Fig. 4: HKDC1 promotes proliferation, migration, invasion, and immune escape of OC cells through lipid metabolism in vitro.

A HDAC1 mRNA and protein expression levels in SKOV3/ID8 cells with HKDC1 knockdown and treated with 150 μM FFA were evaluated using qRT-PCR and WB analysis. B Intracellular metabolite levels, including FFA, TG, PL, and CHO, were quantified in SKOV3/ID8 cells with HKDC1 knockdown and treated with 150 μM FFA using biochemical kits. C Neutral lipid accumulation was assessed via BODIPY 493/503 staining in SKOV3/ID8 cells with HKDC1 knockdown and treated with 150 μM FFA. D The mRNA and protein expression levels of ACC1, FASN, SCD1, HMGCS1, HMGCR, CD36, and CPT1A were determined by qRT-PCR and WB in SKOV3/ID8 cells with HKDC1 knockdown and treated with 150 μM FFA. E Cell growth was evaluated through the CCK-8 proliferation assay in SKOV3/ID8 cells with HKDC1 knockdown and treated with 150 μM FFA. F Colony formation assays assessed the proliferative capacity of SKOV3/ID8 cells under the same conditions. G Wound healing assays were performed at 0 h, 24 h, and 48 h to examine the migration potential of SKOV3/ID8 cells with HKDC1 knockdown and treated with 150 μM FFA. H Transwell invasion assays were used to determine the invasive potential of SKOV3/ID8 cells with HKDC1 knockdown and treated with 150 μM FFA. I WB analysis was performed to assess PD-L1 protein expression in SKOV3/ID8 cells with HKDC1 knockdown and treated with 150 μM FFA. J Flow cytometry was employed to investigate the proportions of T cells (CD45⁺CD4⁺/CD8⁺), Treg cells (CD4⁺CD25⁺FOXP3⁺), and NK cells (CD3⁻CD16⁺CD56⁺) within PBMCs co-cultured with SKOV3/ID8 cells with HKDC1 knockdown and treated with 150 μM FFA. K Flow cytometry analysis assessed PD-1 expression on T cells and NK cells, as well as IFN-γ production within CD4⁺/CD8⁺ T cells in PBMCs co-cultured with HKDC1-modulated SKOV3/ID8 cells treated with 150 μM FFA. L qRT-PCR was used to analyze PD-1 mRNA levels in T cells and NK cells from PBMCs co-cultured with SKOV3/ID8 cells with HKDC1 knockdown and treated with 150 μM FFA. M ELISA was performed to measure GZMB and perforin levels in NK cells from PBMCs co-cultured with HKDC1-knockdown SKOV3/ID8 cells treated with 150 μM FFA. ns not significant, ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, ^****P < 0.0001, one-way ANOVA test. n = 3 biologically independent samples. Data are expressed as mean ± standard deviation.