沉默泡膜蛋白1表达提高结肠癌细胞对5-氟尿嘧啶的敏感性

Silencing vacuole membrane protein 1 increases colon cancer cell sensitivity to 5-fluorouracil

目的分析沉默泡膜蛋白1(vacuole membrane protein 1,VMP1)与结肠癌细胞对药物敏感性的关系及其机制。方法应用MTT法检测结肠癌细胞的药物敏感性;构建沉默VMP1慢病毒载体,感染SW620细胞沉默其VMP1表达;软琼脂克隆形成实验检测沉默VMP1后细胞在5-氟尿嘧啶作用下的单克隆形成及增殖能力;Western blot检测耐药基因ABCC1蛋白表达水平。结果SW620细胞对5-氟尿嘧啶的敏感性要显著低于铂类;沉默VMP1后,SW620细胞对5-氟尿嘧啶的敏感性显著增加,5-氟尿嘧啶作用下的单细胞克隆形成能力被显著抑制,ABCC1表达水平显著降低。结论抑制VMP1表达可能通过降低耐药基因ABCC1的表达提高其对5-氟尿嘧啶的敏感性。

Objective To analyze the relationship between vacuole membrane protein 1(VMP1)expression and the drug sensitivity of colon cancer cells and its mechanism.Methods The drug sensitivity of colon cancer cells was evaluated by MTT.SW620 cells were infected with a lentiviral vector to silence VMP1 expression.Soft agar clone formation assay was used to detect the monoclonal formation and proliferation of VMP1 silenced cells in the presence of 5-fluorouracil.Western blot was used to detect the protein expression of drug resistance gene ABCC1.Results The sensitivity of SW620 cells to 5-fluorouracil was significantly lower than that to platinum.After silencing VMP1,the sensitivity of SW620 cells to 5-fluorouracil was significantly increased.5-fluorouracil inhibited the ability of single-cell clone formation and significantly reduced the expression of ABCC1.Conclusion Inhibition of VMP1 expression may increase the sensitivity of colon cancer cells to 5-fluorouracil by down-regulating drug resistance gene ABCC1.