二十二碳六烯酸对人胆管癌细胞增殖和凋亡的影响

Effect of docosahexaenoic acid on cell growth and apoptosis of human cholangiocarcinoma cell

目的研究二十二碳六烯酸(DHA)对人胆管癌QBC 939细胞增殖、凋亡和周期的影响,并初步探讨可能的分子机制。方法采用MTT法检测DHA和花生四烯酸(AA)对QBC 939细胞增殖的影响;流式细胞术检测DHA对QBC 939细胞凋亡和周期的影响;免疫印迹法检测DHA作用于QBC 939细胞后β-catenin、c-myc和cyclin D1三种蛋白表达量的变化。结果作用浓度为80和100μg/mL的DHA可显著降低QBC 939细胞存活率,且呈一定的剂量、时间效应(P<0.05),AA无明显作用。经80μg/mL DHA作用24 h后,QBC 939细胞早期凋亡率明显增加(P<0.05),G0/G1期细胞显著增多(P<0.05),G2/M、S期细胞减少(分别P<0.01及P<0.05)。β-catenin、c-myc、cyclin D1表达量均下调。结论 DHA可抑制人胆管癌QBC939细胞增殖,促进其凋亡,部分作用机制可能是下调Wnt/β-catenin信号传导通路中β-catenin、c-myc和cyclin D1三种蛋白的表达。

【Objective】 To assess the effects of Docosahexaenoic Acid（DHA） on cell viability,apoptosis and cell cycle of human cholangiocarcinoma cell line QBC 939 in vitro and to study its possible mechanism.【Methods】 Cell cycle and apoptosis of the cells were determined by MTT assay and flow cytometry;the expression of β-catenin,c-myc,cyclin D1 was determined by western blot.【Results】 After treatment with 80～100 μg/mL of DHA,the cell viability was significantly decreased（P 〈0.05） while represented no marked change with same dosage of arachidonic acid（AA）.Apoptosis on the treated cells was increased（P 〈0.05） and cells at G2/M phase and S phase were decreased significantly（P 〈0.01,P 〈0.05 respectively）,whereas G0/G1 phase was increased（P 〈0.05）.The expression of β-catenin,c-myc and cyclin D1 significantly decreased compared with control group.【Conclusions】 DHA can inhibit the growth of human cholangiocarcinoma QBC 939 cells and the possible molecular mechanism may be involved partly in downgrading of Wnt/β-catenin pathway.