Synergistic effect of cytokine-induced killer cell with valproate inhibits growth of hepatocellular carcinoma cell in a mouse model
Synergistic effect of cytokine-induced killer cell with valproate inhibits growth of hepatocellular carcinoma cell in a mouse model
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DOI：
10.1080/15384047.2016.1276132
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发表时间：
2017-01-01
影响因子：
3.6
通讯作者：
Yoon, Jung-Hwan
中科院分区：
文献类型：
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作者：
Lee, Dong Hyeon;Nam, Joon Yeul;Yoon, Jung-Hwan
目的：由于缺乏治疗方案，晚期肝细胞癌（HCC）的长期预后仍然很差。细胞因子诱导的杀伤细胞（CIK）是体外扩增的T淋巴细胞，表达NK和T细胞标记。仅CIK细胞疗法不足以治疗晚期HCC。因此，本研究旨在确定CIK细胞联合丙戊酸（VPA）治疗是否能提供协同作用，抑制小鼠肝癌模型的肿瘤生长。方法：采用流式细胞术（FACS）评价自然杀伤组2D （NKG2D）配体（维甲酸早期诱导1 [RAE-1]，小鼠；主要组织相容性复合体I类多肽相关序列A [MIC-A]，人）的上调。计算不减小肿瘤体积的VPA浓度，以避免VPA在C3H小鼠HCC模型中的细胞毒性。CIK细胞是用干扰素γ、CD3单克隆抗体和白细胞介素2从小鼠脾细胞中生成的。在小鼠模型中评价CIK细胞与VPA联合的潜在协同作用，并进行组织病理学研究。结果：与VPA孵育40h后，4株HCC细胞株中RAE-1和MIC-A的表达均较对照组增加（MH-134增加2.3倍，Huh-7增加2.4倍，SNU-761增加3.7倍，SNU-475增加6.5倍）。与对照组相比，VPA的最大体内剂量为10mg/kg/d，未显示出显著的细胞毒性。C3H小鼠脾细胞可生成CIK细胞。CIK细胞加VPA治疗7d后，对小鼠肝癌模型的相对肿瘤体积有增效作用。对照组小鼠的相对肿瘤体积增加到11.25，而联合治疗组仅增加到5.20 （P = 0.047）。结论：vpa诱导的NKG2D配体表达的增加显著增强了CIK细胞治疗小鼠肝癌模型的效果。
Objective: Long-term prognosis of hepatocellular carcinoma (HCC) remains poor owing to the lack of treatment options for advanced HCC. Cytokine-induced killer (CIK) cells are ex vivo expanded T lymphocytes expressing both NK- and T-cell markers. CIK cell therapy alone is insufficient for treating advanced HCC. Thus, this study aimed to determine whether treatment with CIK cells combined with valproic acid (VPA) could provide a synergistic effect to inhibit tumor growth in a mouse model of HCC. Methods: Upregulation of natural killer group 2D (NKG2D) ligands (retinoic acid early inducible 1 [RAE-1], mouse; major histocompatibility complex class I polypeptide-related sequence A [MIC-A], human) were evaluated by FACS. VPA concentrations that did not reduce tumor volume were calculated to avoid VPA cytotoxicity in a C3H mouse model of HCC. CIK cells were generated from mouse splenocytes using interferon gamma, a CD3 monoclonal antibody, and interleukin 2. The potential synergistic effect of CIK cells combined with VPA was evaluated in the mouse model and tissue pathology was investigated. Results: After 40h of incubation with VPA, RAE-1 and MIC-A expression were increased in 4 HCC cell lines compared with that in control (2.3-fold in MH-134, 2.4-fold in Huh-7, 3.7-fold in SNU-761, and 6.5-fold in SNU-475). The maximal in vivo VPA dosage that showed no significant cytotoxicity compared with control was 10mg/kg/day. CIK cells were well generated from C3H mouse splenocytes. After 7d of treatment with CIK cells plus VPA, a synergistic effect was observed on relative tumor volume in the mouse model of HCC. While the relative tumor volume in untreated control mice increased to 11.25, that in the combination treatment group increased to only 5.20 (P = 0.047). Conclusions: The VPA-induced increase in NKG2D ligands expression significantly enhanced the effects of CIK cell therapy in a mouse model of HCC.