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. 2021 Dec 31;33(6):694–707. doi: 10.21147/j.issn.1000-9604.2021.06.06

Figure 4.

Figure 4

ATRA combination therapy suppresses growth of recurrent HCC after IRFA by inhibition of TICs. (A) Representative phase contrast micrographs of the spheroid forming efficiency of HepG2 cells treated with or without heat and ATRA; (B) Tumor growth of different administered groups of HepG2 xenografts in NOD/SCID mice (n=3/group). Tumor size was measured with a caliper rule every 2 days; (C) Photographs show tumors and tumor weight for each group of HepG2 xenografts; (D) Kaplan-Meier survival curves of NOD/SCID mice after the treatments showed the combination therapy of RFA and ATRA (red line) had the best survival outcome; (E−G) Combination therapy of 20 mg/kg ATRA indicated slowest tumor growth (E), smallest tumor weight (F) and longest survival time (G) in the SK-Hep-1 xenografts (n=3/group); (H) Immunofluorescence staining of CD133+ and EpCAM+ cells in HepG2 xenograft. Positive cells were stained as red signal; nuclei were stained with DAPI (blue); (I) Proportions of CD133+ and EpCAM+ subpopulation of HepG2 xenografts with different treatment were analyzed by FCA; (J) Expression of the stem cell-related genes was inhibited in the combination treatment tumors compared with the RFA alone by Western blot analysis; (K−M) Tumor diameter (measured with a caliper rule every 2 days), photographs and tumor weight of the second HepG2 xenografts treated with 20 mg/kg ATRA or combination therapy. Data in (A, I) are presented as Inline graphic . ATRA, all-trans retinoic acid; HCC, hepatocellular carcinoma; IRFA, insufficient radiofrequency ablation; TIC, tumor-initiating cell; RFA, radiofrequency ablation; EpCAM, epithelial cell adhesion molecule; DAPI, 4’,6-diamidino-2-phenylindole; FCA, flow cytometric analysis. *, P<0.05;**, P<0.01;***, P<0.001.