科罗拉多大学 苦瓜汁激活细胞能量传感器AMPK 导致人胰腺癌细胞凋亡

Bitter melon juice activates cellular energy sensor AMP-activated protein kinase causing apoptotic death of human pancreatic carcinoma cells

　

　

Carcinogenesis. 2013 Jul; 34(7): 1585–1592  

By Colorado University

Manjinder Kaur, 1 Gagan Deep, 1,2 Anil K. Jain, 1 Komal Raina, 1 Chapla Agarwal, 1, 2 Michael F. Wempe, 1, 2和Rajesh Agarwal 1, 2，*

 

胰腺癌的预后非常差，这表明需要额外的药物来改善疾病预后。

 

在这项研究中，我们检测了一种新型制剂苦瓜汁(BMJ)在培养和裸鼠中对胰腺癌细胞的作用及其相关机制。

 

采用3-(4,5-二甲基噻唑-2基)-2,5-二苯基四唑溴化钠、细胞死亡酶联免疫吸附试验和annexin/ propidide检测，分析了BMJ在人胰腺癌BxPC-3、MiaPaCa-2、AsPC-1和Capan-2细胞中的抗癌作用。

 

采用免疫印迹法检测BMJ对凋亡调节因子的影响。在体内BMJ治疗裸鼠MiaPaCa-2肿瘤的疗效进行评估，并采用免疫组织化学(IHC)分析异种移植的生物标志物。结果显示，BMJ (2-5% v/v)通过诱导细胞凋亡而降低所有4个胰腺癌细胞系的细胞活力。

　

 

在分子水平上，BMJ引起caspases活化，Bcl-2家族成员表达改变，细胞色素C释放到胞质中。

 

此外，BMJ降低了survivin和x -连锁抑制凋亡蛋白的表达，但增加了p21、CHOP和磷酸化的丝裂原活化蛋白激酶(细胞外信号调节激酶1/2和p38)水平。

 

重要的是，BMJ激活腺苷单磷酸酯活化蛋白激酶(AMPK)是细胞能量状态的生物标志物，AMPK抑制剂(化合物C)逆转了BMJ诱导的caspase-3激活，提示激活AMPK参与BMJ诱导的凋亡。

　

 

体内,口服冻干BMJ(5毫克/100µl水/天/鼠)为6周抑制MiaPaCa-2肿瘤异种移植增长了60%(P < 0.01),没有明显的裸体小鼠的毒性。

 

IHC对MiaPaCa-2异种移植体的分析表明，BMJ在体内也能抑制增殖、诱导凋亡和激活AMPK。

 

总的来说，BMJ在体外和体内对人类胰腺癌细胞都有很强的抗癌作用，表明其临床应用价值。

 

Bitter melon juice activates cellular energy sensor AMP-activated protein kinase causing apoptotic death of human pancreatic carcinoma cells

 

COLORADO UNIVERSITY

Manjinder Kaur, 1 Gagan Deep, 1 , 2 Anil K. Jain, 1 Komal Raina, 1 Chapla Agarwal, 1 , 2 Michael F. Wempe, 1 , 2 and Rajesh Agarwal 1 , 2 ,*

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Abstract

 

Prognosis of pancreatic cancer is extremely poor, suggesting critical needs for additional drugs to improve disease outcome. In this study, we examined efficacy and associated mechanism of a novel agent bitter melon juice (BMJ) against pancreatic carcinoma cells both in culture and nude mice. BMJ anticancer efficacy was analyzed in human pancreatic carcinoma BxPC-3, MiaPaCa-2, AsPC-1 and Capan-2 cells by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide, cell death enzyme-linked immunosorbent assay and annexin/propidium iodide assays. BMJ effect on apoptosis regulators was assessed by immunoblotting. In vivo BMJ efficacy was evaluated against MiaPaCa-2 tumors in nude mice, and xenograft was analyzed for biomarkers by immunohistochemistry (IHC). Results showed that BMJ (2–5% v/v) decreases cell viability in all four pancreatic carcinoma cell lines by inducing strong apoptotic death. At molecular level, BMJ caused caspases activation, altered expression of Bcl-2 family members and cytochrome-c release into the cytosol. Additionally, BMJ decreased survivin and X-linked inhibitor of apoptosis protein but increased p21, CHOP and phosphorylated mitogen-activated protein kinases (extracellular signal-regulated kinase 1/2 and p38) levels. Importantly, BMJ activated adenosine monophosphate-activated protein kinase (AMPK), a biomarker for cellular energy status, and an AMPK inhibitor (Compound C) reversed BMJ-induced caspase-3 activation suggesting activated AMPK involvement in BMJ-induced apoptosis. In vivo, oral administration of lyophilized BMJ (5mg in 100 µl water/day/mouse) for 6 weeks inhibited MiaPaCa-2 tumor xenograft growth by 60% (P < 0.01) without noticeable toxicity in nude mice. IHC analyses of MiaPaCa-2 xenografts showed that BMJ also inhibits proliferation, induces apoptosis and activates AMPK in vivo. Overall, BMJ exerts strong anticancer efficacy against human pancreatic carcinoma cells, both in vitro and in vivo, suggesting its clinical usefulness.

 

http://europepmc.org/articles/PMC3697895