聚乙二醇化干扰素α2A抑制人肝癌细胞增殖在体外和体内

StephenW

PLoS One. 2013; 8(12): e83195.
Published online 2013 December 12. doi:  10.1371/journal.pone.0083195
PMCID: PMC3861497
Pegylated Interferon-α2a Inhibits Proliferation of Human Liver Cancer Cells In Vitro and In Vivo
Hironori Kusano,1,* Jun Akiba,1 Sachiko Ogasawara,1 Sakiko Sanada,1 Makiko Yasumoto,1 Masamichi Nakayama,1 Keiko Ueda,1 Kosuke Ueda,1 Takashi Kurita,1 Keita Todoroki,1 Yumi Umeno,1 Osamu Nakashima,2 and Hirohisa Yano1
Diego Calvisi, Editor
1Department of Pathology, Kurume University School of Medicine, Kurume, Fukuoka, Japan
2Department of Clinical Laboratory Medicine, Kurume University Hospital, Kurume, Fukuoka, Japan
Institut für Pathologie, Greifswald, Germany, Germany
* E-mail: [email protected]
Competing Interests: The authors have declared that no competing interests exist.

Conceived and designed the experiments: HK JA SO ON HY. Performed the experiments: HK JA SO SS MY MN KU KU TK KT YU ON HY. Analyzed the data: HK JA SO SS MY MN KU KU TK KT YU ON HY. Contributed reagents/materials/analysis tools: HK JA SO SS MY MN KU KU TK KT YU ON HY. Wrote the manuscript: HK JA SO ON HY.

Abstract
Purpose

We investigated the effects of pegylated interferon-α2a (PEG-IFN-α2a) on the growth of human liver cancer cells.
Methods

The effect of PEG-IFN-α2a on the proliferation of 13 liver cancer cell lines was investigated in vitro. Cells were cultured with medium containing 0–4,194 ng/mL of PEG-IFN-α2a, and after 1, 2, 3, or 4 days of culture, morphologic observation and growth assay were performed. After hepatocellular carcinoma (HCC) cells (HAK-1B and KIM-1) were transplanted into nude mice, various doses of PEG-IFN-α2a were subcutaneously administered to the mice once a week for 2 weeks, and tumor volume, weight, and histology were examined.
Results

PEG-IFN-α2a inhibited the growth of 8 and 11 cell lines in a time- and dose-dependent manner, respectively, although the 50% growth inhibitory concentrations of 7 measurable cell lines on Day 4 were relatively high and ranged from 253 ng/mL to 4,431 ng/mL. Various levels of apoptosis induction were confirmed in 8 cell lines. PEG-IFN-α2a induced a dose-dependent decrease in tumor volume and weight, and a significant increase of apoptotic cells in the tumor. Subcutaneous administration of clinical dose for chronic hepatitis C (3 μg/kg, 0.06 μg/mouse) was effective and induced about 30-50% reduction in the tumor volume and weight as compared with the control.
Conclusions

Although in vitro anti-proliferative effects of PEG-IFN-α2a were relatively weak, PEG-IFN-α2a induced strong anti-tumor effects on HCC cells in vivo. The data suggest potential clinical application of PEG-IFN-α2a for the prevention and treatment of HCC.

Introduction

Interferons (IFNs) are types of cytokine that are produced by host cells, such as leukocytes, in response to inflammation. Since IFNs possess antiviral activity, antiproliferative activity and various immunoregulatory activities, IFN therapy is used to treat patients with chronic viral hepatitis or certain types of cancer including malignant melanoma, acquired immunodeficiency syndrome-related Kaposi’s sarcoma and some hematopoietic malignancies [1,2]. Lai et al also showed that recombinant IFNα is useful in prolonging survival among patients with inoperable hepatocellular carcinoma (HCC) [3]. In addition, some studies showed IFN therapy might prevent either occurrence or recurrence after initial curative therapy of HCC, such as liver resection and radiofrequency ablation, in patient with chronic viral hepatitis [4–7]. This cancer preventive effect of IFNs is regarded mainly as results of their antiviral effect and the consequent suppression of inflammation, and might be due to their direct antitumor effect against clinically undetectable HCC as well. The detailed mechanism of the antitumor effect of IFNs, however, remains obscure.

Pegylated interferon-α2a (PEG-IFN-α2a) and pegylated interferon-α2b (PEG-IFN-α2b), which are used to treat patients with chronic hepatitis C virus (HCV) or B virus (HBV) infection, are modified IFNs that have longer serum half-life in body than non-pegylated forms of IFNs, therefore they can be given to patients only once a week, whereas a standard IFN without pegylation used to be injected up to three to five times a week. This once-a-week injection of pegylated IFNs in combination with daily oral dosing of the nucleoside analogue ribavirin has substantially improved the rate of sustained virological response in patients with chronic HCV infection and got a position as the first line therapy [8,9]. We previously reported that PEG-IFN-α2b which contains 12 kDa polyethylene glycol (PEG) has stronger antitumor effects in vivo than non-pegylated IFNs and this result might be indicating that continuous IFNs exposure to cancer cells in body is more effective than continual injection [10]. On the basis of above-described background, we examined the growth inhibitory effects of PEG-IFN-α2a which contains two chains of 20 kDa PEG and has the longest serum half-life among clinically available IFNs on liver cancer cell lines in vitro and in vivo.

StephenW

公共科学图书馆之一。 2013 ， 8 （ 12 ） ： e83195 。
网上公布的2013年12月12。 DOI ： 10.1371/journal.pone.0083195
PMCID ： PMC3861497
聚乙二醇化干扰素α2A抑制人肝癌细胞增殖在体外和体内
草野弘典， 1 ， *君秋叶， 1小笠原祥子， 1真田咲子， 1安本真纪子， 1中山正道， 1惠子上田， 1介上田， 1栗田隆史， 1凯塔轰， 1美梅野， 1修中岛， 2和裕久Yano1
迭戈Calvisi ，编辑器
病理学教研室的，久留米大学医学院，久留米，福冈，日本
临床检验医学，久留米大学医院，久留米，福冈，日本教研室
研究所献给Pathologie ，德国Greifswald ，德国
*电子邮箱： [email protected]
竞争利益：作者声明：不存在利益竞争。

构思和设计的实验：港币JA等HY 。进行的实验：港币JA SO SS我的明尼苏达KU KU TK KT羽对HY 。分析数据：香港JA SO SS我的明尼苏达KU KU TK KT羽对HY 。贡献试剂/材料/分析工具：港币JA SO SS我的明尼苏达KU KU TK KT羽对HY 。写的稿子：港币JA等HY 。

摘要
目的

我们研究了聚乙二醇化干扰素- α2A （ PEG-IFN- α2A ）对人肝癌细胞生长的影响。
方法

PEG-IFN- α2A对13肝癌细胞系增殖的影响进行了研究体外。细胞培养用含PEG-IFN- α2A的0-4,194纳克/毫升培养基中， 1，2， 3，或4天培养，形态观察及生长测定被执行之后。肝细胞癌后（HCC）的细胞（ HAK -1B和KIM- 1）被移植到裸鼠，不同剂量的PEG-IFN- α2A的皮下给药至小鼠，每周一次， 2周后，肿瘤体积，重量和组织学检查。
结果

PEG-IFN- α2A抑制的8和11的细胞系中以时间和剂量依赖的方式，分别生长，虽然50％的生长抑制浓度的第4天7可测量的细胞系是相对较高的，并从253纳克/ mL至4,431毫微克/毫升。诱导细胞凋亡的各级8细胞系中证实。 PEG-IFN- α2A诱导的剂量依赖性减少肿瘤的体积和重量，以及凋亡细胞在肿瘤显著增加。临床剂量用于治疗慢性丙型肝炎（ 3微克/公斤， 0.06微克/小鼠）皮下给药是有效的和诱导的肿瘤体积和体重，与对照相比，减少约30-50 ％。
结论

虽然在体外抗增殖PEG-IFN- α2A的影响是相对较弱， PEG-IFN- α2A诱导对肝癌细胞在体内强大的抗肿瘤作用。这些数据表明肝癌的预防和治疗潜在的临床应用PEG-IFN - α2A的。

介绍

干扰素（干扰素）是类型的细胞因子是由宿主细胞，如白细胞，在对炎症的反应制备。自干扰素具有抗病毒活性，抗增殖活性和各种免疫调节活性，干扰素治疗是用于治疗慢性病毒性肝炎或某些类型的癌症，包括恶性黑色素瘤，获得性免疫缺陷综合征有关的卡波济氏肉瘤以及一些造血系统恶性肿瘤[1,2]。 Lai等人还发现，重组干扰素α是在延长的患者不能手术的肝细胞癌（HCC ）的生存有用的[ 3 ] 。此外，一些研究表明，干扰素治疗可能会阻止任何发生或复发肝癌的疗效初步治疗后，如肝切除术和射频消融治疗，在病人与慢性病毒性肝炎[ 4-7 ] 。 IFN的这种癌症的预防作用主要是作为其抗病毒效果和炎症的结果抑制效果，并且可能是由于对临床检测不到的肝癌其直接的抗肿瘤效果为好。的干扰素的抗肿瘤作用的详细机制，但是，仍然不清楚。

聚乙二醇化干扰素- α2A （ PEG-IFN- α2A ）和聚乙二醇化的干扰素- α2b的（ PEG-IFN- α2b的），它是用于治疗慢性丙型肝炎病毒（HCV）或乙型肝炎病毒（HBV）感染，被修饰的干扰素是有较长的血清半衰期在身体比IFN的非聚乙二醇化形式，因此它们可以被给予患者每周一次，而标准的IFN未经PEG化用于将注入到三到五倍一个星期。这一次的周注射与核苷类似物利巴韦林的每日口服剂量组合干扰素聚乙二醇化已大大改善患者的慢性丙型肝炎病毒感染持续病毒学应答率，并得到了一个位置作为第一线治疗[ 8,9] 。我们以前报道， PEG-IFN- α2b的其中含有12 kDa的聚乙二醇（PEG ）具有在体内比非PEG化的干扰素更强的抗肿瘤作用和该结果可能表明干扰素连续暴露于癌细胞在身体比连续注射更有效[10] 。在上述背景的基础上，我们研究了PEG-IFN- α2A其中包含的20kDa PEG的两条链，并具有最长的血清中对在体外和肝癌细胞株临床上可用干扰素半衰期的增殖抑制效果体内。