川芎嗪联合CIK细胞对裸鼠肝癌HepG2细胞的影响
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摘要
目的探讨川芎嗪联合细胞因子诱导的杀伤(CIK)细胞对裸鼠肝癌HepG2细胞的影响。方法取HepG2细胞接种于裸鼠皮下,以建立裸鼠肝癌模型。建模成功后,按随机数字表法分为川芎嗪50 mg/L联合CIK细胞组(A组)、川芎嗪100 mg/L联合CIK细胞组(B组)和川芎嗪200 mg/L联合CIK细胞组(C组),各12只,灌胃给药1次;经尾静脉一次性输注1.5×10~7个CIK细胞。另将10只建模成功的裸鼠设为模型组(D组)。检测细胞侵袭和迁移能力,检测基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶抑制剂-2(TIMP-2)蛋白水平,检测细胞骨架蛋白-肌动蛋白微丝面积。结果与D组比较,B组和C组裸鼠迁移和侵袭细胞显著减少,抑制率显著升高(P <0. 05); A组、B组、C组裸鼠细胞MMP-2蛋白表达水平显著降低,TIMP-2蛋白表达水平显著升高(P <0. 05),细胞骨架蛋白-肌动蛋白微丝面积显著缩小(P <0. 05)。结论川芎嗪联合CIK细胞能抑制裸鼠肝癌HepG2细胞的迁移及侵袭,作用机制可能与下调细胞MMP-2和上调TIMP-2蛋白表达,以及抑制骨架微丝重排有关。
Objective To investigate the effect of ligustrazine combined with cytokine-induced killer( CIK) cells on hepatocellular carcinoma cell line HepG2 in nude mice. Methods HepG2 cells were inoculated subcutaneously in nude mice to establish models of hepatocellular carcinoma in nude mice. After the successful establishment of the models,the mice were divided into 50 mg/L ligustrazine combined with CIK cells group( group A),100 mg/L ligustrazine combined with CIK cells group( group B) and 200 mg/L ligustrazine combined with CIK cells group( group C) by the random number table method,12 mice in each group,all of them were given intragastric administration once,and 1. 5 × 10~7 CIK cells were transfused via tail vein at one time. Another 10 successfully modeled nude mice were selected as the model group( group D). The activity of cells' invasion and migration,the levels of matrix metalloproteinase-2( MMP-2),the tissue inhibitor of metalloproteinase-2( TIMP-2) protein and the area of cytoskeleton protein-actin microfilament were detected. Results Compared with group D,the migration cells and invasion cells of nude mice in group B and group C reduced significantly,while the inhibition rate in group B and group C increased significantly( P < 0. 05),the expression levels of MMP-2 protein in nude mice in group A,group B and group C decreased significantly,while the expression levels of TIMP-2 protein in group A,group B and group C increased significantly( P < 0. 05),the area of cytoskeleton protein-actin microfilament in group A,group B and group C reduced significantly( P < 0. 05). Conclusion Ligustrazine combined with CIK cells can inhibit the migration and invasion of HepG2 cells in nude mice. The mechanism may be related to down-regulation of MMP-2 protein expression and up-regulation of TIMP-2 protein expression,as well as inhibition of skeletal microfilament rearrangement.
Objective To investigate the effect of ligustrazine combined with cytokine-induced killer( CIK) cells on hepatocellular carcinoma cell line HepG2 in nude mice. Methods HepG2 cells were inoculated subcutaneously in nude mice to establish models of hepatocellular carcinoma in nude mice. After the successful establishment of the models,the mice were divided into 50 mg/L ligustrazine combined with CIK cells group( group A),100 mg/L ligustrazine combined with CIK cells group( group B) and 200 mg/L ligustrazine combined with CIK cells group( group C) by the random number table method,12 mice in each group,all of them were given intragastric administration once,and 1. 5 × 10~7 CIK cells were transfused via tail vein at one time. Another 10 successfully modeled nude mice were selected as the model group( group D). The activity of cells' invasion and migration,the levels of matrix metalloproteinase-2( MMP-2),the tissue inhibitor of metalloproteinase-2( TIMP-2) protein and the area of cytoskeleton protein-actin microfilament were detected. Results Compared with group D,the migration cells and invasion cells of nude mice in group B and group C reduced significantly,while the inhibition rate in group B and group C increased significantly( P < 0. 05),the expression levels of MMP-2 protein in nude mice in group A,group B and group C decreased significantly,while the expression levels of TIMP-2 protein in group A,group B and group C increased significantly( P < 0. 05),the area of cytoskeleton protein-actin microfilament in group A,group B and group C reduced significantly( P < 0. 05). Conclusion Ligustrazine combined with CIK cells can inhibit the migration and invasion of HepG2 cells in nude mice. The mechanism may be related to down-regulation of MMP-2 protein expression and up-regulation of TIMP-2 protein expression,as well as inhibition of skeletal microfilament rearrangement.
引文
[1]毕蕾,颜晓静,杨烨,等.川芎嗪对肝癌HepG2细胞迁移、侵袭和细胞骨架的影响[J].中国药理学通报,2016,25(2):194-198.
[2]赵建夫,赵凤芝,陈文慧,等.CIK细胞联合贝伐单抗对肝癌HepG2细胞的体内外抗瘤活性[J].中国肿瘤生物治疗杂志,2017,24(4):404-410.
[3]袁雅红,郭兴荣,于莉,等.酸性环境抑制CIK细胞对肝癌HepG2细胞的杀伤活性[J].肿瘤防治研究,2016,43(5):331-334.
[4]赵新,丛文铭,谭璐,等.MMP-2和TIMP-2在肝癌中的表达及其意义[J].临床肿瘤学杂志,2001,6(1):9-12.
[5]李克跃,石承先,汤可立,等.表柔比星联合川芎嗪对人肝癌细胞TGF-β1/Smads通路的影响[J].贵阳中医学院学报,2017,39(6):17-22.
[6]韩林涛,黄芳,胡爱萍,等.牛黄参干预裸鼠肝癌HepG2细胞移植瘤模型的代谢组学研究[EB/OL].(2017-04-17)[2018-08-16].http://www.paper.edu.cn/releasepaper/content/201704-176.
[7]左新华,高萍.DC-CIK免疫疗法联合TACE、RFA治疗原发性肝癌的疗效评估[J].海南医学院学报,2016,22(4):385-388.
[8]杨洋,陈和平,王占科,等.胰岛素和川芎嗪联合治疗对严重烫伤内毒素血症大鼠血浆s TREM-1、s ICAM-1、MDA和SOD水平的影响[J].解放军医药杂志,2017,29(6):10-14.
[9]张长洪,李琛,王布,等.川芎嗪联合顺铂对Lewis小鼠肺腺癌移植瘤与血管生成的影响[J].中国临床药理学杂志,2018,34(6):690-692.
[10]孙雯雯,窦金霞,张琳,等.山药提取物联合DC-CIK细胞疗法对结肠癌HT29干细胞荷瘤裸鼠的体内抗肿瘤研究[J].中草药,2017,48(7):1362-1368.
[11]文彬,孙海涛,贺松其,等.鳖甲煎丸对HepG2裸鼠移植瘤的抑制作用及瘤体组织中β-catenin、Tbx3表达水平的影响[J].南方医科大学学报,2016,36(2):210-214.
[2]赵建夫,赵凤芝,陈文慧,等.CIK细胞联合贝伐单抗对肝癌HepG2细胞的体内外抗瘤活性[J].中国肿瘤生物治疗杂志,2017,24(4):404-410.
[3]袁雅红,郭兴荣,于莉,等.酸性环境抑制CIK细胞对肝癌HepG2细胞的杀伤活性[J].肿瘤防治研究,2016,43(5):331-334.
[4]赵新,丛文铭,谭璐,等.MMP-2和TIMP-2在肝癌中的表达及其意义[J].临床肿瘤学杂志,2001,6(1):9-12.
[5]李克跃,石承先,汤可立,等.表柔比星联合川芎嗪对人肝癌细胞TGF-β1/Smads通路的影响[J].贵阳中医学院学报,2017,39(6):17-22.
[6]韩林涛,黄芳,胡爱萍,等.牛黄参干预裸鼠肝癌HepG2细胞移植瘤模型的代谢组学研究[EB/OL].(2017-04-17)[2018-08-16].http://www.paper.edu.cn/releasepaper/content/201704-176.
[7]左新华,高萍.DC-CIK免疫疗法联合TACE、RFA治疗原发性肝癌的疗效评估[J].海南医学院学报,2016,22(4):385-388.
[8]杨洋,陈和平,王占科,等.胰岛素和川芎嗪联合治疗对严重烫伤内毒素血症大鼠血浆s TREM-1、s ICAM-1、MDA和SOD水平的影响[J].解放军医药杂志,2017,29(6):10-14.
[9]张长洪,李琛,王布,等.川芎嗪联合顺铂对Lewis小鼠肺腺癌移植瘤与血管生成的影响[J].中国临床药理学杂志,2018,34(6):690-692.
[10]孙雯雯,窦金霞,张琳,等.山药提取物联合DC-CIK细胞疗法对结肠癌HT29干细胞荷瘤裸鼠的体内抗肿瘤研究[J].中草药,2017,48(7):1362-1368.
[11]文彬,孙海涛,贺松其,等.鳖甲煎丸对HepG2裸鼠移植瘤的抑制作用及瘤体组织中β-catenin、Tbx3表达水平的影响[J].南方医科大学学报,2016,36(2):210-214.