摘要
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 explore the inhibition effects of bafilomycin A1 on the autophagy of ovarian cancer cells SKOV3 handled by 5-fluorouracil （5-FU） and the possible mechanism of enhancing chemotherapy sensitivity. Methods： SKOV3 cells were divided into 3 groups, group A were treated with 5-FU （50 pmol/L） for 48 h, group B were treated with 100 nmolJL bafilomycin A1 for 1 h, then treated for 48 h with 50 pmol/L 5-FU, group C were cultured without any treatment as the negative control group. The morphologic changes of ovarian cancer cells SKOV3 with autophgay were observed by acridine orange staining and indirect immunofluotesent assay. The expression of LC3 and Beclinl proteins in SKOV3 cells were also detected by Western blotting. Results： Acidine orange staining showed that the acid areas were markedly decreased in group B. And the immunofluotesent assay showed the treatment of bafilomycin A1 weakened the formation of autophagy. Furthermore, the expression levels of LC3 and Beclinl proteins in SKOV3 cells were significantly reduced after the treatment of bafilomycin A1. Conclusion： Bafilomycin A1 might inhibit the autophagy of SKOV3 cells by reducing the number of acid areas in SKOV3 cells and inhibiting the expression of LC3 and Beclinl proteins in SKOV3 cells.
出处
《生殖与避孕》
CAS
CSCD
2013年第6期367-371,共5页
Reproduction and Contraception
