Department of Pharmacy, The First People's Hospital of Taicang, Taicang, PR China; The Affiliated Taicang Hospital of Suzhou University, Taicang, PR China.
Department of Pharmacy, The First People's Hospital of Taicang, Taicang, PR China; The Affiliated Taicang Hospital of Suzhou University, Taicang, PR China.
Tissue Cell. 2022 Aug;77:101851. doi: 10.1016/j.tice.2022.101851. Epub 2022 Jun 7.
Drug resistance of colon cancer cells is the key to affect the efficacy of colon cancer chemotherapy and lead to chemotherapy failure. Recent studies have found that exosomes play an important role in chemoresistance of colon cancer, while the expression of VMP1 may be involved in exosome secretion.
Drug sensitivity of colon cancer cells was detected by MTT. VMP1 expression levels were detected by qPCR and western blot. Expression of VMP1 was silenced in SW620 and HT-29 cell lines. Exosomes were isolated by ultracentrifugation, the particle size and concentration of exosomes were analyzed by NTA, and the morphology of exosomes was investigated by transmission electron microscopy. Exosome marker protein expression was detected by Western blot. The effect of exosomes on 5-FU sensitivity in SW620 and HT-29 cells was examined by MTT and western blot.
VMP1 presented high expression in SW620 and HT-29 cells, their VMP1 gene and protein levels were significantly higher than those in SW480 and HCT116 cells, they were less sensitive to 5-FU, and exosome secretion was significantly reduced in SW620 and HT-29 cells after silencing of VMP1. Importantly, addition of exosomes to cells after silencing of VMP1 re-improved drug resistance. Co-incubation of exosomes secreted from SW620 and HT-29 cells with homologous cells significantly increased cell viability after 5-FU (50 μM) treatment, and increased the expression levels of the resistance protein ABCC1 and the anti-apoptotic protein Bcl-2.
High expression of VMP1 in colon cancer cells is able to promote exosome secretion, which mediates the acquisition of more drug-resistant properties by cancer cells.
结肠癌耐药性是影响结肠癌化疗疗效、导致化疗失败的关键。最近的研究发现，外泌体在结肠癌耐药中发挥重要作用，而 VMP1 的表达可能参与外泌体的分泌。
MTT 法检测结肠癌细胞的药物敏感性。qPCR 和 Western blot 检测 VMP1 的表达水平。沉默 SW620 和 HT-29 细胞系中的 VMP1 表达。超速离心分离外泌体，NTA 分析外泌体的粒径和浓度，透射电镜观察外泌体的形态。Western blot 检测外泌体标志蛋白的表达。MTT 和 Western blot 检测外泌体对 SW620 和 HT-29 细胞 5-FU 敏感性的影响。
VMP1 在 SW620 和 HT-29 细胞中高表达，其 VMP1 基因和蛋白水平明显高于 SW480 和 HCT116 细胞，对 5-FU 的敏感性较低，沉默 VMP1 后 SW620 和 HT-29 细胞的外泌体分泌明显减少。重要的是，沉默 VMP1 后加入外泌体可重新提高细胞的耐药性。共孵育 SW620 和 HT-29 细胞分泌的外泌体后，5-FU（50μM）处理后细胞活力明显增加，耐药蛋白 ABCC1 和抗凋亡蛋白 Bcl-2 的表达水平也增加。
结肠癌细胞中 VMP1 的高表达能够促进外泌体的分泌，从而介导癌细胞获得更多的耐药特性。
