海带多糖对人鼻咽癌细胞及其裸鼠移植瘤的抑制作用
Inhibition of Human Nasopharyngeal Carcinoma Cell Proliferation and Xenograft Growth in Nude Mice by Laminaria Japonica Polysaccharides
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摘要:目的 通过观察海带多糖(laminaria japonica polysaccharides, LJP)在体内外对人鼻咽癌(nasopharyngeal carcinoma, NPC)细胞的抑制作用, 探讨其可能的抗癌机制。方法 应用MTT法检测LJP对人NPC细胞株(HONE1和CNE2)增殖的抑制作用; 运用PI加Annexin V双染法检测LJP对HONE1细胞凋亡的影响; 以人NPC细胞株HONE1建立裸鼠皮下移植瘤模型及行体内抑瘤实验, 并在透射电镜(transmission electron microscopy, TEM)下观察移植瘤细胞超微结构。结果 MTT显示LJP对人NPC细胞(HONE1和CNE2)的增殖有抑制作用, 且具有剂量相关性, 320 mg/L的LJP作用72h的抑制率分别为58.27%(P < 0.01)和55.00%(P < 0.01);流式细胞仪检测结果显示, LJP具有诱导HONE1细胞凋亡的作用, 凋亡率随着LJP浓度的增加而增高, 320mg/L的凋亡率为(49.51±1.89)%(P < 0.01)。体内实验中, LJP中、高剂量组抑瘤率分别为33.7%(P < 0.05)和47.0%(P < 0.01), 具有明显抑制移植瘤的作用, 而低剂量组的抑瘤率仅为16.4%(P > 0.05)。TEM结果提示癌细胞呈现凋亡特征的改变。结论 LJP具有抑制NPC细胞生长的作用, 机制可能是通过诱导癌细胞凋亡而实现的。Abstract:Objective This study aims to evaluate the antitumor activity of Laminaria japonica polysaccharides(LJP) on human nasopharyngeal carcinoma(NPC) cells in vitro and in vivo, and to explore the underlying mechanisms.Methods Different human NPC cell lines were treated with LJP.and cell proliferation was examined by MTT assays.The cell apoptosis of LJP-treated HONE1 was examined by double staining assay.A tumor model established by the subcutaneous inoculation of NPC cell HONE1 into nude mice was used to evaluate the inhibitory action of LJP in vivo.Transmission electron microscopy(TEM) was used to observe changes in the ultrastructure of the NPC cells in xenografts.Results The anti-proliferative activity of LJP was found in both human NPC cell lines(HONE1 and CNE2) by MTT assay.Flow cytometry showed that LJP can induce the apoptosis of HONE1.With increased concentration of LJP.the apoptosis rate increased.The apoptosis rate was 49.51%±1.89%(P < 0.01) when treated with 320 mg/L LJP. The inhibition ratio was between 50%and 60%at 72 h after treatment with 320 mg/L LJP.Compared with the control group, the growth of xenografts in nude mice was significantly suppressed following the administration of LJP in a dose-dependent manner.The inhibition ratios were 33.7%(P < 0.05) and 47.0%(P < 0.01) when treated with 25 mg/kg and 50 mg/kg LJP.respectively.However, the inhibition ratio of the 12.5 mg/kg group was only 16.4%(P > 0.05).The typical morphological changes in apoptosis were revealed by TEM.Conclusion LJP can inhibit the growth of NPC probably by inducing the apoptosis of NPC cells in vitro and in vivo.
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