脂质体莪术醇逆转卵巢癌顺铂耐药作用的机制

Mechanism of liposome curcumol reversing cisplatin resistance in ovarian cancer

目的研究脂质体莪术醇(LC)对人卵巢癌细胞顺铂耐药的逆转作用及其机制。方法采用卵巢癌耐顺铂细胞株SKOV3/DDP,细胞分为5组:空白对照组、阴性对照组(含12.5μg/ml脂质体)、顺铂组(4μg/ml)、LC组(20μg/ml)、联合组[顺铂(4μg/ml)+LC(20μg/ml)],CCK-8检测各组细胞增殖能力,高效液相色谱-串联质谱法检测细胞内顺铂平均含量,流式细胞术检测细胞凋亡,qRT-PCR检测P糖蛋白(P-gp)的mRNA相对表达量,免疫细胞化学和Western blot检测各组细胞P-gp蛋白的表达情况。结果联合组细胞增殖能力最低,凋亡率最高(P<0.05);联合组细胞内顺铂平均含量大于顺铂组(P<0.05);联合组细胞的P-gp mRNA和蛋白相对表达量低于单用顺铂组(P<0.05)。结论LC可通过抑制P-gp的表达,降低人卵巢癌耐顺铂细胞株SKOV3/DDP对顺铂的耐药性。

Objective To investigate the effect and mechanism of liposome curcumol( LC) on cisplatin resistance of human ovarian cancer cells. Methods Ovarian cancer cisplatin resistant cell line SKOV3/DDP was used. Cells were divided into five groups: blank control group,negative control group( containing 12. 5 μg/ml liposome),cisplatin group( 4 μg/ml),LC group( 20 μg/ml),and combined group( cisplatin( 4 μg/ml) + LC( 20 μg/ml).Cell proliferation was detected by CCK8 assay,the content of cisplatin in cells was detected by high performance liquid chromatography-tandem mass spectrometry( HPLC-MS),and apoptosis was detected by flow cytometry. The level of P-glycoprotein( P-gp) mRNA was detected by qRT-PCR and the expression of P-gp protein was detected by immunohistochemistry and Western blot. Results The cell proliferation ability of the combination group was the lowest while the apoptosis rate was the highest( P < 0. 05) among the groups. HPLC-MS showed that the content of cisplatin in combined group was higher than that in cisplatin group( P < 0. 05). P-gp mRNA and protein in the combined group were down-regulated( P < 0. 05). Conclusion LC can reduce cisplatin resistance of human ovarian cancer cell line SKOV3/DDP by inhibiting the expression of P-gp.