Full Text:   <905>

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CLC number: R96

On-line Access: 2017-06-05

Received: 2016-08-23

Revision Accepted: 2016-12-04

Crosschecked: 2017-05-10

Cited: 0

Clicked: 1759

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Lan-lan Wan

http://orcid.org/0000-0002-3728-2051

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Journal of Zhejiang University SCIENCE B 2017 Vol.18 No.6 P.522-531

http://doi.org/10.1631/jzus.B1600369


Anti-hepatocarcinoma activity of TT-1, an analog of melittin, combined with interferon-α via promoting the interaction of NKG2D and MICA


Author(s):  Lan-lan Wan, Da-qi Zhang, Jin-nan Zhang, Li-qun Ren

Affiliation(s):  Department of Experimental Pharmacology and Toxicology, School of Pharmaceutical Sciences, Jilin University, Changchun 130021, China; more

Corresponding email(s):   renlq_ren@sina.com

Key Words:  TT-1, Interferon-α, (IFN-α, ), Natural killer (NK) cells, Hepatocarcinoma, Immunotherapy


Lan-lan Wan, Da-qi Zhang, Jin-nan Zhang, Li-qun Ren. Anti-hepatocarcinoma activity of TT-1, an analog of melittin, combined with interferon-α via promoting the interaction of NKG2D and MICA[J]. Journal of Zhejiang University Science B, 2017, 18(6): 522-531.

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author="Lan-lan Wan, Da-qi Zhang, Jin-nan Zhang, Li-qun Ren",
journal="Journal of Zhejiang University Science B",
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publisher="Zhejiang University Press & Springer",
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%T Anti-hepatocarcinoma activity of TT-1, an analog of melittin, combined with interferon-α via promoting the interaction of NKG2D and MICA
%A Lan-lan Wan
%A Da-qi Zhang
%A Jin-nan Zhang
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A1 - Lan-lan Wan
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Abstract: 
hepatocarcinoma is one of the malignant cancers with significant morbidity and mortality. immunotherapy has emerged in clinical treatment, owing to the limitation and severe side effects of chemotherapy. In the immune system, natural killer (NK) cells are important effectors required to eliminate malignant tumor cells without the limitation of major histocompatibility complex (MHC) molecule issues. Hence, treatment which could stimulate NK cells is of great interest. Here, we investigated the efficacy of the combined therapy of TT-1 (a mutant of melittin) and interferon-α; (IFN-α;) on NK cells and human liver cancer HepG-2/Huh7 cells in vitro and in vivo, as well as the mechanism involved. The combination therapy significantly inhibited the growth of HepG-2/Huh7 cells in vivo, but this effect was impaired after depleting NK cells. TT-1 not only up-regulated MHC class I-related chain molecules A (MICA) expression, but also prevented the secretion of soluble MICA (sMICA). Both the mRNA and protein of a disintegrin and metallopeptidase 10 (ADAM 10) in HepG-2/Huh7 cells were decreased after TT-1 treatment. The combined therapy of TT-1 and IFN-α could suppress the growth of HepG-2/Huh7 xenografted tumor effectively via promoting the interaction of NK group 2, member D (NKG2D) and MICA, indicating that TT-1+IFN-α would be a potential approach in treating liver cancer.

联合使用IFN-α和TT-1通过增强NKG2D和MICA的相互作用达到抗肝癌效果的研究

目的:评估干扰素α(IFN-α)和TT-1(一种蜂毒肽的类似物)联合用药的抗肿瘤效果,并初步研究联合用药的抗肿瘤及免疫调节机制。
创新点:为了增强蜂毒肽的抗肿瘤效果,本课题组在其基础上进行改造,合成了一种新的化合物TT-1。该研究第一次将蜂毒肽类似物和免疫细胞因子IFN-α联合使用,并通过实验证实联合用药可以通过激活免疫调节来增强TT-1的抗肿瘤效果。
方法:首先通过MTT实验验证TT-1对HepG-2/Huh7细胞的增殖抑制作用。接着建立HepG-2/Huh7小鼠移植瘤模型,考察TT-1+IFN-α的体内抗肿瘤效果;使用anti-asialo GM-1抗体消除自然杀伤(NK)细胞,验证NK细胞在联合用药中的关键作用。使用流式细胞术和酶联免疫吸附法(ELISA)验证TT-1对HepG-2/Huh7细胞MHC I链相关分子A(MICA)表达的影响,并用实时聚合酶联反应(RT-PCR)和蛋白质印迹(Western blot)对其机制进行探究;通过细胞毒性实验考察TT-1+IFN-α是否可以增强NK细胞对HepG-2/ Huh7细胞的特异性杀伤作用。最后使用免疫组化的方法考察TT-1+IFN-α联合用药对肿瘤组织中MICA和NKG2D的表达量的影响。
结论:MTT实验表明TT-1可以在体外有效地抑制HepG-2/Huh7细胞的增殖。小鼠移植瘤模型实验结果显示TT-1+IFN-α联合用药比TT-1单独给药更能有效地抑制HepG-2/Huh7移植瘤的生长,但是在消除NK细胞之后该效应明显减弱,说明TT-1+IFN-α的抗肿瘤效应是通过NK细胞特异性介导的。TT-1不仅可以上调肿瘤细胞表面MICA的表达量,而且可以减少可溶性MICA的分泌;进一步研究表明,TT-1通过抑制去整合素金属蛋白酶10(ADAM 10)的表达来阻止MICA从肿瘤细胞表面脱落。细胞毒性实验表明,TT-1+IFN-α可以显著增强NK细胞对HepG-2/Huh7细胞的杀伤作用。免疫组化实验结果显示,TT-1+IFN-α联合用药可以明显增加肿瘤组织中肿瘤细胞表面MICA和NK细胞NKG2D的表达量。综上所述,TT-1+IFN-α联合用药可以通过增强MICA和NKG2D的相互作用达到显著的抗肿瘤效果。

关键词:TT-1;干扰素α;自然杀伤细胞;肝癌;免疫治疗

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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