Volume 29, Issue 5, 2020
DOI: 10.24205/03276716.2020.1106
Cetuximab Combined with Vemurafenib Can Significantly Inhibit the Proli- Feration Of Braf V600e Mutant Egfr Wild-Type Colorectal Cancer Cells
Abstract
Background: Cetuximab (Cetuximab, C225) is relatively limited in improving the survival of patients with advanced colorectal cancer. One possible reason is that some patients with colorectal cancer have high-frequency mutations of BRAF V600E leading to resistance. Vemurafenib is a newly developed inhibitor that specifically blocks the activation of signaling pathways caused by BRAF V600E mutations. Therefore, this study thoroughly studied whether the combined application of C225 and Vemurafenib can inhibit the proliferation of EGFR wild-type, BRAF V600E mutant colorectal cancer cells and the molecular mechanism.
Methods: The effects of different concentrations of C225 and BRAF V600E potent mutant (Vemurafenib) on cell proliferation of colorectal cancer cell lines HT-29 (EGFR wild-type and BRAF V600E mutant) were determined by using CCK-8 and plate cloning assay.Western Blot was used to detect the changes in apoptosis (Caspase-3) and proliferation-related markers (Ki-67, CD31) protein levels of cells treated with C225 alone, Vemurafenib alone or in combination with the two drugs.
Results: The results of CCK-8 clone formation experiment showed that the combined application of C225 and Vemurafenib can significantly reduce cell viability and cell clone formation ability compared with the two single-agent groups. After treatment of cells with the combination of C225 and Vemurafenib, the activity of Caspase-3 was increased, the expression of CD31 and Ki67 was significantly inhibited, the expression of p-EGFR was down-regulated, the total amount of ERK decreased, and the corresponding phosphorylated p-ERK level appeared Elevated.
Conclusion: The combined application of C225 and Vemurafenib can significantly inhibit the proliferation and clone formation of colorectal cancer cell lines. When used in combination, the expression of Caspase-3 was significantly increased, and the expression of Ki-67, CD31 and EGFR phosphor - lation was inhibited. This study provides a theoretical basis for the combined administration of EGFR monoclonal antibody C225 and Vemurafenib for clinically EGFR wild-type and BRAF V600E mutant colon cancer patients, provides preliminary theoretical basis and support for subsequent clinical research.
Keywords
Cancer, Cetuximab, Vemurafenib, BRAF V600E Mutant, Proliferation