廿烷五烯酸协同顺铂对肺腺癌细胞株的影响

Synergistic effect of eicosapentaenoic acid and cisplatin on A-549 human lung cancer cell line

目的观察廿烷五烯酸（EPA）联合顺铂对肺腺癌细胞株A-549的抗增殖和诱导凋亡作用。方法A-549进行体外培养后分为A组和B组。B组分别加EPA15（B1组）、30（B2组）、60（B3组）μg/ml及顺铂30μg/ml（B4组）、顺铂30μg/ml＋EPA15μg/ml（B5组）、顺铂30μg/ml＋EPA 30μg/ml（B6组）、顺铂30μg/ml＋EPA60μg/m（B7组）,分别培养24、48、72、96 h。A组加培养液50μl。实验步骤两组相同。MTT法测A-549的生长抑制率,倒置显微镜、光学显微镜观察细胞形态,TUNEL法检测细胞凋亡率。结果①B1～B7（EPA在15～60μg/ml范围内）组,A-549均呈抑制生长,EPA与顺铂具有协同抗癌作用,并表现出浓度、时间依赖性关系。②光镜下B组A-549体积变小、变圆,核染色质浓缩或染色质块形成,部分细胞核脱落,细胞膜起泡形成凋亡小体;A组细胞生长旺盛,呈不规则形,核分裂象多见,细胞核规整,染成均一蓝色。倒置显微镜下,B组A-549体积变小、变圆,核染色质凝集,细胞间连接疏松,贴壁能力减弱;A组细胞贴壁生长良好,充分生长,透亮度好。③B1～B3组细胞凋亡指数（AI）依次升高，分别为5．57％±0．82％、11．68％±1．26％、24．53％±1．68％（P〈0．01）。结论体外实验中，EPA对A-549生长有抑制作用，呈明显的量效和时效关系；EPA与顺铂有协同作用。

Objective To observe the effects of anti-proliferation and inducing apoptosis on A-549 cell line induced by eicosapentaenoic acid（EPA）and its synergistic effect with cisplatin（DDP）.Methods A-549 cell line was cultured in vitro.A-549 cells were divided into group A and group B,the group B was divided into EPA 15（B1 group）,30（B2 group）,60（B3 group）,DDP 30 μg/ml（B4 group）,DDP30 μg/ml ＋EPA 15 μg/ml（B5 group）,DDP 30 μg/ml ＋EPA 30 μg/ml（B6 group） and DDP 30 μg/ml＋EPA 60 μg/ml（B7 group）.The group A added control fluid 500 μl to replace the drugs, the experiment steps were alike. MTT assay was used to determine the effect of EPA on A-549 cells growth inhibitory. Morphological changes of the apoptosis of A-549 cells were observed by using inverted microscope. TUNEL was used to detect cell apoptotie index of A-549. Results ①in the group B1 - B7 ,the growth of A-549 cell were inhibited and in dose and time dependent manner. EPA combined with DDP shows significant synergistic anti-tumor effect on A-549 ceils. ②The A-549 cells in gToup B showed morphological ehanges such as cellular volume decreased and become round, nuclear ehromatin condensed plasma membrance hleb formed, also the apoptotie body were observed by microscope nuclear chromatin condensation ,loose between the cells and adherent ability decrease under invert microscope. ③The apoptotie index （AI） of A-549 ceils in group B1 - B3 was 5.57% ±0.82%, 11.68% ±1.26%, 24.53%± 1.68% ,respectively,there was staffstie significance compared with different concentration （ P 〈 0.01 ）. Conduslons EPA inhibited growth of A-549 cell line in vitro in dose and time dependent manner,EPA combined with DDP shows significant synergistic anti-tumor effect on A- 549 cell line.