catenin, p-GSK-3β, LEF-1, and β-actin in BxPC-3 cells treated with elaiophylin at indicated concentrations. (B–D) Bar charts indicate the ratio of β-catenin/β-actin, p-GSK-3β/β-actin, and LEF-1/β-actin. The original western blotting images are shown in Figure S6. (E) The bar chart indicates the relative luciferase activity of the TOPFlash firefly luciferase. Error bars, S.D.: * p < 0.05; ** p < 0.01; *** p < 0.001. Data are presented as mean ± SD from three independent experiments (n = 3).