Weichang’an Formula Inhibits Tumor Growth in Combination with Bevacizumab in a Murine Model of Colon Cancer—Making up for the Deficiency of Bevacizumab by inhibiting VEGFR-1

Abstract

Aim: Angiogenesis plays an important role in the initiation, development, and metastasis of malignant tumors. Antiangiogenic drugs combined with immune therapy are considered to have a synergistic effect on anti-tumor strategy. Weichang’an formula (WCAF) is a prescription of traditional Chinese medicine (TCM) based on pharmaceutical screening and clinical experience. The aim of this study is to examine the effect of WCAF and its combined action with Bevacizumab (BEV) in colorectal cancer, and to identify the possible mechanism of action.Methods: A human colon cancer cell (HCT 116) subcutaneous xenograft model was established in BALB/c-nu/nu mice. Tumor-bearing mice were randomized into each of four groups: control, WCAF treated, BEV treated, and WCAF plus BEV treated. Apoptosis was detected by TUNEL assay. Western blot was used to assess the protein levels of Leptin-R, STAT3, p-STAT3, BCL-2, and VEGFR-1. Immunohistochemistry was used to detect the micro-vessel density (MVD) and AKT1. Leptin and Vascular endothelial growth factor A (VEGF-A) mRNA expression were detected by Real-time PCR (RT-PCR). A network pharmacology study and validation assay were carried out to find the underlying molecular targets of WCAF related to immune regulation.Results: Compared with the control group, WCAF reduced tumor weight and volume, as well as promoted tumor cell apoptosis. WCAF treatment decreased the mRNA expression of Leptin and VEGF-A, while the protein levels of CD31, LEP-R, VEGFR-1, STAT3, and p-STAT3 were decreased in tumor tissues. In addition, VEGFR-1 protein expression was decreased in the WCAF group and the WCAF plus BEV group but not in the BEV group. The combination of WCAF and BEV demonstrated a partial additive anti-tumor effect in vivo. The pharmacological network also found there are 26 WCAF target proteins related to cancer immune and 12 cancer immune related pathways. The AKT1 protein expression in the WCAF and WCAF + BEV groups were significantly lower than the that in the control group (p < 0.01).Conclusion: WCAF can inhibit tumor growth and promote apoptosis and inhibit tumor angiogenesis in subcutaneous xenografts of human colon cancer HCT-116 in nude mice. WCAF also makes up for the deficiency of BEV by inhibiting VEGFR-1. The VEGFR-1 expression between the combination group and BEV alone achieved statistically significant difference (p < 0.01). Combined with BEV, WCAF showed a partial additive anti-tumor effect. The mechanism may be related to Leptin/STAT3 signal transduction, VEGF-A, VEGFR-1 and WCAF target proteins related to cancer immune such as leptin and AKT1.

Erlotinib plus bevacizumab versus erlotinib alone in patients withEGFR-positive advanced non-small-cell lung cancer: a systematic review and meta-analysis of randomised controlled trials

“…Another possible mechanism is that bevacizumab may restore cell apoptosis by inhibiting the VEGF-mediated pathway. 34 Due to synergistic inhibition of cancer growth signalling, VEGF signal inhibition is still effective for cancers with EGFR TKI resistant mutations. 35 An animal study 36 suggested that erlotinib plus bevacizumab treatment restored resistance to the VEGF-mediated pathway.…”

“…Therefore, a higher intratumoural concentration of erlotinib could prolong resistance to TKIs. Another possible mechanism is that bevacizumab may restore cell apoptosis by inhibiting the VEGF-mediated pathway 34. Due to synergistic inhibition of cancer growth signalling, VEGF signal inhibition is still effective for cancers with EGFR TKI resistant mutations 35.…”

Alizarin, a nature compound, inhibits the growth of pancreatic cancer cells by abrogating NF-κB activation

“…Recently, an increasing number of researchers have begun to pay attention to nature compounds which have good antitumor capability and almost no obvious side effects 26 . A growing body of studies has shown that both Chinese herbs and their active components can directly inhibit tumors by inhibiting angiogenesis 27 , blocking the tumor cell growth cycle 28 - 30 , or inducing tumor cell apoptosis 31 , 32 and indirectly inhibit tumors by regulating the tumor microenvironment; for example, by increasing the activity of NK cells 33 , downregulating the immunosuppressive activity of myeloid-derived suppressor cells 28 , 34 , and improving the immune response of dendritic cells 35 . Our lab has been committed to revealing the mechanism of the antitumor activity of nature compounds for years 28 , 30 , 31 , 33 .…”

Ginsenoside compound K inhibits the proliferation, migration and invasion of Eca109 cell via VEGF-A/Pi3k/Akt pathway

“…Furthermore, the therapeutics values of Ginsenoside CK in tumors have well been studied in bladder cancer, colon cancer, liver cancer which can significantly suppress the proliferation ability of these tumor cells [ 7 – 9 ]. Vascular endothelial growth factor-A (VEGF-A), a highly specific pro-vascular endothelial cell growth factor, which can promote extracellular matrix degeneration, vascular permeability, proliferation, migration and angiogenesis of vascular endothelial cells [ 10 , 11 ]. Study suggested that Ginsenoside CK inhibit tumor angiogenesis by suppressing the proliferation and migration of vascular endothelial cells, inhibiting the activity of VEGF-A and it’s signaling pathway, and the degradation of vascular extracellular matrix in neuroblastoma cells [ 12 ].…”