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7A). Next, we used PDX models and HCT15 cell-derived xenografts to assess the combination of ML347 and Regorafenib in vivo. When tumours were palpable, the mice were randomly divided into four groups and treated respectively with vehicle, ML347, Regorafenib, Regorafenib and ML347 in combination. Regorafenib alone could reduce tumour growth, ML347 alone showed weak tumour inhibition ability. However, the combination of Regorafenib and ML347 completely inhibited tumour growth in vivo (Fig. 7B, E and Additional file 1: Figure S5A). Tumour cell apoptosis was further assessed by TUNEL assay in sections of PDX models and HCT15 cell-derived xenografts. There were significantly more apoptotic cells in the combined treatment group than in the ML347 or Regorafenib treatment groups (Fig. 7D, F and Additional file 1: Figure S5B). By ISH and IHC staining, we observed a same pattern for the expressions of ACVRL1 and GPX2 under different drug treatment, but for miR-7-5p, no obvious change was seen in different drug treatment groups (Fig. 7C). Taken together, these data confirm that ACVRL1 inhibition could effectively sensitize colon cancer cells to Regorafenib treatment in vivo Fig. 8.Fig. 7ACVRL1 inhibition sensitizes CRC to mTKI treatment in vitro and in vivo. A Annexin V-PI staining showed that Regorafenib can induce apoptosis of HCT15 cells, and Regorafenib combined with ML347 significantly increased the apoptosis ratio of HCT15 cells. B, E Regorafenib inhibited CRC tumourigenicity. ML347 and Regorafenib combination therapy effectively abolished CRC growth in PDX#1 model and nude mice model. C The representative tumour tissue sections from PDX#1 model in different drug treatment groups were analysed by IHC and ISH for the expressions of miR-7-5p, ACVRL1 and GPX2. Scale bar, 30 μm. D, F TUNEL staining of the transplanted tumour tissues of PDX#1 and nude mice models showed that Regorafenib combined with ML347 increased the ratio of tumour

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