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目的:研究巴弗洛霉素A1(bafilomycin A1)对5-氟尿嘧啶(5-FU)作用下的卵巢癌细胞SKOV3自噬的抑制作用及其增强化疗敏感性的可能机制。方法:将SKOV3细胞随机分为3组,A组:50μmol/L5-氟尿嘧啶(5-FU)处理48h;B组:100nmol/L巴弗洛霉素A1给药1h后加入50μmol/L5-FU处理48 h;C组(阴性对照):不作任何处理。3组细胞均应用吖啶橙染色、间接免疫荧光技术检测卵巢癌细胞SKOV3自噬形态学变化;并通过Western blotting检测巴弗洛霉素A1对SKOV3细胞中微管相关蛋白轻链3(LC3)及自噬相关蛋白Beclin1表达的影响。结果:B组产生的红色酸性区域明显减少,LC3定位的荧光亮度减弱,细胞内LC3、Beclin1蛋白表达量显著降低。结论:巴弗洛霉素A1可能通过减少SKOV3细胞酸性区域的数量,抑制LC3产生从而抑制SKOV3细胞的自噬。
OBJECTIVE: To study the inhibitory effect of bafilomycin A1 on the autophagy of ovarian cancer cell line SKOV3 induced by 5-fluorouracil (5-FU) and its possible mechanism of enhancing chemosensitivity. Methods: The SKOV3 cells were randomly divided into three groups: group A: 50μmol / L 5-fluorouracil (5-FU) for 48h; group B: 100μmol / L bafilomycin A1 48 h; Group C (negative control): without any treatment. The acridine orange staining and indirect immunofluorescence assay were used to detect the morphological changes of autophagy in ovarian cancer cell line SKOV3. Western blotting was used to detect the expression of microtubule-associated protein 3 (LC3) And autophagy-related protein Beclin1 expression. Results: In group B, the red acidic region was significantly reduced, the fluorescence intensity of LC3 localization was weakened, and the expression of LC3 and Beclin1 in cells was significantly decreased. CONCLUSION: Bafilomycin A1 inhibits the autophagy of SKOV3 cells by decreasing the number of acidic regions of SKOV3 cells and inhibiting the production of LC3.