松果菊苷对结肠癌SW480细胞体内外增殖、侵袭和转移的影响  

Effects of Echinacoside on Proliferation,Invasion and Metastasis of Colon Cancer SW480 Cells in Vitro and in Vivo

作　　者：韩一梅 靳伟民[1] 曾会[1] 王成[1] 张辉[1] HAN Yi-Mei;JIN Wei-Min;ZENG Hui;WANG Cheng;ZHANG Hui(Dept.of Gastroenterology,Hanzhong Central Hospital,Hanzhong 723000 Shaanxi,China)

机构地区：[1]汉中市中心医院消化内科,陕西汉中723000

出　　处：《广州中医药大学学报》2020年第8期1542-1549,共8页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：陕西省卫生计生科研基金资助项目(编号:2016E035)。

摘　　要：【目的】探讨松果菊苷对结肠癌细胞体内外增殖、侵袭和转移的影响及机制。【方法】(1)体外实验:以松果菊苷0(作为空白对照)、10、25、50μmol/L处理结肠癌SW480细胞24 h后,采用克隆形成实验、BrdU染色法测定细胞增殖能力;流式细胞术测定细胞凋亡率;Transwell实验测定细胞侵袭能力;划痕实验测定细胞迁移能力。(2)体内实验:将60只BALB/c裸鼠随机分为空白对照组,松果菊苷10、25、50 mg/kg组,每组15只。给予所有裸鼠背部皮下注射人结肠癌SW480细胞悬液构建结肠癌移植瘤模型。同时,松果菊苷10、25、50 mg/kg组裸鼠分别腹腔注射相应浓度的松果菊苷溶液,空白对照组裸鼠腹腔注射等量生理盐水,每天1次,连续注射28 d。记录裸鼠存活情况和移植瘤体积。给药结束后,免疫组织化学法测定移植瘤中Ki67、血管内皮细胞生长因子(VEGF)的表达水平;原位末端脱氧核苷酸转移酶(TdT)标记(TUNEL)法测定移植瘤中肿瘤细胞的凋亡水平;蛋白质免疫印迹(Western Blot)法测定移植瘤中E钙黏蛋白(E-cadherin)、N钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)的表达水平。【结果】与空白对照组比较,松果菊苷10、25、50μmol/L组细胞克隆数量、BrdU阳性细胞百分比均降低,细胞凋亡率均增加,侵袭细胞数目和划痕闭合率均降低,呈剂量依赖性,松果菊苷25、50μmol/L组差异有统计学意义(P <0.05或P <0.01)。与空白对照组比较,松果菊苷10、25、50 mg/kg组裸鼠存活率均增加,移植瘤体积均减小,移植瘤细胞凋亡率均增加,移植瘤中VEGF、Ki67、N-cadherin、Vimentin表达水平均降低,E-cadherin、cl-caspase-3和cl-caspase-9表达水平均升高(P <0.05或P <0.01),呈剂量依赖性,松果菊苷25、50 mg/kg组差异有统计学意义(P <0.05或P <0.01)。【结论】松果菊苷可抑制人结肠癌细胞体内外增殖、侵袭和转移,促进移植瘤裸鼠存活,其机制与促进结肠癌细胞凋亡,调控上�Objective To explore the effects of echinacoside on proliferation, invasion and metastasis of coloncancer cells in vitro and in vivo and its mechanism. Methods(1)In Vitro:After colon cancer SW480 cells weredealed with echinacoside of 0(as blank control), 10, 25, 50 μmol/L for 24 h, the cell proliferation wasexamined by colony formation assay and Brd U staining method,cell apoptosis by flow cytometry,cell invasion byTranswell assay,and cell migration by scratch assay.(2)In Vivo:Sixty nude mice were randomly divided intoblank control group and echinacoside 10,25,50 mg/kg groups,15 nude mice in each group. All the nude micewere induced into transplanted tumor model by dorsal subcutaneous injection of SW480 cell suspension.Meanwhile, the nude mice in echinacoside 10, 25, 50 mg/kg groups were given intraperitoneal injection ofcorresponding concentrations of echinacoside,and the blank control group was given intraperitoneal injection ofthe same volume of normal saline,once per day,for 28 continuous days. The survival and transplanted tumorvolume were recorded. After medication the expression levels of Ki67 and endothelial growth factor(VEGF)in transplanted tumor were detected by immunohistochemistry, tumor cell apoptosis in transplanted tumor wasdetermined by terminal deoxyribonucleotidyl transferse(Td T)-mediated biotin-16-d UTP nick-end labelling(TUNEL) assay, the expression levels of E-cadherin, N-cadherin, Vimentin in transplanted tumor weredetected by Western blotting method. Results Compared with the blank control group,the number of clones andthe proportion of Brd U-positive cells were decreased, the cell apoptosis rate was increased, the number ofinvasive cells and cell migration rate were decreased in the echinacoside 10, 25, 50 mg/kg groups in dose-dependent manner,the difference of the echinacoside 25,50 mg/kg groups being significant(P < 0.05 or P <0.01). Compared with the blank control group,the survival rate of mice was increased,the volume of tumors wasdecreased,the cell apoptosis rate of transplanted t

关 键 词：松果菊苷 结肠癌 细胞增殖 细胞侵袭 细胞转移 裸鼠 

分 类 号：R285.5[医药卫生—中药学]