Increasing knowledge of colorectal cancer stem cells (CCSCs) and tumor microenvironment improves our understanding of cellular mechanisms involved in the immunity against colorectal cancer (CRC). Tumor associated antigens were evaluated via RNA-seq and bioinformatics analysis, evoking promising targets for tumor immunotherapy. MUC1 has been demonstrated to participate in the maintenance, tumorigenicity, glycosylation and metastasis of CCSCs, which may provide a new priority for CSC vaccination. In the present study, the vaccination with CCSCs with high expression of MUC1 was evaluated in a murine model for the vaccine’s immunogenicity and protective efficacy against CRC. CD133⁺ CCSCs were isolated from SW620 cell line using a magnetic-activated cell sorting system, and shMUC1 was further used to knock down the expression of MUC1 in CD133⁺ CCSCs. Mice were subcutaneously immunized with the cell lysates of CCSCs and shMUC1 CCSCs, followed by a challenge with SW620 cells at ten days after final vaccination. The results indicated CCSC vaccine significantly reduced the tumor growth via a target killing of CCSCs as evidenced by a decrease of CD133⁺ cells and ALDH⁺ cells in tumors. Moreover, CCSC vaccine resulted in the elevated NK cytotoxicity, production of perforin, granzyme B, IFN-γ, memory B cells, and anti-MUC1 antibodies. Of note, MUC1 knockdown partly impaired the anti-tumor efficacy of CCSC vaccine. Importantly, the CCSC vaccine has no toxic damage to organs. Overall, CCSC vaccine could serve as a potent and safe vaccine for CRC treatment, and MUC1 might play an essential role in CCSC vaccine.

对结直肠癌干细胞（CCSC）和肿瘤微环境的了解的增加，增进了我们对参与抗结直肠癌（CRC）免疫力的细胞机制的了解。通过RNA-seq和生物信息学分析评估了肿瘤相关抗原，从而唤起了有希望的肿瘤免疫治疗靶标。已经证明MUC1参与CCSC的维持，致瘤性，糖基化和转移，这可能为CSC疫苗接种提供了新的优先事项。在本研究中，在鼠模型中评估了高表达MUC1的CCSC疫苗的免疫原性和对CRC的保护作用。CD133 ⁺使用磁激活细胞分选系统从SW620细胞系中分离出CCSC，并将shMUC1进一步敲低CD133 ⁺ CCSC中MUC1的表达。用CCSC和shMUC1 CCSC的细胞裂解物皮下免疫小鼠，然后在最终疫苗接种后第10天用SW620细胞攻击。结果表明，CCSC疫苗通过靶向杀死CCSCs显着降低了肿瘤的生长，CD133 ⁺细胞和ALDH ⁺的减少证明了这一点。肿瘤细胞。此外，CCSC疫苗导致NK细胞毒性升高，穿孔素，颗粒酶B，IFN-γ，记忆B细胞和抗MUC1抗体产生。值得注意的是，MUC1敲低部分削弱了CCSC疫苗的抗肿瘤功效。重要的是，CCSC疫苗对器官没有毒性损害。总体而言，CCSC疫苗可作为CRC治疗的有效且安全的疫苗，而MUC1可能在CCSC疫苗中发挥重要作用。