Effects of PHA-665752 and vemurafenib combination treatment on in vitro and murine xenograft growth of human colorectal cancer cells with BRAFV600E mutations

Zhi,Jie; Li,Zhongxin; Lv,Jian; Feng,Bo; Yang,Donghai; Xue,Liang; Zhao,Zhaolong; Zhang,Yanni; Wu,Jianhua; Jv,Yingchao; Jia,Yitao

doi:10.3892/ol.2018.7770

Effects of PHA-665752 and vemurafenib combination treatment on in vitro and murine xenograft growth of human colorectal cancer cells with BRAFV600E mutations

Authors:
- Jie Zhi
- Zhongxin Li
- Jian Lv
- Bo Feng
- Donghai Yang
- Liang Xue
- Zhaolong Zhao
- Yanni Zhang
- Jianhua Wu
- Yingchao Jv
- Yitao Jia
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Affiliations: Department of Oncology, Hebei General Hospital, Shijiazhuang, Hebei 050051, P.R. China, Second Department of Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China, Experimental Animal Center, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China
Published online on: January 10, 2018 https://doi.org/10.3892/ol.2018.7770
Pages: 3904-3910

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Abstract

It remains unknown whether blockade of B-Raf proto-oncogene, serine/threonine kinase (BRAF)V600E signaling and MET proto-oncogene, receptor tyrosine kinase (c‑Met) signaling is effective in suppressing the growth of human colorectal cancer (CRC) cells. The present study investigated the effects of the vemurafenib alone and in combination with c‑Met inhibitor PHA‑665752 on the growth of human CRC cells in vitro and in mouse xenografts. HT‑29 and RKO CRC cell lines with BRAFV600E mutations and mice bearing HT‑29 xenografts were treated with vemurafenib in the absence or presence of PHA‑665752. Cell viability and cycle phase were respectively examined by using the MTT and flow cytometry assay. Immunohistochemistry was conducted to detect the protein expression levels of hepatocyte growth factor (HGF), phosphorylated (p)-c‑Met, p‑AKT serine/threonine kinase (AKT) and p‑extracellular signal‑regulated kinase (p-ERK). The MTT assay demonstrated that the growth of RKO and HT‑29 cells was inhibited by PHA‑665752 in a time‑ and dose‑dependent manner (P<0.05), however no significant suppressive effects were observed with vemurafenib. Relative to the PHA‑665752 or vemurafenib stand‑alone treatment groups, the combination of PHA‑665752 and vemurafenib had a significant inhibitory effect on the proliferation of CRC cell lines (P<0.05). The mean tumor volume in mice treated with vemurafenib in combination with PHA‑665752 was significantly smaller compared with those treated with only vemurafenib or PHA‑665752 (P<0.05). Flow cytometry assay revealed that the G0/G1 phase frequency was significantly increased in the combination group compared with any other treatment groups (P<0.05). Immunohistochemistry demonstrated that vemurafenib in combination with PHA‑665752 effectively induced the expression of p‑c‑Met, p‑AKT and p‑ERK, however had no effect on HGF.

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