榄香烯乳注射液对乳腺癌及血管内皮细胞功能改变及作用机制的体外研究
|
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。
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.
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。
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.
引文
[1]郑荣寿,张思维,吴良有,等.中国肿瘤登记地区2008年恶性肿瘤发病和死亡分析[J].中国肿瘤,2012,(01):1-12.
[2] Swain, SM. Triple-negative breast cancer: meta-static risk and role of platinumagents [C].44th Annual Meeting of the American Society of Clinical Oncology.2008.30-June3.
[3]廖仕翀,喻莉,李金芯,等.三阴性乳腺癌的临床特点及治疗进展[J].中华乳腺病杂志（电子版）,2012,06(1):56-59.
[4]许振国,宋爱莉,殷玉琨,等.莪术油对大鼠乳腺癌癌前病变组织VEGF mRNA表达的影响[J].中华肿瘤防治杂志,2012,(08):570-574.
[5]殷玉琨,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变组织中C-erbB-2、P53表达的影响[J].中华中医药学刊,2012,(06):1219-1221+1447.
[6]刘晓菲,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变MVD及相关调控因子VEGF/FLK表达的干预作用研究[J].中医药学报,2011,(04):19-21.
[7] Jianjun Q, Song Z, Yin L, et al. Treatment of chylothorax with elemene[J]. ThoracCardiovasc Surg,2008,56(2):103-5.
[8] Wang B, Peng XX, Sun R, et al. Systematic review of beta-elemene injection asadjunctive treatment for lung cancer[J]. Chin J Integr Med,2012,18(11):813-23.
[9]肖震宇,刘华峰,邓江华.榄香烯注射液联合放疗治疗肺癌脑转移的疗效及对血清基质金属蛋白酶-2和-9的影响[J].中国老年学杂志,2012,32(15):3209-3210.
[10]张丛梅.榄香烯单药或联合放化疗治疗脑恶性肿瘤的分析[J].亚太传统医药,2012,08(3):66-67.
[11]孙鲁江,王乃东.肝动脉灌注并瘤体内注射榄香烯-碘油乳剂治疗肝癌40例[J].中国实用医药,2012,7(22):166-167.
[12]王晋舜.腹腔内注射榄香烯乳与双通路化疗联合治疗癌性腹水临床观察[J].中国实用乡村医生杂志,2012,19(4):51-52.
[13] Lu X, Wang Y, Luo H, et al. beta-elemene inhibits the proliferation of T24bladdercarcinoma cells through upregulation of the expression of Smad4[J]. Mol MedReport,2013,7(2):513-8.
[14] Li QQ, Lee RX, Liang H, et al. Anticancer Activity of beta-Elemene and itsSynthetic Analogs in Human Malignant Brain Tumor Cells[J]. Anticancer Res,2013,33(1):65-76.
[15] Li QQ, Lee RX, Liang H, et al. Enhancement of cisplatin-induced apoptosis bybeta-elemene in resistant human ovarian cancer cells[J]. Med Oncol,2013,30(1):424.
[16] Zhang B, Zhang X, Tang B, et al. Investigation of elemene-induced reversal oftamoxifen resistance in MCF-7cells through oestrogen receptor alpha (ERalpha)re-expression[J]. Breast Cancer Res Treat,2012,136(2):399-406.
[17] Zhang H, Xu F, Xie T, et al. beta-elemene induces glioma cell apoptosis bydownregulating survivin and its interaction with hepatitis B X-interacting protein[J].Oncol Rep,2012,28(6):2083-90.
[18] Jiang ZY, Qin SK, Yin XJ, et al. Synergistic effects of Endostar combined withbeta-elemene on malignant ascites in a mouse model[J]. Exp Ther Med,2012,4(2):277-284.
[19] Zhan YH, Liu J, Qu XJ, et al. beta-Elemene induces apoptosis in human renal-cellcarcinoma786-0cells through inhibition of MAPK/ERK and PI3K/Akt/mTORsignalling pathways[J]. Asian Pac J Cancer Prev,2012,13(6):2739-44.
[20]许振国,宋爱莉,殷玉琨,等.莪术油对大鼠乳腺癌癌前病变组织VEGF mRNA表达的影响[J].中华肿瘤防治杂志,2012,19(8):570-574.
[21]殷玉琨,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变组织中C-erbB-2、P53表达的影响[J].中华中医药学刊,2012,30(06):1219-1221.
[22]李莹,卢海,戴洪雷,等.榄香烯抑制角膜新生血管的实验研究[J].眼科新进展,2004,(04):250-253.
[23]李静蔚,宋爱莉,张敬涛,等. DMBA诱导大鼠乳腺癌癌前病变的组织病理学研究[J].中华中医药学刊,2008,(03):505-508.
[24]李超,伏圣博,刘华玲,等.细胞凋亡研究进展[J].世界科技研究与发展,2007,(03):45-53.
[25]李小艳,包素珍. Caspase家族的研究进展[J].内蒙古中医药,2012,(02):107.
[26]戴昕,李占全,冀林华.凋亡相关蛋白Caspase研究进展[J].中国现代医药杂志,2010,(04):130-132.
[27]宋爱莉,张敬涛,李静蔚,等.莪术油对乳腺癌癌前病变造模大鼠血液流变学及乳房微循环的影响[J].中华中医药学刊,2008,26(3):458-460.
[28]刘晓菲,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变MVD及相关调控因子VEGF/FLK表达的干预作用研究[J].中医药学报,2011,39(4):19-21.
[29]孙子渊,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变组织中Ki67表达的影响[J].中华中医药学刊,2010,(1):48-51.
[30]宋爱莉,殷玉琨,李静蔚,等.莪术油干预治疗DMBA诱导大鼠乳腺癌癌前病变的组织病理学研究[J].中华中医药学刊,2009,27(12):2496-2499.
[1]第二届全国活血化瘀研究学术会议修定·血瘀证诊断标准[J].中西医结合杂志,1987,7(3):129.
[2]孙波.刘尚义教授辨治乳腺癌经验[J].河南中医,2007,(07):13-14.
[3]潘启超，胥彬，等.肿瘤药理学与化学治疗学[M].郑州:河南医科大学,2000:312-315.
[4]张国铎.活血化瘀法抗肿瘤机理的研究进展[J].浙江中医杂志,2002,(04):42-44.
[5]林毅，唐汉钧.现代中医乳房病学[J].人民卫生出版社,2003:318.
[6]王志鹏,张阳.焦中华活血化瘀解毒散结法治疗乳岩1例[J].中国中医药现代远程教育,2010,(02):66.
[7]李静蔚,宋爱莉,张敬涛,等. DMBA诱导大鼠乳腺癌癌前病变的组织病理学研究[J].中华中医药学刊,2008,(03):505-508.
[8]郭少贤,刘永惠,常靖,等.麝香酮干预血瘀证乳腺癌组织血管生成相关因子表达研究[J].河北中医药学报,2011,(02):40-41+2.
[9]宋爱莉，闫珂.复方丹参注射液防治乳腺癌术后并发症的实验研究[J].山东中医药大学学报,2002,26(4):308-311.
[10]郑武，邹荣生.血府逐瘀汤加减结合功能锻炼治疗乳腺癌术后上肢水肿30例[J].福建中医药,2004,35(3):30.
[11]樊晓滨.活血化瘀法治疗乳腺癌根治术后并发症41例[J].国医论坛,2003,(04):28.
[12]刘红梅,卞卫和,任晓梅,等.乳腺癌术后并发症的中医治疗[J].四川中医,2007,(06):85-86.
[13]许正国,孟昭旭.血府逐瘀汤治疗乳腺癌根治术后并发症[J].辽宁中医杂志,2005,(09):947.
[14]崔永安,左小东,毛承飞.活血化瘀法在肿瘤“辨法论治”中的应用[J].中医杂志,2007,(08):749-750.
[15]谢丹,杨海燕,黄梅,等.乳腺癌血瘀状态与新辅助化疗疗效的相关性研究[J].新中医,2007,(06):84-85.
[16]史东星.养阴活血法治疗放射性肺炎急性期临床研究[J].中医学报,2012,(02):139-140.
[17]左益华,钱龙江,王展海,等.活血理气汤配合新辅助化疗治疗乳腺癌32例[J].河南中医,2012,(08):1055-1056.
[18]黄梅.益气活血汤改善脾虚挟瘀型乳腺癌患者化疗毒副作用的临床研究[J].湖北中医杂志,2005,(07):9-10.
[19] Xu HB, Li L, Fu J, et al. Reversion of multidrug resistance in a chemoresistanthuman breast cancer cell line by beta-elemene[J]. Pharmacology,2012,89(5-6):303-12.
[20] Zhang B, Zhang X, Tang B, et al. Investigation of elemene-induced reversal oftamoxifen resistance in MCF-7cells through oestrogen receptor alpha (ERalpha)re-expression[J]. Breast Cancer Res Treat,2012,136(2):399-406.
[21]王辉.益气活血法抗乳腺癌血道转移的基础与临床转化研究[J].中国中医科学院博士论文,2012.
[22]肖丽明，陈健民，许德盛，等.芎龙汤颗粒剂对癌症患者免疫功能影响的观察[J].中成药,1997,19(7):23.
[23]吴新正,何迎春,田道法,等.中药防治癌前病变现状及其机制[J].中草药,2006,(07):1101-1104.
[24]宋爱莉,殷玉琨,李静蔚,等.莪术油干预治疗DMBA诱导大鼠乳腺癌癌前病变的组织病理学研究[J].中华中医药学刊,2009,(12):2496-2499.
[25]殷玉琨,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变组织中C-erbB-2、P53表达的影响[J].中华中医药学刊,2012,(06):1219-1221+1447.
[26]孙子渊,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变组织中Ki67表达的影响[J].中华中医药学刊,2010,(01):48-51.
[27]刘晓菲,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变MVD及相关调控因子VEGF/FLK表达的干预作用研究[J].中医药学报,2011,(04):19-21.
[28]许振国,宋爱莉,殷玉琨,等.莪术油对大鼠乳腺癌癌前病变组织VEGFmRNA表达的影响[J].中华肿瘤防治杂志,2012,(08):570-574.
[29]宋爱莉,张敬涛,李静蔚,等.莪术油对乳腺癌癌前病变造模大鼠血液流变学及乳房微循环的影响[J].中华中医药学刊,2008,(03):458-460.
[30]李振波，丘和明.活血化瘀法治疗癌症的讨论[J].中国中西医结合杂志,1996,16(9):559-561.
[31]韩俊庆，陈延条，满运艳，等.复春片合并放射治疗鼻咽癌临床研究[J].中国中西医结合杂志,1995,15(12):710-712.
[32]张凤瑞.膈下逐瘀汤现代研究概述[J].中成药,1995,17(5):44-45.
[33]李学汤.几种活血化瘀药物对小鼠肝癌细胞形成肺转移影响的初步试验观察[J].中医杂志,1980,21(8):78.
[34]黄孔威.赤芍对实验肿瘤生长和转移的影响及药理作用的研究[J].中华肿瘤杂志,1983,5(1):24.
[2] Swain, SM. Triple-negative breast cancer: meta-static risk and role of platinumagents [C].44th Annual Meeting of the American Society of Clinical Oncology.2008.30-June3.
[3]廖仕翀,喻莉,李金芯,等.三阴性乳腺癌的临床特点及治疗进展[J].中华乳腺病杂志（电子版）,2012,06(1):56-59.
[4]许振国,宋爱莉,殷玉琨,等.莪术油对大鼠乳腺癌癌前病变组织VEGF mRNA表达的影响[J].中华肿瘤防治杂志,2012,(08):570-574.
[5]殷玉琨,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变组织中C-erbB-2、P53表达的影响[J].中华中医药学刊,2012,(06):1219-1221+1447.
[6]刘晓菲,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变MVD及相关调控因子VEGF/FLK表达的干预作用研究[J].中医药学报,2011,(04):19-21.
[7] Jianjun Q, Song Z, Yin L, et al. Treatment of chylothorax with elemene[J]. ThoracCardiovasc Surg,2008,56(2):103-5.
[8] Wang B, Peng XX, Sun R, et al. Systematic review of beta-elemene injection asadjunctive treatment for lung cancer[J]. Chin J Integr Med,2012,18(11):813-23.
[9]肖震宇,刘华峰,邓江华.榄香烯注射液联合放疗治疗肺癌脑转移的疗效及对血清基质金属蛋白酶-2和-9的影响[J].中国老年学杂志,2012,32(15):3209-3210.
[10]张丛梅.榄香烯单药或联合放化疗治疗脑恶性肿瘤的分析[J].亚太传统医药,2012,08(3):66-67.
[11]孙鲁江,王乃东.肝动脉灌注并瘤体内注射榄香烯-碘油乳剂治疗肝癌40例[J].中国实用医药,2012,7(22):166-167.
[12]王晋舜.腹腔内注射榄香烯乳与双通路化疗联合治疗癌性腹水临床观察[J].中国实用乡村医生杂志,2012,19(4):51-52.
[13] Lu X, Wang Y, Luo H, et al. beta-elemene inhibits the proliferation of T24bladdercarcinoma cells through upregulation of the expression of Smad4[J]. Mol MedReport,2013,7(2):513-8.
[14] Li QQ, Lee RX, Liang H, et al. Anticancer Activity of beta-Elemene and itsSynthetic Analogs in Human Malignant Brain Tumor Cells[J]. Anticancer Res,2013,33(1):65-76.
[15] Li QQ, Lee RX, Liang H, et al. Enhancement of cisplatin-induced apoptosis bybeta-elemene in resistant human ovarian cancer cells[J]. Med Oncol,2013,30(1):424.
[16] Zhang B, Zhang X, Tang B, et al. Investigation of elemene-induced reversal oftamoxifen resistance in MCF-7cells through oestrogen receptor alpha (ERalpha)re-expression[J]. Breast Cancer Res Treat,2012,136(2):399-406.
[17] Zhang H, Xu F, Xie T, et al. beta-elemene induces glioma cell apoptosis bydownregulating survivin and its interaction with hepatitis B X-interacting protein[J].Oncol Rep,2012,28(6):2083-90.
[18] Jiang ZY, Qin SK, Yin XJ, et al. Synergistic effects of Endostar combined withbeta-elemene on malignant ascites in a mouse model[J]. Exp Ther Med,2012,4(2):277-284.
[19] Zhan YH, Liu J, Qu XJ, et al. beta-Elemene induces apoptosis in human renal-cellcarcinoma786-0cells through inhibition of MAPK/ERK and PI3K/Akt/mTORsignalling pathways[J]. Asian Pac J Cancer Prev,2012,13(6):2739-44.
[20]许振国,宋爱莉,殷玉琨,等.莪术油对大鼠乳腺癌癌前病变组织VEGF mRNA表达的影响[J].中华肿瘤防治杂志,2012,19(8):570-574.
[21]殷玉琨,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变组织中C-erbB-2、P53表达的影响[J].中华中医药学刊,2012,30(06):1219-1221.
[22]李莹,卢海,戴洪雷,等.榄香烯抑制角膜新生血管的实验研究[J].眼科新进展,2004,(04):250-253.
[23]李静蔚,宋爱莉,张敬涛,等. DMBA诱导大鼠乳腺癌癌前病变的组织病理学研究[J].中华中医药学刊,2008,(03):505-508.
[24]李超,伏圣博,刘华玲,等.细胞凋亡研究进展[J].世界科技研究与发展,2007,(03):45-53.
[25]李小艳,包素珍. Caspase家族的研究进展[J].内蒙古中医药,2012,(02):107.
[26]戴昕,李占全,冀林华.凋亡相关蛋白Caspase研究进展[J].中国现代医药杂志,2010,(04):130-132.
[27]宋爱莉,张敬涛,李静蔚,等.莪术油对乳腺癌癌前病变造模大鼠血液流变学及乳房微循环的影响[J].中华中医药学刊,2008,26(3):458-460.
[28]刘晓菲,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变MVD及相关调控因子VEGF/FLK表达的干预作用研究[J].中医药学报,2011,39(4):19-21.
[29]孙子渊,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变组织中Ki67表达的影响[J].中华中医药学刊,2010,(1):48-51.
[30]宋爱莉,殷玉琨,李静蔚,等.莪术油干预治疗DMBA诱导大鼠乳腺癌癌前病变的组织病理学研究[J].中华中医药学刊,2009,27(12):2496-2499.
[1]第二届全国活血化瘀研究学术会议修定·血瘀证诊断标准[J].中西医结合杂志,1987,7(3):129.
[2]孙波.刘尚义教授辨治乳腺癌经验[J].河南中医,2007,(07):13-14.
[3]潘启超，胥彬，等.肿瘤药理学与化学治疗学[M].郑州:河南医科大学,2000:312-315.
[4]张国铎.活血化瘀法抗肿瘤机理的研究进展[J].浙江中医杂志,2002,(04):42-44.
[5]林毅，唐汉钧.现代中医乳房病学[J].人民卫生出版社,2003:318.
[6]王志鹏,张阳.焦中华活血化瘀解毒散结法治疗乳岩1例[J].中国中医药现代远程教育,2010,(02):66.
[7]李静蔚,宋爱莉,张敬涛,等. DMBA诱导大鼠乳腺癌癌前病变的组织病理学研究[J].中华中医药学刊,2008,(03):505-508.
[8]郭少贤,刘永惠,常靖,等.麝香酮干预血瘀证乳腺癌组织血管生成相关因子表达研究[J].河北中医药学报,2011,(02):40-41+2.
[9]宋爱莉，闫珂.复方丹参注射液防治乳腺癌术后并发症的实验研究[J].山东中医药大学学报,2002,26(4):308-311.
[10]郑武，邹荣生.血府逐瘀汤加减结合功能锻炼治疗乳腺癌术后上肢水肿30例[J].福建中医药,2004,35(3):30.
[11]樊晓滨.活血化瘀法治疗乳腺癌根治术后并发症41例[J].国医论坛,2003,(04):28.
[12]刘红梅,卞卫和,任晓梅,等.乳腺癌术后并发症的中医治疗[J].四川中医,2007,(06):85-86.
[13]许正国,孟昭旭.血府逐瘀汤治疗乳腺癌根治术后并发症[J].辽宁中医杂志,2005,(09):947.
[14]崔永安,左小东,毛承飞.活血化瘀法在肿瘤“辨法论治”中的应用[J].中医杂志,2007,(08):749-750.
[15]谢丹,杨海燕,黄梅,等.乳腺癌血瘀状态与新辅助化疗疗效的相关性研究[J].新中医,2007,(06):84-85.
[16]史东星.养阴活血法治疗放射性肺炎急性期临床研究[J].中医学报,2012,(02):139-140.
[17]左益华,钱龙江,王展海,等.活血理气汤配合新辅助化疗治疗乳腺癌32例[J].河南中医,2012,(08):1055-1056.
[18]黄梅.益气活血汤改善脾虚挟瘀型乳腺癌患者化疗毒副作用的临床研究[J].湖北中医杂志,2005,(07):9-10.
[19] Xu HB, Li L, Fu J, et al. Reversion of multidrug resistance in a chemoresistanthuman breast cancer cell line by beta-elemene[J]. Pharmacology,2012,89(5-6):303-12.
[20] Zhang B, Zhang X, Tang B, et al. Investigation of elemene-induced reversal oftamoxifen resistance in MCF-7cells through oestrogen receptor alpha (ERalpha)re-expression[J]. Breast Cancer Res Treat,2012,136(2):399-406.
[21]王辉.益气活血法抗乳腺癌血道转移的基础与临床转化研究[J].中国中医科学院博士论文,2012.
[22]肖丽明，陈健民，许德盛，等.芎龙汤颗粒剂对癌症患者免疫功能影响的观察[J].中成药,1997,19(7):23.
[23]吴新正,何迎春,田道法,等.中药防治癌前病变现状及其机制[J].中草药,2006,(07):1101-1104.
[24]宋爱莉,殷玉琨,李静蔚,等.莪术油干预治疗DMBA诱导大鼠乳腺癌癌前病变的组织病理学研究[J].中华中医药学刊,2009,(12):2496-2499.
[25]殷玉琨,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变组织中C-erbB-2、P53表达的影响[J].中华中医药学刊,2012,(06):1219-1221+1447.
[26]孙子渊,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变组织中Ki67表达的影响[J].中华中医药学刊,2010,(01):48-51.
[27]刘晓菲,宋爱莉,李静蔚,等.莪术油对大鼠乳腺癌癌前病变MVD及相关调控因子VEGF/FLK表达的干预作用研究[J].中医药学报,2011,(04):19-21.
[28]许振国,宋爱莉,殷玉琨,等.莪术油对大鼠乳腺癌癌前病变组织VEGFmRNA表达的影响[J].中华肿瘤防治杂志,2012,(08):570-574.
[29]宋爱莉,张敬涛,李静蔚,等.莪术油对乳腺癌癌前病变造模大鼠血液流变学及乳房微循环的影响[J].中华中医药学刊,2008,(03):458-460.
[30]李振波，丘和明.活血化瘀法治疗癌症的讨论[J].中国中西医结合杂志,1996,16(9):559-561.
[31]韩俊庆，陈延条，满运艳，等.复春片合并放射治疗鼻咽癌临床研究[J].中国中西医结合杂志,1995,15(12):710-712.
[32]张凤瑞.膈下逐瘀汤现代研究概述[J].中成药,1995,17(5):44-45.
[33]李学汤.几种活血化瘀药物对小鼠肝癌细胞形成肺转移影响的初步试验观察[J].中医杂志,1980,21(8):78.
[34]黄孔威.赤芍对实验肿瘤生长和转移的影响及药理作用的研究[J].中华肿瘤杂志,1983,5(1):24.