【作者】 殷玉琨；

【导师】 宋爱莉；

【作者基本信息】 山东中医药大学 ， 中医外科学， 2013， 博士

【摘要】 目的：通过研究榄香烯乳注射液作用于乳腺癌及血管内皮细胞株功能改变及作用机制的体外研究，观察其对乳腺癌及血管内皮细胞株的生物学行为影响，并检测对“三阴性乳腺癌”细胞MDA-MB-231的凋亡诱导。方法：设立空白对照组，将不同浓度榄香烯乳作用于乳腺癌细胞系MDA-MB-231及MCF-7细胞作为处理组，结晶紫实验检测细胞生长速率，ECIS实验检测细胞粘附功能及迁徙功能，侵袭实验检测细胞侵袭能力。作用于血管内皮细胞系HECV，结晶紫实验检测细胞生长速率，ECIS实验检测细胞粘附功能及迁徙功能，新生血管形成实验检测血管生成能力，细胞聚集实验检测细胞聚集能力。榄香烯乳作用于MDA-MB-231细胞作为处理组，同时设立空白对照组，倒置显微镜观察细胞形态，实时定量PCR检测细胞凋亡指标Caspase3、Caspase8、Caspase9的表达。结果：榄香烯达到一定浓度时，能够有效的抑制MDA-MB-231（浓度≥20μg/mL时，P<0.05）及MCF-7（浓度=160μg/mL时，P<0.05）细胞的生长速率，且呈剂量依赖性；随着榄香烯乳浓度的增加，MDA-MB-231（浓度≥80μg/mL时，P<0.05）及MCF-7（浓度=160μg/mL时，P<0.05）的细胞的粘附能力及MDA-MB-231（浓度≥40μg/mL时，P<0.05）的细胞迁徙能力减低，但对MCF-7细胞的迁徙能力改变不明显；榄香烯浓度为160μg/mL时，MCF-7的侵袭率降低（P<0.05），而MDA-MB-231的侵袭率无统计学差异。低浓度的榄香烯乳可以促进HECV细胞的生长速率（=20μg/mL时，P<0.05）、粘附能力（=10μg/mL时，P<0.05）、迁徙能力（=10μg/mL时，P<0.05）及新生血管形成能力（=20μg/mL时，P<0.05）；而高浓度的榄香烯乳则抑制其生长速率（≥80μg/mL时，P<0.05）、粘附能力（≥80μg/mL时，P<0.05）、迁徙能力（≥80μg/mL时，P<0.05）、新生血管形成能力（≥80μg/mL时，P<0.05）及细胞聚集能力（=160μg/mL，90min时，P<0.05）。榄香烯乳浓度为40μg/mL时，对比于对照组，镜下观察细胞的形态呈现典型的凋亡改变；MDA-MB-231细胞凋亡因子Caspase3（56.7±15.7vs.812.2±222.6，P=0.025）、Caspase8（140.3±25.6vs.327.8±50.1，P=0.032）表达升高，Caspase9（13.1±4.4vs.5.7±1.5，P=0.623）表达无统计学差异。结论：榄香烯乳注射液具有抑制乳腺癌细胞生长、粘附、迁移及侵袭的作用。而对于血管内皮细胞，低浓度的榄香烯乳注射液对其细胞功能具有促进作用，而高浓度的榄香烯乳则表现为抑制作用。榄香烯乳可以诱导乳腺癌细胞株MDA-MB-231的细胞凋亡。更多还原

【Abstract】 Objective: The aim of this study is to test whether Elemene has the ability to impactthe functions of breast cancer cell lines of MDA-MB-231and MCF-7and endothelial cellline HECV, and to find the mechanism.Methods: After treated with Elemene as the treatment group and compared withcontrol group, the growth rates, adhesion abilities, migration abilities and invasion abilitiesof MDA-MB-231and MCF-7cells were successively detected by Crystal violetexperiment, ECIS assay and Invasion assay.After treated with Elemene as the treatment group and compared with control group,the growth rates, adhesion abilities, migration abilities, tubule formation and cellaggregation of HECV cells were successively detected by Crystal violet experiment, ECISassay, tubule formation assay and cell aggregation assay.MDA-MB-231cells were treated by Elemene at a concentration of40μg/mL for48hours. Morphological changes were observed by Inverted microscope, and Caspase-3,-8,-9RNA levels were investigated by Q-PCR.Results: When Elemene reached on a certain concentration, the growth rates ofMDA-MB-231(≥20μg/mL, P<0.05) and MCF-7(=160μg/mL, P<0.05) cells wereinhibited, and had a dose-dependent. As the increasing of the concentration of Elemene,the adhesion ability of MDA-MB-231cells (≥80μg/mL, P<0.05) and MCF-7cells(=160μg/mL, P<0.05) were down-regulated; The migration abilities of MDA-MB-231cells were reduced (≥40μg/mL, P<0.05), while the changes of MCF-7cells showed no statisticaldifference; The invasion ability of MCF-7cells were impacted by Elemene (=160μg/mL,P<0.05), but the same phenomenon did not appear on MDA-MB-231cells.Elemene at a low concentration has the ability to enhance HECV cells on growth(=20μg/mL, P<0.05), adhesion (=10μg/mL, P<0.05), migration (=10μg/mL, P<0.05) andtubule formation (=20μg/mL, P<0.05), while the high concentration of Elemene inhibitsthe functions of growth (≥80μg/mL, P<0.05), adhesion (≥80μg/mL, P<0.05), migration(≥80μg/mL, P<0.05), tubule formation (≥80μg/mL, P<0.05) and cell aggregation(≥80μg/mL, P<0.05).Cells were changed to an apoptotic status under the microscope, and the expressionlevels of Caspase (Caspase3:56.7±15.7vs.812.2±222.6, P=0.025; Caspase8:140.3±25.6vs.327.8±50.1, P=0.032; Caspase9:13.1±4.4vs.5.7±1.5, P=0.623) of MDA-MB-231cellswere upgraded after treated with Elemene at a concentration of40μg/mL.Conclusion: Elemene has the capability to inhibit the growth rates, adhesion ability,migration ability and invasion ability of breast cancer cells. While for endothelial cells,Elemene has the capability to enhance cell functions at a low concentration, but inhibits thefunctions at high concentration. Elemene has the capability to induct apoptosis of breastcancer cell line MDA-MB-231.更多还原