Fig. 2: Knockdown of ABCA8 in Gem-R PC cells improves GEM sensitivity in vitro and in vivo. | Cell Death Discovery

Fig. 2: Knockdown of ABCA8 in Gem-R PC cells improves GEM sensitivity in vitro and in vivo.

From: ABCA8-mediated efflux of taurocholic acid contributes to gemcitabine insensitivity in human pancreatic cancer via the S1PR2-ERK pathway

Fig. 2

ABCA8 was stably knocked down in PANC-1 or CFPAC-1 Gem-R cells using shRNA targeting the ABCA8 gene (shABCA8). A scrambled shRNA (shScr) was used as a control. A Western blot analyses of ABCA8, BAX, and BCL2 protein expression in shScr or shABCA8 Gem-R cells. The histogram shows the densitometric analysis of the bands (n = 3 independent biological repeats). B GEM sensitivity of Gem-R shABCA8 cells compared with that of shScr cells was determined by CCK-8 cytotoxicity assays (n = 4 independent biological repeats). Histograms show the comparison of IC50 values between Gem-R shABCA8 cells and shScr cells. C–F BALB/c nude mice were subcutaneously injected with PANC-1 Gem-R shScr or shABCA8-1 cells and then treated with GEM (50 mg/kg, 4 times/week) or PBS as a control (n = 8 for each group). Growth curves of Gem-R shScr tumors and shABCA8 tumors were shown in C. The subcutaneous tumors were collected and weighed at the time of killing (3 weeks after treatment). Three representative tumors for each group are shown in D. E Comparison of tumor weight upon necropsy between groups. F Comparison of the tumor inhibition rate of GEM therapy between PANC-1 Gem-R shScr and shABCA8 tumors. G shScr- or shABCA8-transfected Gem-R PANC-1 or CFPAC-1 cells were treated with 1 or 5 μM GEM for 72 h and cell apoptosis was detected by annexin V/7-AAD staining followed by flow cytometry (n = 4 independent biological repeats). *P < 0.05, **P < 0.01, and ***P < 0.001.

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