Curcumin plays a synergistic role in combination with HSV-TK/GCV in inhibiting growth of murine B16 melanoma cells and melanoma xenografts
Curcumin plays a synergistic role in combination with HSV-TK/GCV in inhibiting growth of murine B16 melanoma cells and melanoma xenografts
复制标题
姜黄素与HSV-TK/GCV联合抑制小鼠B16黑色素瘤细胞和黑色素瘤异种移植物的生长发挥协同作用
DOI：
10.7717/peerj.7760
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发表时间：
2019-09-20
期刊：
影响因子：
2.7
通讯作者：
Tan, Yuhui
中科院分区：
文献类型：
--
作者：
Li, Hong;Du, Haiyan;Tan, Yuhui
黑色素瘤是全球关注的问题，是皮肤癌的主要死亡原因。单纯疱疹病毒胸苷激酶基因联合更昔洛韦（HSV-TK/GCV）是一种很有前途的黑色素瘤基因治疗方法。尽管其效率低，但众所周知的是其主要由间隙连接介导的旁观者效应。在这项研究中，我们发现姜黄素以时间和剂量依赖性方式降低B16黑色素瘤细胞的活力。进一步的研究表明，姜黄素可增强HSV-TK/GCV的差距连接细胞间通讯（GJIC）功能，上调间隙连接蛋白32和43等间隙连接蛋白的表达，可能在增强HSV-TK/GCV的旁观者效应中发挥作用。通过将稳定表达TK基因的B16 TK细胞与野生型B16（B16 WT）细胞共培养，我们发现姜黄素和GCV联合处理可协同抑制B16细胞增殖，但这种作用可被差距连接抑制剂阿加消除。姜黄素可显著增加B16 WT细胞的凋亡率，提示姜黄素可增强HSV-TK/GCV的旁观者效应。在体内研究中，我们通过以3：7的比例注射B16 TK和B16 WT细胞的混合物，在14天内建立了异种移植的黑素瘤模型。结果表明，姜黄素和GCV联合给药显著抑制了异种移植物的生长，如较小的尺寸和较轻的重量所示。进一步证实了组合效应为协同效应。结论：姜黄素可增强HSV-TK/GCV对黑色素瘤的杀伤作用和旁观者效应，其机制可能与改善细胞间隙连接有关。我们的数据表明，HSV-TK/GCV与姜黄素的组合可能是一种潜在的癌症治疗的化学增敏策略。
Melanoma is a global concern and accounts for the major mortality of skin cancers. Herpes simplex virus thymidine kinase gene with ganciclovir (HSV-TK/GCV) is a promising gene therapy for melanoma. Despite its low efficiency, it is well known for its bystander effect which is mainly mediated by gap junction. In this study, we found that curcumin reduced B16 melanoma cell viability in both time- and dose-dependent manner. Further study showed that curcumin improved the gap junction intercellular communication (GJIC) function, and upregulated the proteins essential to gap junction, such as connexin 32 and connexin 43, indicating the potential role in enhancing the bystander effect of HSV-TK/GCV. By co-culturing the B16TK cells, which stably expressed TK gene, with wildtype B16 (B16WT) cells, we found that co-treatment of curcumin and GCV synergistically inhibited B16 cell proliferation, but the effect could be eliminated by the gap junction inhibitor AGA. Moreover, curcumin markedly increased apoptosis rate of B16WT cells, suggesting its effect in enhancing the bystander effect of HSV-TK/GCV. In the in-vivo study, we established the xenografted melanoma model in 14 days by injecting mixture of B16TK and B16WT cell in a ratio of 3:7. The result demonstrated that, co-administration of curcumin and GCV significantly inhibited the xenograft growth, as indicated by the smaller size and less weight. The combinational effect was further confirmed as a synergistic effect. In conclusion, the results demonstrated that curcumin could enhance the killing effect and the bystander effect of HSV-TK/GCV in treating melanoma, which might be mediated by improved gap junction. Our data suggested that combination of HSV-TK/GCV with curcumin could be a potential chemosensitization strategy for cancer treatment.