二十碳五烯酸协同长春瑞滨对人肺癌A-549细胞的作用

[目的]探讨二十碳五烯酸(EPA)联合长春瑞滨(NVB)对人肺癌A-549细胞株的增殖、凋亡及细胞周期的影响.[方法]选用人肺癌A-549细胞株进行体外培养.应用倒置显微镜、HE染色、AnnexinⅤ-PI双染色荧光显微镜观察细胞凋亡形态学变化.应用流式细胞仪检测EPA和NVB联合作用于人肺癌A-549细胞株后细胞凋亡情况.[结果]在倒置显微镜下,用EPA处理的A-549肿瘤细胞体积变小、变圆,核染色质凝集,细胞膜起泡形成凋亡小体,细胞间连接疏松,贴壁能力减弱.HE染色:在倒置显微镜下,EPA处理组A-549肿瘤细胞体积变小、变圆,核染色质浓缩或染色质块形成,有的细胞核脱落,有的细胞膜起泡形成凋亡小体.MTT实验结果表明,EPA在15～45 mg/L范围内,于24,48,72h对肺癌A-549细胞株均有抑制生长作用且与NVB有协同作用,并表现出质量浓度、时间依赖性关系.在An-nexinⅤ-PI双染色荧光显微镜下,实验组A-549肿瘤细胞核染色质呈红色或A-549肿瘤细胞呈绿色,细胞周围可见亮绿色荧光的凋亡小体.流式细胞仪检测结果发现,EPA联合NVB组的凋亡率较单用NVB组明显增加.[结论]EPA与NVB对人肺癌A-549细胞的抑制具有协同作用,与单用NVB相比,联合应用可减少NVB的用药剂量.

Effects of eicosapentaenoic acid and its synergistic effects with vinorelbine on A-549 human lung cancer cell line

OBJECTIVE To study the effects of eicosapentaenoic acid(EPA) and its synergistic effects with the vinorelbine(NVB) on the proliferation,apoptosis and cell cycle of A-549 human lung cancer cells line(A-549 HLCCs).METHODS A-549 human lung cancer cells was cultured in vitro,and its morphological changes of apoptosis were observed by using inverted light microscope,HE and AnnexinⅤ-PI double staining with fluorescence microscope.The apoptosis of A-549 HLCCs exposed to EPA and its synergistic effects with NVB was analyzed by flow cytometry.RESULTS The A-549 HLCCs treated with EPA by HE staining showed the small and round cellular volume,nuclear chromatin condensation,bleb formation of cell membrane,formations of apoptotic body,loose of intercellular connection and decrease of adherent ability decrease under invert and light microscopes,respectively.The MTT test showed that when 15-45mg/L of EPA exposed for 24 h,48 h and 72 h,the growth of A-549 HLCCs was inhibited,and EPT had synergistic effects with NVB,and showed in dose and time dependent manner.EPA combined with NVB showed significant synergistic anti-tumor effect on A-549 HLCCs.As stained with AnnexinⅤ-PI double fluorescent staining and observed under fluorescent microscope,the A-549 HLCCs appeared as chromatine red condensation in nuclear or green condensation in cells,and light green fluorescent apoptotic bodies around cells.Apoptotic rate in group of EPA combined with NVB increased significantly than that in NVB group by flow cytometry.CONCLUSION EPA and NVB have synergetic effects in the inhibition of A-549 HLCCs,and the combination of NVB and EPA can reduce the dosage of NVB as compared with NVB monotherapy.