【作者】 张丹华；

【导师】 周恩相；

【作者基本信息】 中南大学 ， 外科学， 2010， 硕士

【摘要】 目的：研究蜈蚣提取液对乳腺癌细胞株的敏感性,为乳腺癌的临床治疗提供理论依据。方法：体外培养乳腺癌细胞株MDA-MB-231,采用不同浓度蜈蚣提取液(Extract of Centipede, ECP)予以干预,通过MTT实验、细胞的药物浓度-时间生长曲线研究其对乳腺癌细胞株的敏感性；利用倒置光学显微镜和电子显微镜观察细胞形态及其超微结构的变化。结果：①MTT法揭示各浓度ECP对人乳腺癌细胞株MDA-MB-231生长均有抑制作用。ECP在80mg/ml、40 mg/ml、20 mg/ml、10mg/ml、5 mg/ml、2.5 mg/ml及1.25 mg/ml下对MDA-MB-231细胞的抑制率依次为(71.26±2.64)%、(62.48±6.67)%、(56.26±3.17)%、(48.63±4.48)%、(42.13±4.73)%、(34.5±2.98)%和(26.89±2.95)%,各组间有显著差异性(P<0.01); ECP在80mg/ml浓度组的抑制率与吡柔比星(THP) 10μg/ml抑制率比较无统计学差异(P>0.05),大致相同；ECP在20mg/ml及40mg/ml组的抑制率与THP5μg/ml抑制率比较无统计学差异(P>0.05),大致相同；由公式计算得出的中效浓度为12.5mg/ml。②细胞生长曲线图显示MDA-MB-231活细胞数目与ECP浓度及药物作用时间相关。治疗组浓度在12.5mg/ml与25mg/ml时,用药后24小时活细胞数就显著减少。各组间活细胞数比较差异明显(P<0.01)。③光镜下见ECP作用48小时后,浓度低于10mg/ml对MDA-MB-231细胞抑制作用不明显,浓度在10mg/ml以上时,MDA-MB-231活细胞数减少,细胞形态变圆变小,80mg/ml时见细胞大片坏死,有较多的不规则细胞碎片,培养瓶中坏死脱落细胞增多。电镜下见治疗组细胞密度增高,染色质凝集固缩,染色质边集,核固缩、核碎裂,及凋亡小体的形成。结论：①ECP能抑制人乳腺癌细胞株MDA-MB-231的增殖,且抑制作用呈剂量和时间依赖效应；②ECP低浓度有诱导MDA-MB-231细胞凋亡作用,中、高浓度有直接杀伤MDA-MB-231细胞作用。更多还原

【Abstract】 Objective:To investigate sensitivity of the effect of centipede extract on breast cancer cell lines and provide evidences for further clinic therapy of breast cancerMethods:Cultured breast cancer cells MDA-MB-231 are intervened with different concentration of centipede extract. MTT assay and concentration-time curve are used to study the effect sensitivity of centipede extract on breast cancer cell lines; Inverted light microscope and electron microscope are used to observe cell changes in morphology and ultrastructure.Results:①The results of MTT assay showed that different concentrations of centipede extract all can inhibit the growth of human breast cancer cells MDA-MB-231.The inhibition rates were (71.26±2.64)%, (62.48±6.67)%, (56.26±3.17)%, (48.63±4.48)%, (42.13±4.73)%, (34.5±2.98)% and (26.89±2.95)% respectively,while the Corresponding concentration of centipede extract were 80mg/ml,40 mg/ ml,20 mg/ml, 10mg/ml,5 mg/ml,2.5 mg/ml and 1.25 mg/ml,and each group had significant differences(P<0.01). ECP concentrations in 80mg/ml inhibition rate of pirarubicin (THP) 10μg/ml was no significant difference in inhibition rate(P>0.05), roughly the same, ECP in the inhibition of 20mg/ml and 40mg/ml group rate and THP5μg/ml no significant difference in inhibition rate (P>0.05), roughly the same. IC50 of centipede extract to MDA-MB-231 was 12.5mg/ml.②Drug concentration-time curve revealed that the viable cell counts of MDA-MB-231 has direct relations with the concentration and administrative time of centipede extract. When treated with 12.5mg/ml and 25mg/ml centipede extract, viable counts of MDA-MB-231 sharply decrease after 24 hours treatment and the difference among treatment groups is significant (P<0.01).③After 48hr intervention, there were no inhibitory effect observed by light microscopy in groups with concentration less than 10mg/ml; viable counts of MDA-MB-231 decreased and the cells transformed to round and small when concentration were more than 10mg/ml; With treated with 80mg/ml, large number of necrotic cells and irregular cell debris were seemed. Meanwhile, margination and condensation of chromatin, fragmentation and condensation of nucleus and formation of apoptotic bodies were found in cells by electron microscope which had higher density.Conclusion:①Extract of centipede can inhibit the growing of human breast cancer cells MDA-MB-231 and the effect was concentration-time dependent;②Extract of centipede can induce MDA-MB-231 cell line to apoptosis in low concentration and directly kill it in middle or high concentration.更多还原