蝙蝠葛碱抑制人胰腺癌SW1900细胞系增殖及诱导其凋亡

张珦

doi:10.16098/j.issn.0529-1356.2020.04.011

PDF(1490 KB)

解剖学报 ›› 2020, Vol. 51 ›› Issue (4) : 543-547. DOI: 10.16098/j.issn.0529-1356.2020.04.011

肿瘤生物学

蝙蝠葛碱抑制人胰腺癌SW1900细胞系增殖及诱导其凋亡

张珦¹ 范竞²章宜芬¹李惠¹李长印^3*

作者信息 +

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

ZHANG Xiang¹ FAN Jing² ZHANG Yi-fen¹ LI Hui¹LI Chang-yin^3*

Author information +

文章历史 +

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。

Abstract

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 Annexin Ⅴ-FITC/PI fluorescent staining of flow cytometric method . The expressions of phosphatidylinositol 3-kinase (PI3K), 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 PI3K, Akt and Bcl-2 were lower obviously (PI3K 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 PI3K, 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 PI3K/Akt signal pathway and lower Bcl-2 expression.

导出引用

张珦范竞章宜芬李惠李长印. 蝙蝠葛碱抑制人胰腺癌SW1900细胞系增殖及诱导其凋亡[J]. 解剖学报. 2020, 51(4): 543-547 https://doi.org/10.16098/j.issn.0529-1356.2020.04.011

ZHANG Xiang FAN Jing ZHANG Yi-fen LI Hui LI Chang-yin. Dauricine inhibiting the cell proliferation and inducing the cell apoptosis of human pancreatic cancer cells line SW1900#br#[J]. Acta Anatomica Sinica. 2020, 51(4): 543-547 https://doi.org/10.16098/j.issn.0529-1356.2020.04.011

中图分类号： R735.9

参考文献

［1］ Liu LM. Opportunities and challenges of integrated traditional Chinese and western medicine in the treatment of pancreatic cancer［J］. Chinese Journal of Integrated Traditional and Western Medicine, 2019, 39(1):18-20. (in Chinese)

刘鲁明. 中西医结合治疗胰腺癌的机遇与挑战［J］. 中国中西医结合杂志, 2019, 39(1):18-20.

［2］ Zhang YB, Zhou ZhG, Zhong LL, et al. Research progress on antitumor activities of dauricine［J］. China Medical Equipment,2014,11(12):370. (in Chinese)

张英博,周忠光,仲丽丽,等.蝙蝠葛碱抗肿瘤研究新进展［J］.中国医学装备,2014,11(12):3701.

［3］ Cui L, Pan YSh. Inhibitory effect of tetrandrine and dauricine on growth of HL-60 and K562 cell lines［J］. Chinese Pharmacological Bulletin,1995(6):478-481. (in Chinese)

崔燎,潘毅生.粉防己碱和蝙蝠葛碱对人白血病细胞株HL-60和K562的生长抑制作用［J］.中国药理学通报,1995(6):478-481.

［4］ Yang WSh, Sun Sh, Lü H. Detection of anti-tumor proliferation of active components of the menispermum dauricum by MTT method［J］. Journal of Medical Science Yanbian University,2010,33(4):261-262. (in Chinese)

杨万山,孙抒,吕贺.MTT法检测蝙蝠葛活性成分的抑制肿瘤增殖作用［J］.延边大学医学学报,2010,33(4):261-262.

［5］ Yang Z, Li C, Wang X, et al. Dauricine induces apoptosis, inhibits proliferation and invasion through inhibiting NF-κB signaling pathway in colon cancer cells［J］. J Cell Physiol,2010,225(1):266-275.

［6］ Zhang ShB, Li HB. Mechanism research on the proliferation inhibition of human breast cancer cells MCF-7 by dauricin with PTEN/PI3K/Akt pathway［J］. Journal of Chinese Medicinal Materials,2018,41(10):2181-2184.(in Chinese)

张术波,李海滨.蝙蝠葛碱通过调控PTEN/PI3K/Akt通路抑制人乳腺癌MCF-7细胞增殖与诱导凋亡［J］.中药材,2018,41(10):2181-2184.

［7］ Li M, Liang WJ, Zeng YP, et al. Inhibitory effect of dauricine on growth of human bladder cancer T-24 cell line［J］. Acta Academiae Medicinae Militaris Tertiae,2006,28(18):1860-1863.(in Chinese)

李铭,梁文杰,曾亚平,等.蝙蝠葛碱对人膀胱癌T-24细胞株生长增殖的抑制作用［J］.第三军医大学学报,2006,28(18):1860-1863.

［8］ Zhang S, Ren Y, Qiu J. Dauricine inhibits viability and induces cell cycle arrest and apoptosis via inhibiting the PI3K/Akt signaling pathway in renal cell carcinoma cells［J］. Mol Med Rep,2018,17(5): 7403-7408.

［9］ Wang J, Li Y, Zu XB, et al. Dauricine can inhibit the activity of proliferation of urinary tract tumor cells［J］. Asian Pac J Trop Med,2012, 5(12):973-976.

［10］ Ding H, Tang XD, Zhou KY. Effects of dauricine and piperine on proliferation of two nasopharyngeal cell lines CNE-1 and CNE-2Z［J］. Journal of Guangdong Medical College,2003,21(1):16-17.(in Chinese)

丁航,唐旭东,周克元.蝙蝠葛碱和胡椒碱对两株鼻咽癌细胞增殖的影响［J］.广东医学院学报,2003,21(1):16-17.

［11］ Schmittgen TD, Livak KJ. Analyzing Real-time PCR data by the comparative C(T) method［J］. Nat Protc, 2008,3(6):1101-1108.

［12］ Chen W, Wang SX, Hu XM,et al. Atomic force microscope-related study on dauricine induced apoptosis of NB4 cell［J］. Journal of Sun Yat-sen University(Medical Sciences),2012,33(4):421-428.(in Chinese)

陈伟,王穗湘,胡小毛,等.基于AFM对蝙蝠葛碱诱导NB4细胞凋亡的研究［J］.中山大学学报(医学科学版),2012,33(4):421-428.

［13］ Shi YSh, Zhang Y, Wang JZh, et al. Inhibitory effect of dauricine on growth of human lung cancer QG-56 cell line［J］. Information on Traditional Chinese Medicine,2010,27(3):115-116.(in Chinese)

石玉生,张艳,王加志,等.蝙蝠葛碱抑制肺癌细胞QG-56增殖的实验研究［J］.中医药信息,2010,27(3):115-116.

［14］ Xie WR, Zhang G. Research progress of PI3K/AKT/Bcl-2 apoptosis signal transduction pathway［J］. China Modern Medicine,2015,22(30):22-25.(in Chinese)

谢玮蓉,张刚.PI3K/AKT/Bcl-2凋亡信号传导通路的研究进展［J］.中国当代医药,2015,22(30):22-25.

［15］ Zou ZhJ, Gao ZG, Wang XM, et al. Effect of curcumin on PI3K/Akt/Bcl-2 pathway signaling lung adenocarcinoma cell A549［J］. Journal of Jiangxi University of Traditional Chinese Medicine,2013,25(5):15-17.(in Chinese)

邹志坚,高增光,王晓敏,等.姜黄素对肺腺癌A549细胞PI3K/Akt/Bcl-2信号通路的作用［J］.江西中医学院学报,2013,25(5):15-17.

［16］ Yao X, Lin JY, Guo D, et al. Mechanism research on the proliferation inhibition of human prostate cancer cells LNCaP by Tengcha extract with PI3K/Akt/Bcl-2 pathway［J］. Fujian Medical Journal,2018,40(2):105-107.(in Chinese)

姚欣,林静瑜,郭丹,等.藤茶提取物通过PI3K/Akt/Bcl-2途径抑制人前列腺癌LNCaP细胞增殖的机制研究［J］.福建医药杂志,2018,40(2):105-107.

［17］ Na M, Guo RZh, Ruan Y, et al. Correlation investigate PI3K-AKT-Bcl-2 anti-apoptotic pathway with scar carcinoma cells apoptosis［J］. Chinese Journal of Cancer Prevention and Treatment,2014,21(8):599-604.

马娜,郭瑞珍,阮媛,等.PI3K-AKT-Bcl-2抗凋亡途径与瘢痕癌癌细胞凋亡相关性探讨［J］.中华肿瘤防治杂志,2014,21(8):599-604.