新型八核铜配合物乳剂的抗肿瘤作用研究
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摘要
目的：研究新型八核钢配合物乳剂(N-Cu)的体内外抗肿瘤作用及其对S180细胞周期的影响。
方法：1.采用MTT法测定新型八核铜配合物乳剂(N-Cu)对4株人肿瘤细胞,人结肠癌细胞(HT29),人宫颈癌细胞(Hela),人胃癌细胞(SGC7901)和人食管癌细胞(EC9706))作用24 h后对细胞增值的影响。统计得出新型八核铜配合物乳剂对不同肿瘤细胞的抑制率和半数抑制浓度(IC50)。
2.将S180腹水瘤小鼠随机分组,尾静脉注射低、中、高三个浓度的新型八核铜配合物乳剂(N-Cu),观察其不同浓度对小鼠体重、采食量、腹围、腹水体积、生命延长率、血液生化及主要脏器的影响。
3.在前一章动物试验的基础上用流式细胞仪测定新型八核铜配合物乳剂作用S180细胞24 h后的细胞周期分布。结果：1.由MTT试验结果可知：N-Cu分别作用四种肿瘤细胞24 h后,细胞增殖均受到了不同程度的抑制。且随着药物浓度的增大,抑制作用明显增强。且在药物浓度为100.00μg/ml时,抑制率都达到了70%以上。N-Cu作用HT29、Hela、SGC7901和EC9706细胞24 h的半数抑制浓度(IC50)分别为：23.50±0.32,17.50±0.02,34.50±1.04,31.95±0.51μg/ml。
2.动物体内试验结果显示：N-Cu对荷瘤小鼠的体重及采食量无明显抑制作用,能明显减小荷瘤小鼠的腹围及腹水量,且生命延长率有明显的延长。病理切片结果显示高剂量组小鼠的肝脏有明显好转。N-Gu对小鼠的主要脏器无明显的毒副作用。
3.细胞周期试验结果显示：N-Cu能使G0/G1期细胞明显增多,S期细胞减少,细胞被阻滞于G0/G1期。其中高剂量组作用最明显,G0/G1期细胞达到76.1%,中剂量组达到72.3%,差异具有统计学意义(p<0.05)。
结论：新型八核铜配合物乳剂对于四种肿瘤细胞的增殖均有明显的抑制作用。且随着药物浓度的增大,抑制作用明显增强；动物体内抗肿瘤作用较好且无明显的毒副作用；流式细胞仪测定细胞周期结果显示N-Cu具有抗S180细胞增殖的作用,能使G0/G1期细胞明显增多,S期细胞减少,细胞被阻滞于G0/G1期。说明N-Cu是一种具有广阔开发前景的抗肿瘤药物。
Objective:To study the anti-tumor effect of a new eight-copper complex emulsion (N-Cu) using MTT test in vitro and a Sarcoma180 tumor (S180) animal model in vivo, and flow cytometry.
Methods:1.To determine the effects of new eight-copper complex emulsion (N-Cu) on proliferation of four human tumor cell lines, human colon cancer cells (HT29), human cervical carcinoma cells (Hela), human gastric cancer cells (SGC7901)and human esophageal cancer cells (EC9706) using MTT assay after 24 h. Then by statistics obtain the inhibition of tumor cells and the half inhibition concentration (IC50) of new eight-copper complex emulsion.
2. To randomize the mice and inject the new eight-copper complex emulsion (N-Cu) by tail vein of low, medium and high concentrations in mice with S180 ascites-tumor, observing its effects on body weight, feed intake, abdominal circumference, ascites volume, life extension rate, blood biochemistry and major organs.
3. To carry out cell cycle analysis by flow cytometry based on animal experiments in the previous chapter after 24 h.
Results:1.The MTT result showed that N-Cu significantly inhibited the proliferation of the four tumor cell lines after 24 h. With increasing drug concentration, the inhibition was significantly enhanced. And when the drug concentration is 100.00μg/ml, the inhibitory rate reached 70% or more.The IC50s of N-Cu in HT29, Hela, SGC7901 and EC9706 cells after 24 h were 23.50±0.32,17.50±0.02,34.50±1.04, and 31.95±0.51μg/ml, respectively.
2. The results in vivo showed that N-Cu had no significant inhibitory effect on body weight and food intake in mice. However, N-Cu significantly reduced the abdominal circumference and ascites of tumor-bearing mice, and prolonged life span without apparent side effects. Biopsy showed the liver in high dose group had significant improvement in mice.
3. S180 cells treated with N-Cu showed an increase in G0/G1 phase and decrease in S phase. Cells were arrested in G0/G1 phase. Which the most significant effect is high dose group, the percentages of G0/G1 phase cells were increased to 76.1% and 72.3% in the high and middle dose groups, respectively. The difference was statistically significant (p<0.05).
Conclusion:N-Cu significantly inhibited the proliferation of the four tumor cell lines. With increasing drug concentration, the inhibition was significantly enhanced. The results in vivo showed that N-Cu had better anti-tumor effects with no obvious side effects; S180 cells treated with N-Cu by flow cytometry showed that N-Cu with anti-S180 cell proliferation, an increase in G0/G1 phase and decrease in S phase. Cells were arrested in G0/G1 phase. That N-Cu is a broad development prospect of the anti-tumor drugs.
方法：1.采用MTT法测定新型八核铜配合物乳剂(N-Cu)对4株人肿瘤细胞,人结肠癌细胞(HT29),人宫颈癌细胞(Hela),人胃癌细胞(SGC7901)和人食管癌细胞(EC9706))作用24 h后对细胞增值的影响。统计得出新型八核铜配合物乳剂对不同肿瘤细胞的抑制率和半数抑制浓度(IC50)。
2.将S180腹水瘤小鼠随机分组,尾静脉注射低、中、高三个浓度的新型八核铜配合物乳剂(N-Cu),观察其不同浓度对小鼠体重、采食量、腹围、腹水体积、生命延长率、血液生化及主要脏器的影响。
3.在前一章动物试验的基础上用流式细胞仪测定新型八核铜配合物乳剂作用S180细胞24 h后的细胞周期分布。结果：1.由MTT试验结果可知：N-Cu分别作用四种肿瘤细胞24 h后,细胞增殖均受到了不同程度的抑制。且随着药物浓度的增大,抑制作用明显增强。且在药物浓度为100.00μg/ml时,抑制率都达到了70%以上。N-Cu作用HT29、Hela、SGC7901和EC9706细胞24 h的半数抑制浓度(IC50)分别为：23.50±0.32,17.50±0.02,34.50±1.04,31.95±0.51μg/ml。
2.动物体内试验结果显示：N-Cu对荷瘤小鼠的体重及采食量无明显抑制作用,能明显减小荷瘤小鼠的腹围及腹水量,且生命延长率有明显的延长。病理切片结果显示高剂量组小鼠的肝脏有明显好转。N-Gu对小鼠的主要脏器无明显的毒副作用。
3.细胞周期试验结果显示：N-Cu能使G0/G1期细胞明显增多,S期细胞减少,细胞被阻滞于G0/G1期。其中高剂量组作用最明显,G0/G1期细胞达到76.1%,中剂量组达到72.3%,差异具有统计学意义(p<0.05)。
结论：新型八核铜配合物乳剂对于四种肿瘤细胞的增殖均有明显的抑制作用。且随着药物浓度的增大,抑制作用明显增强；动物体内抗肿瘤作用较好且无明显的毒副作用；流式细胞仪测定细胞周期结果显示N-Cu具有抗S180细胞增殖的作用,能使G0/G1期细胞明显增多,S期细胞减少,细胞被阻滞于G0/G1期。说明N-Cu是一种具有广阔开发前景的抗肿瘤药物。
Objective:To study the anti-tumor effect of a new eight-copper complex emulsion (N-Cu) using MTT test in vitro and a Sarcoma180 tumor (S180) animal model in vivo, and flow cytometry.
Methods:1.To determine the effects of new eight-copper complex emulsion (N-Cu) on proliferation of four human tumor cell lines, human colon cancer cells (HT29), human cervical carcinoma cells (Hela), human gastric cancer cells (SGC7901)and human esophageal cancer cells (EC9706) using MTT assay after 24 h. Then by statistics obtain the inhibition of tumor cells and the half inhibition concentration (IC50) of new eight-copper complex emulsion.
2. To randomize the mice and inject the new eight-copper complex emulsion (N-Cu) by tail vein of low, medium and high concentrations in mice with S180 ascites-tumor, observing its effects on body weight, feed intake, abdominal circumference, ascites volume, life extension rate, blood biochemistry and major organs.
3. To carry out cell cycle analysis by flow cytometry based on animal experiments in the previous chapter after 24 h.
Results:1.The MTT result showed that N-Cu significantly inhibited the proliferation of the four tumor cell lines after 24 h. With increasing drug concentration, the inhibition was significantly enhanced. And when the drug concentration is 100.00μg/ml, the inhibitory rate reached 70% or more.The IC50s of N-Cu in HT29, Hela, SGC7901 and EC9706 cells after 24 h were 23.50±0.32,17.50±0.02,34.50±1.04, and 31.95±0.51μg/ml, respectively.
2. The results in vivo showed that N-Cu had no significant inhibitory effect on body weight and food intake in mice. However, N-Cu significantly reduced the abdominal circumference and ascites of tumor-bearing mice, and prolonged life span without apparent side effects. Biopsy showed the liver in high dose group had significant improvement in mice.
3. S180 cells treated with N-Cu showed an increase in G0/G1 phase and decrease in S phase. Cells were arrested in G0/G1 phase. Which the most significant effect is high dose group, the percentages of G0/G1 phase cells were increased to 76.1% and 72.3% in the high and middle dose groups, respectively. The difference was statistically significant (p<0.05).
Conclusion:N-Cu significantly inhibited the proliferation of the four tumor cell lines. With increasing drug concentration, the inhibition was significantly enhanced. The results in vivo showed that N-Cu had better anti-tumor effects with no obvious side effects; S180 cells treated with N-Cu by flow cytometry showed that N-Cu with anti-S180 cell proliferation, an increase in G0/G1 phase and decrease in S phase. Cells were arrested in G0/G1 phase. That N-Cu is a broad development prospect of the anti-tumor drugs.
引文
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