香菇多糖联合AdIRF3通过刺激IFN-β表达抑制乳腺癌生长
|
摘要
目的利用荷瘤小鼠模型探讨香菇多糖联合干扰素调节因子3(IRF3)表达腺病毒载体治疗肿瘤的价值及潜在的机制。方法利用荷瘤Balb/c小鼠,分别接种磷酸盐缓冲液(PBS)、空白对照腺病毒载体(Ad GFP)、香菇多糖、表达活化型IRF3的腺病毒载体(Ad IRF3)及香菇多糖联合Ad IRF3,观察荷瘤小鼠的肿瘤生长曲线;利用Real-time PCR检测肿瘤组织的β干扰素(IFN-β)水平,酶联免疫斑点检测法(ELISPOT)检测脾脏淋巴细胞功能,流式细胞术(FACS)检测肿瘤局部淋巴细胞比例;记录荷瘤Balb/c裸鼠的肿瘤生长曲线,以及α干扰素受体1(IFNAR1)抗体封闭Balb/c的IFNAR后肿瘤生长曲线。结果与对照组相比,香菇多糖联合Ad IRF3可抑制Balb/c荷瘤小鼠肿瘤生长,且肿瘤局部组织中IFN-β显著高于对照组;肿瘤组织浸润CD8+T细胞比例显著高于对照组,而调节性T细胞(Treg)细胞比例在各组间并无显著性差异;ELISPOT显示联合处理刺激淋巴细胞分泌γ干扰素(IFN-γ)的能力显著高于对照组;荷瘤Balb/c裸鼠在接受不同组处理后,各组间的肿瘤生长曲线并无显著差异;利用IFNAR1封闭抗体阻断干扰素作用后,荷瘤Balb/c小鼠再次接受不同组处理,结果显示封闭抗体处理后可抑制香菇多糖联合Ad IRF3的肿瘤抑制作用。结论香菇多糖联合Ad IRF3通过活化IRF3-IFN通路发挥协同抑制乳腺癌肿瘤生长的作用。
Objective To investigate the effect of lentinan(LNT) combined with IRF3 expressing adenovirus vector on the anti-tumor therapy, and its underlying mechanism. Methods Balb/c bearing TUBO mice were treated with PBS, AdGFP, AdIRF3,LNT, and LNT combined with AdIRF3 to observe the tumor growth curve of mice. qPCR was used to detect the IFN-β level of tumor tissues. ELISPOT was to check the IFN-γ secretion of spleen cells. FACS was used to make sure the percentage of tumor infiltrated T cells. Balb/c-nu mice were inoculated with TUBO cells, and tumor growth was monitored. Anti-IFNAR1 antibody was injected to Babl/c bearing mice, and tumor growth was measured. Results Compared with control groups, LNT combined with AdIRF3 can inhibit the tumor growth and increase the IFN-γ secretion of TUBO-bearing mice. There was no significant difference of the percentage of tumor infiltrated Treg among groups, but the tumor infiltrated CD8+T was significantly higher in LNT combined with Ad IRF3 treated mice group. There was no significant difference of tumor growth between Balb-nu mice inoculated with TUBO cells treated by LNT/AdIRF3 and PBS/AdGFP group. The tumor growth was not inhibited in Babl/c bearing mice injected with anti-IFNAR1 antibody and treated by LNT/AdIRF3. Conclusion LNT combined with AdIRF3 can inhibit the tumor growth by activating the IRF3-IFN signaling pathway.
Objective To investigate the effect of lentinan(LNT) combined with IRF3 expressing adenovirus vector on the anti-tumor therapy, and its underlying mechanism. Methods Balb/c bearing TUBO mice were treated with PBS, AdGFP, AdIRF3,LNT, and LNT combined with AdIRF3 to observe the tumor growth curve of mice. qPCR was used to detect the IFN-β level of tumor tissues. ELISPOT was to check the IFN-γ secretion of spleen cells. FACS was used to make sure the percentage of tumor infiltrated T cells. Balb/c-nu mice were inoculated with TUBO cells, and tumor growth was monitored. Anti-IFNAR1 antibody was injected to Babl/c bearing mice, and tumor growth was measured. Results Compared with control groups, LNT combined with AdIRF3 can inhibit the tumor growth and increase the IFN-γ secretion of TUBO-bearing mice. There was no significant difference of the percentage of tumor infiltrated Treg among groups, but the tumor infiltrated CD8+T was significantly higher in LNT combined with Ad IRF3 treated mice group. There was no significant difference of tumor growth between Balb-nu mice inoculated with TUBO cells treated by LNT/AdIRF3 and PBS/AdGFP group. The tumor growth was not inhibited in Babl/c bearing mice injected with anti-IFNAR1 antibody and treated by LNT/AdIRF3. Conclusion LNT combined with AdIRF3 can inhibit the tumor growth by activating the IRF3-IFN signaling pathway.
引文
[1]Hu Z I,McArthur H L.Immunotherapy in breast cancer:The new frontier[J].Curr Breast Cancer Rep,2018,10(2):35-40.
[2]Parvizpour S,Razmara J,Omidi Y.Breast cancer vaccination comes to age:Impacts of bioinformatics[J].Bioimpacts,2018,8(3):223-235.
[3]Heylbroeck C,Balachandran S,Servant M J,et al.The IRF-3 transcription factor mediates Sendai virus-induced apoptosis[J].J Virol,2000,74(8):3781-3792.
[4]Pencheva N,de Gooijer M C,Vis D J,et al.Identification of a druggable pathway controlling glioblastoma invasiveness[J].Cell Rep,2017,20(1):48-60.
[5]Ysebrant D L L,Martinet V,Goriely S.Interferon regulatory factor 3 in adaptive immune responses[J].Cell Mol Life Sci,2014,71(20):3873-3883.
[6]Hochmann S,Mittermeir M,Santic R,et al.Evaluation of modified Interferon alpha mRNA constructs for the treatment of non-melanoma skin cancer[J].Sci Rep,2018,doi:10.1038/s41598-018-31061-w.
[7]Yi W,Zhang P,Hou J,et al.Enhanced response of tamoxifen toward the cancer cells using a combination of chemotherapy and photothermal ablation induced by lentinan-functionalized multi-walled carbon nanotubes[J].Int J Biol Macromol,2018,120(Pt B):1525-1532.
[8]Wang H,Cai Y,Zheng Y,et al.Efficacy of biological response modifier lentinan with chemotherapy for advanced cancer:A meta-analysis[J].Cancer Med,2017,6(10):2222-2233.
[9]李石军,王凯平,汪柳,等.香菇多糖LNT2的提取分离纯化、结构及体外抗肿瘤活性研究[J].中草药,2014,45(9):1232-1237.
[10]Deng S,Zhang G,Kuai J,et al.Lentinan inhibits tumor angiogenesis via interferon gamma and in a T cell independent manner[J].J Exp Clin Cancer Res,2018,doi:10.1186/s13046-018-0932-y.
[11]Dubrot J,Palazon A,Alfaro C,et al.Intratumoral injection of interferon-alpha and systemic delivery of agonist anti-CD137 monoclonal antibodies synergize for immunotherapy[J].Int J Cancer,2011,128(1):105-118.
[12]Liang Y,Tang H,Guo J,et al.Targeting IFNalpha to tumor by anti-PD-L1 creates feedforward antitumor responses to overcome checkpoint blockade resistance[J].Nat Commun,2018,doi:10.1038/s41467-018-06890-y.
[13]Heo J,Reid T,Ruo L,et al.Randomized dose-finding clinical trial of oncolytic immunotherapeutic vaccinia JX-594 in liver cancer[J].Nat Med,2013,19(3):329-336.
[14]Park B H,Hwang T,Liu T C,et al.Use of a targeted oncolytic poxvirus,JX-594,in patients with refractory primary or metastatic liver cancer:A phase I trial[J].Lancet Oncol,2008,9(6):533-542.
[15]Yanai H,Chiba S,Hangai S,et al.Revisiting the role of IRF3 in inflammation and immunity by conditional and specifically targeted gene ablation in mice[J].Proc Natl Acad Sci USA,2018,115(20):5253-5258.
[16]Chen Q,Peng H,Dong L,et al.Activation of the NRF2-ARE signalling pathway by the Lentinula edodes polysaccharose LNT alleviates ROS-mediated cisplatin nephrotoxicity[J].Int Immunopharmacol,2016,36:1-8.
[17]Katlinskaya Y V,Katlinski K V,Yu Q,et al.Suppression of type I interferon signaling overcomes oncogene-induced senescence and mediates melanoma development and progression[J].Cell Rep,2016,15(1):171-180.
[18]Katlinski K V,Gui J,Katlinskaya Y V,et al.Inactivation of interferon receptor promotes the establishment of immune privileged tumor microenvironment[J].Cancer Cell,2017,31(2):194-207.
[2]Parvizpour S,Razmara J,Omidi Y.Breast cancer vaccination comes to age:Impacts of bioinformatics[J].Bioimpacts,2018,8(3):223-235.
[3]Heylbroeck C,Balachandran S,Servant M J,et al.The IRF-3 transcription factor mediates Sendai virus-induced apoptosis[J].J Virol,2000,74(8):3781-3792.
[4]Pencheva N,de Gooijer M C,Vis D J,et al.Identification of a druggable pathway controlling glioblastoma invasiveness[J].Cell Rep,2017,20(1):48-60.
[5]Ysebrant D L L,Martinet V,Goriely S.Interferon regulatory factor 3 in adaptive immune responses[J].Cell Mol Life Sci,2014,71(20):3873-3883.
[6]Hochmann S,Mittermeir M,Santic R,et al.Evaluation of modified Interferon alpha mRNA constructs for the treatment of non-melanoma skin cancer[J].Sci Rep,2018,doi:10.1038/s41598-018-31061-w.
[7]Yi W,Zhang P,Hou J,et al.Enhanced response of tamoxifen toward the cancer cells using a combination of chemotherapy and photothermal ablation induced by lentinan-functionalized multi-walled carbon nanotubes[J].Int J Biol Macromol,2018,120(Pt B):1525-1532.
[8]Wang H,Cai Y,Zheng Y,et al.Efficacy of biological response modifier lentinan with chemotherapy for advanced cancer:A meta-analysis[J].Cancer Med,2017,6(10):2222-2233.
[9]李石军,王凯平,汪柳,等.香菇多糖LNT2的提取分离纯化、结构及体外抗肿瘤活性研究[J].中草药,2014,45(9):1232-1237.
[10]Deng S,Zhang G,Kuai J,et al.Lentinan inhibits tumor angiogenesis via interferon gamma and in a T cell independent manner[J].J Exp Clin Cancer Res,2018,doi:10.1186/s13046-018-0932-y.
[11]Dubrot J,Palazon A,Alfaro C,et al.Intratumoral injection of interferon-alpha and systemic delivery of agonist anti-CD137 monoclonal antibodies synergize for immunotherapy[J].Int J Cancer,2011,128(1):105-118.
[12]Liang Y,Tang H,Guo J,et al.Targeting IFNalpha to tumor by anti-PD-L1 creates feedforward antitumor responses to overcome checkpoint blockade resistance[J].Nat Commun,2018,doi:10.1038/s41467-018-06890-y.
[13]Heo J,Reid T,Ruo L,et al.Randomized dose-finding clinical trial of oncolytic immunotherapeutic vaccinia JX-594 in liver cancer[J].Nat Med,2013,19(3):329-336.
[14]Park B H,Hwang T,Liu T C,et al.Use of a targeted oncolytic poxvirus,JX-594,in patients with refractory primary or metastatic liver cancer:A phase I trial[J].Lancet Oncol,2008,9(6):533-542.
[15]Yanai H,Chiba S,Hangai S,et al.Revisiting the role of IRF3 in inflammation and immunity by conditional and specifically targeted gene ablation in mice[J].Proc Natl Acad Sci USA,2018,115(20):5253-5258.
[16]Chen Q,Peng H,Dong L,et al.Activation of the NRF2-ARE signalling pathway by the Lentinula edodes polysaccharose LNT alleviates ROS-mediated cisplatin nephrotoxicity[J].Int Immunopharmacol,2016,36:1-8.
[17]Katlinskaya Y V,Katlinski K V,Yu Q,et al.Suppression of type I interferon signaling overcomes oncogene-induced senescence and mediates melanoma development and progression[J].Cell Rep,2016,15(1):171-180.
[18]Katlinski K V,Gui J,Katlinskaya Y V,et al.Inactivation of interferon receptor promotes the establishment of immune privileged tumor microenvironment[J].Cancer Cell,2017,31(2):194-207.