海巴戟果实多糖的体外抗肿瘤活性研究

Antitumor activity in vitro of polysaccharide from Morinda citrifolia L.fruit

【目的】明确海巴戟果实多糖的抗肿瘤活性,为深入研究海巴戟果实多糖生物活性及其开发利用提供科学依据。【方法】采用噻唑蓝比色(MTT)法检测海巴戟果实粗多糖和纯化多糖对人正常结肠上皮细胞的毒性,并鉴定多糖对人结肠癌细胞SW620和HCT-116、宫颈癌细胞Hela、肝癌细胞QGY-7703增殖生长的影响,通过统计癌细胞的凋亡和坏死频率评估纯化多糖的体外抗肿瘤效果。【结果】海巴戟果实多糖无毒,较低浓度(62.5~125μg/mL)的粗多糖或较高浓度(125~2000μg/mL)的纯化多糖对人正常结肠上皮细胞增殖有促进作用。海巴戟果实多糖对宫颈癌细胞Hela的抑制作用较强,粗多糖和纯化多糖均在125~2000μg/mL浓度范围内显著抑制宫颈癌细胞Hela的增殖生长(P<0.05,下同),粗多糖和纯化多糖对宫颈癌细胞Hela的半抑制浓度(IC_(50))分别为542.80和1546.00μg/mL。海巴戟果实多糖对结肠癌细胞HCT-116的增殖生长也有较强抑制作用,但对肝癌细胞QGY-7703的增殖无明显抑制作用。海巴戟果实粗多糖浓度为1000μg/mL时对结肠癌细胞HCT-116和宫颈癌细胞Hela的抑制作用最强,细胞增殖率分别为78.11%和81.85%;纯化多糖浓度为2000μg/mL时对结肠癌细胞HCT-116和宫颈癌细胞Hela的抑制作用最强,细胞增殖率分别为57.05%和68.64%;纯化多糖对癌细胞生长的抑制能力整体上高于粗多糖。纯化多糖对结肠癌细胞HCT-116和宫颈癌细胞Hela的凋亡及坏死频率均有显著影响,且2种癌细胞的坏死频率均随纯化多糖浓度的增加而升高,于2000μg/mL浓度时达峰值,坏死频率分别为2.83%和6.75%。【结论】海巴戟果实多糖具有良好的抗肿瘤活性,纯化多糖比粗多糖的抗肿瘤活性更强。

【Objective】To clarify the antitumor activity of polysaccharides from the Morinda citrifolia L.fruit,and to provide reference basis for the in-depth study of polysaccharide biological activity in M.citrifolia fruit and its development and application.【Method】The cytotoxicity of crude and purified polysaccharides of M.citrifolia fruit on normal colon epithelial cells was detected by thiazole blue colorimetry(MTT)assay.Then,the effects of polysaccharides on the proliferation and growth of human colon cancer cells SW620 and HCT-116,cervical cancer cell Hela and liver cancer cell QGY-7703 were evaluated,and the antitumor activity in vitro of purified polysaccharide was assessed by the frequencies of apoptosis and necrosis of cancer cells.【Result】The polysaccharides of M.citrifolia fruit were non-toxic,and crude polysaccharide at low concentrations(62.5-125μg/mL)or purified polysaccharide at high concentrations(125-2000μg/mL)had promoting effect on the proliferation of human normal colon epithelial cells.Polysaccharide from M.citrifolia fruit showed the strongest inhibitory effect on cervical cancer cell Hela,with both crude and purified polysaccharide significantly inhibiting the growth of cervical cancer cell Hela at the concentration range of 125-2000μg/mL(P<0.05,the same below),with semi-inhibitory concentration(IC_(50))of 542.80 and 1546.00μg/mL.The polysaccharide of M.citrifolia fruit also had strong inhibitory effect on the proliferation and growth of colon cancer cell HCT-116,but had no obvious inhibitory effect on the proliferation of liver cancer cell QGY-7703.At a concentration of 1000μg/mL,the crude polysaccharide of M.citrifolia fruit had the strongest inhibitory effect on colon cancer cells HCT-116 and cervical cancer cells Hela,the cell proliferation rates were 78.11%and 81.85%respectively.The purified polysaccharide at a concentration of 2000μg/mL showed the strongest inhibitory effect on colon cancer cells HCT-116 and cervical cancer cells Hela,with cell proliferation rates of 57.05%and 68.64%.The growth inhibitory ability of purified polysaccharide on cancer cells was higher than that of crude polysaccharide.The purified polysaccharide had significant effect on the apoptosis and necrosis frequency of colon cancer cell HCT-116 and cervical cancer cell Hela,and the necrosis frequency of the both cancer cells increased with the increase of purified polysaccharide concentration,reaching a peak value at 2000μg/mL,with the necrosis frequency of 2.83%and 6.75%.【Conclusion】The polysaccharide from M.citrifolia fruit has good activity of antitumor,and the purified polysaccharide shows much stronger antitumor activity than the crude polysaccharides.