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目的观察脂多糖诱导的SKOV3/DDP细胞内分泌型一氧化氮合酶(iNOS)表达变化对人卵巢癌SKOV3/DDP细胞株顺铂耐药性的影响,并初步探讨机制。方法体外培养SKOV3/DDP细胞,分为对照组、顺铂组、脂多糖(Lipopolysaccharides,LPS)组和联合用药组共4组,对照组给予常规不加血清培养基,顺铂组给予10μg/ml顺铂,LPS组给予浓度为1μg/ml的LPS,联合用药组给予10μg/ml顺铂和1μg/ml LPS,均作用24 h。MTT法检测各组细胞活力;蛋白免疫印迹法观察各组iNOS表达;流式细胞术检测各组细胞凋亡情况;间接免疫荧光法观察各组细胞激活型Caspase-3表达。结果联合用药组细胞活力明显低于顺铂组(P<0.05);顺铂组与对照组iNOS表达差异无统计学意义(P>0.05),LPS组和联合用药组iNOS表达水平明显高于对照组(P<0.05),且联合用药组高于LPS组(P<0.05);流式细胞术结果显示联合用药组细胞凋亡率明显高于顺铂组(P<0.05),且通过间接免疫荧光法显示联合用药组细胞激活型Caspase-3表达高于顺铂组(P<0.05)。结论 LPS能够降低人卵巢癌SKOV3/DDP细胞顺铂耐药性,这可能与其提高SKOV3/DDP细胞内iNOS表达水平有关。
Objective To investigate the effects of lipopolysaccharide on the expression of iNOS in SKOV3 / DDP cells induced by lipopolysaccharide (LPS) on cisplatin-resistant human ovarian cancer cell line SKOV3 / DDP and its possible mechanism. Methods SKOV3 / DDP cells were cultured in vitro and divided into 4 groups: control group, cisplatin group, lipopolysaccharide (LPS) group and combination group. The control group was given normal serum-free medium, while the cisplatin group was given 10μg / ml Cisplatin and LPS groups were given LPS at a concentration of 1μg / ml, and the combination group was treated with 10μg / ml cisplatin and 1μg / ml LPS for 24 hours. Cell viability was measured by MTT assay. INOS expression was observed by Western blotting. Apoptosis was detected by flow cytometry. Caspase-3 expression was detected by indirect immunofluorescence. Results The cell viability of the combination group was significantly lower than that of cisplatin group (P <0.05). The expression of iNOS in cisplatin group and control group was not significantly different (P> 0.05). The expression of iNOS in LPS group and combination group was significantly higher than that in control group (P <0.05), and the combined treatment group was higher than that of LPS group (P <0.05). The results of flow cytometry showed that the apoptosis rate of the combination group was significantly higher than that of cisplatin group (P <0.05), and by indirect immunofluorescence The results showed that the expression of cell-activated Caspase-3 in combination group was higher than that in cisplatin group (P <0.05). Conclusion LPS can reduce the cisplatin resistance of human ovarian cancer SKOV3 / DDP cells, which may be related to the increase of iNOS expression in SKOV3 / DDP cells.