蝙蝠葛碱抑制人胰腺癌SW1900细胞系增殖及诱导其凋亡  

Dauricine inhibiting the cell proliferation and inducing the cell apoptosis of human pancreatic cancer cells line SW1900

作　　者：张珦[1] 范竞 章宜芬[1] 李惠[1] 李长印[3] ZHANG Xiang;FAN Jing;ZHANG Yi-fen;LI Hui;LI Chang-yin(Department of Pathology,Jiangsu Province Hospit of Chinse Medicine,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China;Department of Clinical Education,AOMA Graduate School of Integrative Medicine,Texas 78745,USA;Clinical Pharmacology Laboratory,Jiangsu Province Hospital of Chinese Medicine,Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China)

机构地区：[1]江苏省中医院南京中医药大学附属医院病理科,南京210029 [2]奥斯汀综合医学研究生院临床教学部,美国德克萨斯州78745 [3]江苏省中医院南京中医药大学附属医院临床药理实验室,南京210029

出　　处：《解剖学报》2020年第4期543-547,共5页Acta Anatomica Sinica

基　　金：国家自然科学青年基金(81503300);江苏省中医院高峰人才项目(y2018rc22)。

摘　　要：目的探讨蝙蝠葛碱对人胰腺癌SW1900细胞增殖与凋亡的影响及其机制。方法用不同浓度蝙蝠葛碱处理细胞系SW1900,采用MTT法检测细胞增殖,采用流式细胞术Annexin V-FITC/PI双染法测定细胞凋亡率。Real-time PCR和免疫印迹法分析蝙蝠葛碱处理SW1900细胞中凋亡蛋白及磷脂酰肌醇-3激酶/蛋白激酶B(PI3K/Akt)的表达水平。结果MTT实验显示,蝙蝠葛碱呈剂量依赖性地抑制SW1900细胞活力,F=783.7,P<0.001。流式细胞术结果显示,经0、6、12 mg/L蝙蝠葛碱处理的各组细胞凋亡率分别为(4.34±1.30)%、(14.94±1.94)%和(22.68±3.61)%,3组间均值差异有统计学意义,F=58.52,P<0.001,蝙蝠葛碱呈剂量依赖性促进细胞凋亡。Real-time PCR实验结果显示,蝙蝠葛碱可剂量依赖性下调PI3K、Akt、Bcl-2的基因表达(PI3K mRNA,F=101,P=0.01;Akt mRNA,F=1666,P<0.01;Bcl-2 mRNA,F=753,P<0.001)。免疫印迹法结果显示,蝙蝠葛碱呈剂量依赖性地下调PI3K、Akt和Bcl-2的蛋白表达。结论蝙蝠葛碱对人胰腺癌SW1900细胞具有抑制其增殖和诱导其凋亡等作用,可能通过PI3K/Akt通路下调Bcl-2的表达诱导SW1900细胞凋亡。Objective To discuss the proliferation inhibition and apoptosis induction of human pancreatic cancer cell line SW1900 by dauricine and its possible mechanism.Methods The MTT colorimetric method was used to detect the inhibitory effects of cell viability.The apoptosis rate was tested by the AnnexinⅤ-FITC/PI fluorescent staining of flow cytometric method.The expressions of phosphatidylinositol 3-kinase(PI3 K),protein kinase B(Akt)and B-cell lymphoma-2(Bcl-2)were detected by Real-time PCR and Western blotting assay.Results MTT assay showed that dauricine significantly inhibited the proliferation of SW1900 cells and the inhibitory effect was enhanced with the increasing of dauricine concentration,F=783.7,P<0.001.The apoptosis of 3 groups cells were(4.34±1.30)%(0 mg/L dauricine),(14.94±1.94)%(6 mg/L dauricine)and(22.68±3.61)%(12 mg/L dauricine).The mean difference was statistically significant among the three groups(F=58.52,P<0.001).Dauricine could significantly induce apoptosis human pancreatic cancer cells with dose-dependent manner.Real-time PCR showed that the gene expressions of PI3 K,Akt and Bcl-2 were lower obviously(PI3 K mRNA,F=101,P=0.01;Akt mRNA,F=1666,P<0.01;Bcl-2 mRNA,F=753,P<0.001)with dose-dependent manner.Western blotting assay also showed that the protein expression of PI3 K,Akt and Bcl-2 was down-regulated with dose-dependent manner.Conclusion Dauricine has proliferation inhibition and apoptosis inducement effect on human pancreatic cancer cells line SW1900.This function may be concerned with the regulation of PI3 K/Akt signal pathway and lower Bcl-2 expression.

关 键 词：蝙蝠葛碱 胰腺癌 SW1900细胞 磷脂酰肌醇-3激酶 蛋白激酶B BCL-2基因 免疫印迹法 人 

分 类 号：R735.9[医药卫生—肿瘤]