雷公藤甲素对裸鼠胰腺癌皮下移植瘤生长的抑制作用及其机制
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摘要
目的:观察雷公藤甲素对人胰腺癌荷瘤裸鼠的抑瘤作用,探讨其作用的分子机制。方法:选择健康BALB/c裸鼠于右后肢皮肤移植人胰腺癌SW1990细胞,建立裸鼠荷瘤模型,将建模成功的裸鼠分为模型组、吉西他滨组和雷公藤甲素组,每组10只,另选10只健康裸鼠作为对照组。测量各组SW1990胰腺癌荷瘤裸鼠体质量,取肿瘤组织称质量,计算肿瘤抑制率;采用免疫组织化学法检测各组裸鼠胰腺组织中凋亡相关蛋白Bax和Bcl-2的表达情况;采用Western blotting法检测各组裸鼠胰腺组织中凋亡相关蛋白Bax、Bcl-2、Caspase-9和Caspase-3表达水平。结果:与吉西他滨组比较,雷公藤甲素组SW1990胰腺癌的肿瘤抑制率明显升高(P<0.05)。HE染色观察,与模型组比较,雷公藤甲素组裸鼠胰腺癌肿瘤细胞增殖受到抑制。免疫组织化学检测,与模型组比较,雷公藤甲素组裸鼠胰腺组织中Bcl-2表达水平和Bcl-2/Bax比值明显降低(P<0.05)。Western blotting法检测,与模型组比较,雷公藤甲素组裸鼠胰腺组织中Bax、Caspase-9和Caspase-3蛋白表达水平均明显升高(P<0.05),而Bcl-2蛋白表达水平明显降低(P<0.05)。结论:雷公藤甲素能抑制裸鼠肿瘤组织的生长,其机制可能与抑制胰腺肿瘤组织中Bcl-2的激活、促进Bax的表达、降低Bcl-2/Bax的比值、促使Caspase-9和Caspase-3激活及诱导胰腺癌移植瘤细胞凋亡有关。
Objective:To observe the antitumor effect of triptolide in the human pancreatic cancer tumor-bearing nude mice,and to explore its molecular mechanism.Methods:The healthy BALB/c nude mice were selected and transplanted with the human pancreatic cancer SW1990 cells into the skin of the right hind limb to establish the tumor-bearing model of nude mice.The successful modeling nude mice were divided into model group,gemcitabine group and triptolide group,10 in each group;another 10 healthy nude mice were selected as control group.The tumor tissue of SW1990 pancreatic cancer tumor-bearing nude mice were weighed and the inhibitory rate of tumor was calculated.The expressions of apoptosis-related proteins Bax and bcl-2 in the pancreas tissue of the nude mice in each group were detected by immunohistochemistry.The expression levels of Bax,bcl-2,Caspase-9 and Caspase-3 in the pancreas tissue of the nude mice in each group were detected by Western blotting method.Results:Compared with gemcitabine group,the inhibitory rate of tumor of SW1990 pancreatic cancer in triptolide group was significantly increased(P <0.05).The HE staining results showed that compared with model group,the proliferation of pancreatic cancer tumor cells in triptolide group was inhibited.The immunohistochemistry results showed that the expression level of Bcl-2 in the pancreas tissue and the Bcl-2/Bax ratio of the nude mice in tripptolide group were lower than those in model group(P<0.05).The Western blotting results showed that compared with model group,the expression levels of Bax,Caspase-9 and Caspase-3 in the pancreas tissue of the nude mice in triptolide group were significantly increased(P<0.05),while the expression level of Bcl-2 was significantly decreased(P<0.05).Conclusion:Triptolide can inhibit the growth of tumor tissue in the nude mice;its mechanism may be related to inhibiting the activation of Bcl-2 in pancreatic tumor tissue,promoting the expression of Bax,reducing the ratio of Bcl-2/Bax,promoting the activation of Caspase-9 and Caspase-3,and inducing the apoptosis of pancreatic cancer transplanted tumor cells.
Objective:To observe the antitumor effect of triptolide in the human pancreatic cancer tumor-bearing nude mice,and to explore its molecular mechanism.Methods:The healthy BALB/c nude mice were selected and transplanted with the human pancreatic cancer SW1990 cells into the skin of the right hind limb to establish the tumor-bearing model of nude mice.The successful modeling nude mice were divided into model group,gemcitabine group and triptolide group,10 in each group;another 10 healthy nude mice were selected as control group.The tumor tissue of SW1990 pancreatic cancer tumor-bearing nude mice were weighed and the inhibitory rate of tumor was calculated.The expressions of apoptosis-related proteins Bax and bcl-2 in the pancreas tissue of the nude mice in each group were detected by immunohistochemistry.The expression levels of Bax,bcl-2,Caspase-9 and Caspase-3 in the pancreas tissue of the nude mice in each group were detected by Western blotting method.Results:Compared with gemcitabine group,the inhibitory rate of tumor of SW1990 pancreatic cancer in triptolide group was significantly increased(P <0.05).The HE staining results showed that compared with model group,the proliferation of pancreatic cancer tumor cells in triptolide group was inhibited.The immunohistochemistry results showed that the expression level of Bcl-2 in the pancreas tissue and the Bcl-2/Bax ratio of the nude mice in tripptolide group were lower than those in model group(P<0.05).The Western blotting results showed that compared with model group,the expression levels of Bax,Caspase-9 and Caspase-3 in the pancreas tissue of the nude mice in triptolide group were significantly increased(P<0.05),while the expression level of Bcl-2 was significantly decreased(P<0.05).Conclusion:Triptolide can inhibit the growth of tumor tissue in the nude mice;its mechanism may be related to inhibiting the activation of Bcl-2 in pancreatic tumor tissue,promoting the expression of Bax,reducing the ratio of Bcl-2/Bax,promoting the activation of Caspase-9 and Caspase-3,and inducing the apoptosis of pancreatic cancer transplanted tumor cells.
引文
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[2]武赞凯,杜恒锐,王振江,等.胰腺癌流行病学及诊治的研究进展[J].中南大学学报:医学版,2017,42(6):713-719.
[3]SHEN Y F,ZHANG X,WANG Y,et al.Celastrol targetsIRKs to block Toll-like 4-mediated nuclear factor-κBactivation[J].J Integr Med,2016,14(3):203-208.
[4]乔志新,贺敏,李伟静,等.雷公藤甲素诱导胰腺癌细胞凋亡[J].生物技术通讯,2013,24(4):519-523.
[5]郑婷婷,焦丽静,阙祖俊,等.中药诱导肿瘤凋亡相关机制研究进展[J].上海中医药杂志,2016,50(10):103-109.
[6]ZHANG Y,LI Z,BA W Q,et al.Dermal pharmacokinetic study of Fufang Leigongteng microemulsion gel by microdialysis combined with HPLC[J].Pharm Today,2015,25(9):620-623.
[7]ZIAEI S,HAIABY R.Immunosuppressive,anti-inflammatory and anti-cancer properties of triptolide:Aminireview[J].Avicenna J Phytomed,2016,6(2):149-164.
[8]WEI Y M,WANG Y H,XUE H Q,et al.Triptolide,a potential autophagy modulator[J].Chin J Integr Med,2019,25(3):233-240.
[9]VAZQUEZ-SANTILLAN K,MELENDEZ-ZAJGLA J,JIMENEZ-HERNANDEZ L,et al.NF-κB signaling in cancer stem cells:apromising therapeutic target?[J].Cell Oncol(Dordr),2015,38(5):327-339.
[10]XI C,PENG S J,WU Z P,et al.Toxicity of triptolide and the molecular mechanisms involved[J].Biomed Pharmacother,2017,90(3):531-541.
[11]张艳,李海龙,王虎平,等.阿魏酸对人胃癌MGC-803细胞增殖的影响[J].中国中医药信息杂志,2016,23(9):70-73.
[12]陈豫民,王琰,韩轲,等.bcl-2结合抗凋亡蛋白在食管癌中的表达及意义[J].山西医药杂志,2015,44(16):1873-1875.
[13]孙云,胡晓杰,马国娟,等.胃癌组织中LMO1与Bcl-2、Bax、Survivin、Caspase-3蛋白表达的关系及意义[J].肿瘤学杂志,2016,22(9):703-707.
[14]曹米兰,吴有芳,郭建军.不同方法处理的九香虫对人胃癌SGC-7901细胞体外作用比较及其抑癌组分分布[J].生物资源,2017,39(5):328-332.
[15]杨静,凋亡相关基因Caspase-3、Caspase-9和c-IAP1在胃腺癌组织中的表达及意义[D].承德:承德医学院,2017.
[16]高德全,张建波,王联林,等.胃癌组织EZH2与凋亡相关蛋白Caspase-3、Bcl-2、Bax表达的关系及意义[J].现代中西医结合杂志,2014,23(15):1615-1617.
[17]张妍,何航,韩倩倩,等.α-细辛醚对Eca-109细胞凋亡及XIAP、caspase-3表达的影响[J].郑州大学学报:医学版,2017,52(4):377-380.
[18]YU X L,ZHOU X,FU C L,et al.Celastrol induces apoptosis of human osteosarcoma cells via the mitochondrialapoptotic pathway[J].Oncol Rep,2015,34(3):1129-1136.
[19]CASC O R,FONSECA J E,MOITA L F.Celastrol:Aspectrum of treatment opportunities in chronic diseases[J].Front Med(Lausanne),2017,15(4):69.
[20]LI H Y,ZHANG J,SUN L L,et al.Celastrol induces apoptosis and autophagy via the ROS/JNK signaling pathway in human osteosarcoma cells:An in vitro and in vivo study[J].Cell Death Dis,2015,22(6):e1604.