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Archive > Volume 23 Issue 6 > 2016,23(6):751-758. DOI:10.3872/j.issn.1007-385X.2016.06.003 Prev Next
Prompt Report
Killing effect of DC-CIK/CTL loaded on membrane microparticles of the cancer stem cell synergisted with cetuximab on colorectal cancer cells and the mechanism of its action
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Abstract:
Objective: To explore killing effect of DC-CIK/CTL loaded on membrane microparticles (MMPs) of the colorectal cancer stem cells synergisted with cetuximab (C225) on colorectal cancer cells which are resistent to epidermal growth factor receptor (EGFR)-targeting drug C225, and their mechanisms. Methods: Mutation satus of k-RAS gene for colorectal cancer cell SW480, SW620 and HCT116 line cells were identified by PCR. Cancer stem cells of the SW480 and HCT116 were enriched with serum free suspension cell culture method. Stem cell markers, Sox-2 and Oct-4, were detected by RT-PCR. Pheripheral blood monocuclear cells were extracted to culture DC and CIK, CIK was co-cultured with DC loaded with MMP of the colorectal cancer stem cells. Observation of mophology and MTT assay were respectively used to detect killing effects of DC-CIK/CTL cells, their culture supernatant (DC-CIK/CTL’S) and C225 alone as well as both combination on the SW480, SW620, HCT116 line cells and their stem cells. Exprissions of apoptosis-related gene Fas, epithelial-mesenchymal transition (EMT)-related proteins, E-cadherin and vinmentin, in the SW480, SW620, HCT116 line cells after treatmented by DC-CIK/CTL’S and C225 alone as well as both combination were detected by RT-PCR. Effects of DC-CIK/CTL’S and C225 alone as well as both combination on invasion acts of the colorectal cance cells were detected by a scratch assay. Results: All of the SW480, SW620 and HCT116 cell lines were identified as K-RAS mutation type. Stem cells of the colorectal carcinoma which obtained from enrichment culture had a high expression of the stem cell marker Sox-2 and Oct-4. The DC-CIK/CTL cells loaded with MMPs, thier secretion supernatant and C225 had a synergistic effect on inhibition of colorectal cancer cells. the combination group of C225 with DC-CIK/CTL’S enhanced expressions of Fas and E-cadherin proteins, comparing with groups of C225 and DC-CIK/CTL’S alone. Migration rate of the cells in the combination treatment group was smaller than those in the single treatment groups. Conclusion: The DC-CIK/CTL loaded with the MMPs of colorectal cancer stem cell could be of specific targeting killing effect in vitro on the colorectal cancer cells which are resistant to EGFR-target drug. It could have a obvious synergistic effect with C225. Their mechanisms could be related to reverse of EMT and improvement of apoptosis of the cells.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 31170880, 81371891)
Clc Number:
