Qin Xiaoning, Ruan Hongxun, Yuan Liqing, Lin Lin
The Third Department of General Surgery, The Second Hospital of Hebei Medical University, Shijiazhuang, 050000 Hebei China.
The Second Department of Gynaecology, The Second Hospital of Hebei Medical University, Shijiazhuang, 050000 Hebei China.
3 Biotech. 2023 Oct;13(10):327. doi: 10.1007/s13205-023-03742-5. Epub 2023 Sep 1.
Bevacizumab is the standard treatment for colorectal cancer (CRC) in the advanced stage. However, poor diagnosis identified due to the bevacizumab resistance in many CRC patients. Previous studies have found that CRC stem cells (CCSCs) and interleukin 22 (IL-22) are involved in the resistance of bevacizumab, however, the mechanism of remains unclear. In this study, we established the bevacizumab drug-resistant cell line HCT-116-R by concentration gradient method, and the cell viability was detected by CCK-8 assay. The resistance of bevacizumab in CRC cell lines HCT-116-R was identified by characterizing epithelial-mesenchymal transition (EMT). Additionally, HCT-116-R cell lines were isolated from CCSCs and their tumorigenicity was validated in nude mice. We observed that that compared with the matched group, the expression of IL-22, IL-22R, STAT3, and GP130 in drug-resistant cells increased distinctly, with blocked IL-22 cells were successfully constructed by lentiviral interference. The level of proteins in stem cell landmarks (EpCAM, CD133), and stem cell landmarks (Oct4, Sox2) was identified by western blotting. Furthermore, the IL-22 role was evaluated by xenograft model. We found that short hairpin RNA (shRNA) suppression of IL-22 expression can restore the sensitivity of drug-resistant CCSCs to bevacizumab, Moreover, xenograft tumor models show that suppression of IL-22 can increase the anti-tumor influence of bevacizumab. In summary, we demonstrated that CCSCs play a major part in bevacizumab-resistant CRC. Inhibiting the signaling pathway of IL-22/STAT3 can improve the anti-tumor influence on bevacizumab in vitro and in vivo. Thus, IL-22 may represent a new anti-bevacizumab target in CRC.
贝伐单抗是晚期结直肠癌(CRC)的标准治疗药物。然而,许多CRC患者因贝伐单抗耐药而导致诊断不佳。先前的研究发现,CRC干细胞(CCSCs)和白细胞介素22(IL-22)与贝伐单抗耐药有关,但其机制仍不清楚。在本研究中,我们通过浓度梯度法建立了贝伐单抗耐药细胞系HCT-116-R,并通过CCK-8法检测细胞活力。通过上皮-间质转化(EMT)特征鉴定了CRC细胞系HCT-116-R对贝伐单抗的耐药性。此外,从CCSCs中分离出HCT-116-R细胞系,并在裸鼠中验证了其致瘤性。我们观察到,与匹配组相比,耐药细胞中IL-22、IL-22R、STAT3和GP130的表达明显增加,通过慢病毒干扰成功构建了IL-22阻断细胞。通过蛋白质免疫印迹法鉴定干细胞标志物(EpCAM、CD133)和干细胞标志物(Oct4、Sox2)中的蛋白质水平。此外,通过异种移植模型评估了IL-22的作用。我们发现,短发夹RNA(shRNA)抑制IL-22表达可恢复耐药CCSCs对贝伐单抗的敏感性,此外,异种移植肿瘤模型表明,抑制IL-22可增强贝伐单抗的抗肿瘤作用。总之,我们证明了CCSCs在贝伐单抗耐药的CRC中起主要作用。抑制IL-22/STAT3信号通路可在体外和体内增强贝伐单抗的抗肿瘤作用。因此,IL-22可能是CRC中一种新的抗贝伐单抗靶点。