Oncolytic influenza virus enhances killing effect of gemcitabine against pancreatic cancer cells—MENG Zihe1,2, XU Yongru2, DENG Zhaoda2, ZHANG Yuxin1,2, YANG Penghui1,2, HU Ruiping1 1College of Basic Medical Sciences, Inner Mongolia Medical University, Hohhot, Inner Mongolia; 2Institute of Hepato-Pancreato-Biliary Surgery, First Medical Center, Chinese PLA General Hospital, Beijing, China

doi:10.16016/j.2097-0927.202407052

您当前的位置：

首页 >

文章列表页 >

Oncolytic influenza virus enhances killing effect of gemcitabine against pancreatic cancer cells—MENG Zihe^1,2, XU Yongru², DENG Zhaoda², ZHANG Yuxin^1,2, YANG Penghui^1,2, HU Ruiping¹ ¹College of Basic Medical Sciences, Inner Mongolia Medical University, Hohhot, Inner Mongolia;²Institute of Hepato-Pancreato-Biliary Surgery, First Medical Center, Chinese PLA General Hospital, Beijing, China

Basic Medicine | 更新时间：2025-01-23

- Oncolytic influenza virus enhances killing effect of gemcitabine against pancreatic cancer cells—MENG Zihe^1,2, XU Yongru², DENG Zhaoda², ZHANG Yuxin^1,2, YANG Penghui^1,2, HU Ruiping¹ ¹College of Basic Medical Sciences, Inner Mongolia Medical University, Hohhot, Inner Mongolia;²Institute of Hepato-Pancreato-Biliary Surgery, First Medical Center, Chinese PLA General Hospital, Beijing, China
- Journal of Army Medical University Vol. 47, Issue 2, Pages: 141-150(2025)
- 作者机构：
  
  1.内蒙古医科大学基础医学院，内蒙古呼和浩特
  2.中国人民解放军总医院第一医学中心肝胆胰外科医学部研究所，北京
- 作者简介：
- 基金信息：
- DOI：10.16016/j.2097-0927.202407052
  CLC：
- Published：30 January 2025，
  
  Received：12 July 2024，
  
  Revised：04 December 2024，
- 稿件说明：
移动端阅览
Oncolytic influenza virus enhances killing effect of gemcitabine against pancreatic cancer cells—MENG Zihe^1,2, XU Yongru², DENG Zhaoda², ZHANG Yuxin^1,2, YANG Penghui^1,2, HU Ruiping¹ ¹College of Basic Medical Sciences, Inner Mongolia Medical University, Hohhot, Inner Mongolia;²Institute of Hepato-Pancreato-Biliary Surgery, First Medical Center, Chinese PLA General Hospital, Beijing, China. [J]. Journal of army medical university, 2025, 47(2): 141-150.
DOI：

Oncolytic influenza virus enhances killing effect of gemcitabine against pancreatic cancer cells—MENG Zihe^1,2, XU Yongru², DENG Zhaoda², ZHANG Yuxin^1,2, YANG Penghui^1,2, HU Ruiping¹ ¹College of Basic Medical Sciences, Inner Mongolia Medical University, Hohhot, Inner Mongolia;²Institute of Hepato-Pancreato-Biliary Surgery, First Medical Center, Chinese PLA General Hospital, Beijing, China. [J]. Journal of army medical university, 2025, 47(2): 141-150. DOI： 10.16016/j.2097-0927.202407052.

摘要

目的

探究基于反向遗传学（reversed genetics，RG）技术构建的重组溶瘤流感病毒OvFlu-GM-CSF联合化疗药吉西他滨（gemcitabine，GEM）杀伤胰腺癌细胞的效果。

方法

前期利用RG技术成功拯救重组溶瘤病毒OvFlu-GM-CSF，通过鸡红细胞凝集实验、透射电镜、病毒复制等对OvFlu-GM-CSF进行全面鉴定，CCK-8法检测OvFlu-GM-CSF在不同时间（24、48、72 h）、不同感染复数（MOI=0、0.1、1.0、3.0）条件下，对胰腺癌细胞系（Panc02、PANC-1、SW1990、BxPC-3）及正常胰腺导管上皮细胞（HPDE6-C7）存活率的影响。利用胰腺癌小鼠皮下荷瘤模型，按照完全随机法将6周龄雌性C57BL/6小鼠分为4组（

=9）：PBS组、重组溶瘤病毒组（OvFlu-GM-CSF组）、吉西他滨组（GEM组）以及重组溶瘤病毒与吉西他滨联合治疗组（OvFlu-GM-CSF+GEM组），并分别给予PBS、OvFlu-GM-CSF、GEM、OvFlu-GM-CSF+GEM治疗，监测小鼠肿瘤体积及存活率等指标的变化。采用多重免疫荧光染色技术分析肿瘤组织中T细胞浸润及增殖情况。通过HE染色检测小鼠主要器官（心脏、肝脏、肺脏、肾脏、脑组织）的病理变化，小鼠血清肝功能实验检测谷丙转氨酶（ALT）和谷草转氨酶（AST）酶活性水平，以评估重组溶瘤病毒的安全性。

结果

重组溶瘤流感病毒OvFlu-GM-CSF的血凝效价为2

，其病毒形态符合流感病毒典型特征，且病毒在胰腺癌细胞内可选择性复制。在细胞水平，OvFlu-GM-CSF在感染时间为48 h、MOI=3.0的条件下，与感染时间为48 h、MOI=0比较，对Panc02、PANC-1、SW1990、BxPC-3具有显著的选择性杀伤作用（

0.01），且细胞存活率随着MOI值的增加以及感染时间的延长而逐渐降低（

0.01），但对HPDE6-C7无明显影响。在胰腺癌荷瘤小鼠体内，与PBS组的肿瘤体积比较，OvFlu-GM-CSF组、OvFlu-GM-CSF+GEM组显著减小（

0.01）。多重免疫荧光检测结果显示在OvFlu-GM-CSF+GEM组小鼠的肿瘤组织中T淋巴细胞浸润显著增加；在小鼠心脏、肝脏、肺脏、肾脏、脑组织中未观察到明显病理变化；与PBS组比较，OvFlu-GM-CSF组、GEM组、OvFlu-GM-CSF+GEM组小鼠血清中ALT及AST酶活性水平无明显差异。

结论

通过反向遗传学技术构建的重组溶瘤流感病毒OvFlu-GM-CSF与化疗药GEM联合应用，提升了对胰腺癌细胞的杀伤效能，并有效激活了机体的抗肿瘤免疫应答。

Abstract

Objective‍　‍To investigate the killing effect of the recombinant oncolytic influenza virus OvFlu-GM-CSF， constructed using reverse genetics （RG） technology， in combination with chemotherapy drug， gemcitabine （GEM）， against pancreatic cancer cells. Methods‍　‍The recombinant oncolytic virus OvFlu-GM-CSF was successfully rescued using RG technology in our previous study. The virus was then comprehensively characterized through chicken red blood cell hemagglutination assay， transmission electron microscopy， and viral replication assay. CCK-8 assay was utilized to determine the impact of OvFlu-GM-CSF viruses ［multiplicities of infection （MOI）： 0， 0.1， 1.0， 3.0］ on the survival rate of pancreatic cancer cell lines （Panc02， PANC-1， SW1990， BxPC-3） and normal pancreatic ductal epithelial cells （HPDE6-C7） after treatment for 24， 48 or 72 h. Using a subcutaneous tumor-bearing mouse model of pancreatic cancer， 36 female C57BL/6 mice （6 weeks old） were randomly divided into PBS group， recombinant oncolytic virus group， GEM group， and the combined treatment group， with 9 mice in each group. PBS （100 μL/animal） or OvFlu-GM-CSF virus （1×10

PFU/100 μL） was given to the mice of the corresponding groups through intratumoral injection， while GEM （100 mg/kg） was injected intraperitoneally， once per 3 days， for totally 9 times. Changes in tumor volume and survival rate were monitored. Multi-immunofluorescence staining was employed to analyze T cell infiltration and proliferation in the tumor tissues. HE staining was performed to observe the pathological changes in major organs （heart， liver， lungs， kidneys and brain）， and the serum levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were measured to evaluate the safety of the recombinant oncolytic virus. Results‍　‍The recombinant oncolytic influenza virus OvFlu-GM-CSF has a hemagglutination titer of 2

， typical morphological features of influenza virus， and can selectively replicate within pancreatic cancer cells. At the cellular lev

el， the viruses demonstrated a significant selective cytotoxic effect on Panc02， PANC-1， SW1990， and BxPC-3 cells under the conditions of 48 h post-infection and MOI=3.0， when compared to 48 h post-infection and MOI=0 （

0.01）. The cell survival rate was gradually decreased with the increase in MOI value and the extension of infection time （

0.01）， but the viruses showed no significant effect on normal pancreatic ductal epithelial cells （HPDE6-C7）. In the pancreatic cancer tumor-bearing mouse model， the combined treatment of the viruses+GEM significantly reduced the tumor volume than simple virus treatment and simple GEM treatment （

0.01）， and enhanced the infiltration of T cells in the tumor tissues. No obvious pathological changes were observed in the above-mentioned major organs. Additionally， there were no significant differences in the serum levels of ALT and AST in the OvFlu-GM-CSF group， GEM group， and OvFlu-GM-CSF+GEM group compared to the PBS group. Conclusion‍　‍RS technology-constructed recombinant oncolytic influenza virus OvFlu-GM-CSF， when combined with the chemotherapeutic agent GEM， enhances the cytotoxic efficacy against pancreatic cancer cells and effectively activates the host’s anti-tumor immune response.

关键词

溶瘤流感病毒吉西他滨胰腺癌联合治疗

Keywords

oncolytic influenza virusgemcitabinepancreatic cancercombination therapy

references

MIZRAHI J， SURANA R， VALLE J， et al. Pancreatic cancer［J］. Lancet， 2020， 395： 2008-2020. DOI： 10.1016/S0140-6736（20）30974-0http://dx.doi.org/10.1016/S0140-6736（20）30974-0.

RAHIB L， SMITH B D， AIZENBERG R， et al. Projecting cancer incidence and deaths to 2030： the unexpected burden of thyroid， liver， and pancreas cancers in the United States［J］. Cancer Res， 2014， 74（11）： 2913-2921. DOI： 10.1158/0008-5472.CAN-14-0155http://dx.doi.org/10.1158/0008-5472.CAN-14-0155.

SIEGEL R， GIAQUINTO A N， JEMAL A. Cancer statistics， 2024［J］. CA Cancer J Clin， 2024， 74： 12-49. DOI： 10.3322/caac.21820http://dx.doi.org/10.3322/caac.21820.

KLEEFF J， KORC M， APTE M， et al. Pancreatic cancer［J］. Nat Rev Dis Primers， 2016， 2： 16022. DOI： 10.1038/nrdp.2016.22http://dx.doi.org/10.1038/nrdp.2016.22.

WOLFGANG C L， HERMAN J M， LAHERU D A， et al. Recent progress in pancreatic cancer［J］. CA Cancer J Clin， 2013， 63（5）： 318-348. DOI： 10.3322/caac.21190http://dx.doi.org/10.3322/caac.21190.

NEOPTOLEMOS J P， KLEEFF J， MICHL P， et al. Therapeutic developments in pancreatic cancer： current and future perspectives［J］. Nat Rev Gastroenterol Hepatol， 2018， 15（6）： 333-348. DOI： 10.1038/s41575-018-0005-xhttp://dx.doi.org/10.1038/s41575-018-0005-x.

STROBEL O， NEOPTOLEMOS J， JÄGER D， et al. Optimizing the outcomes of pancreatic cancer surgery［J］. Nat Rev Clin Oncol， 2019， 16（1）： 11-26. DOI： 10.1038/s41571-018-0112-1http://dx.doi.org/10.1038/s41571-018-0112-1.

HUANG L， JANSEN L， BALAVARCA Y， et al. Resection of pancreatic cancer in Europe and USA： an international large-scale study highlighting large variations［J］. Gut， 2019， 68（1）： 130-139. DOI： 10.1136/gutjnl-2017-314828http://dx.doi.org/10.1136/gutjnl-2017-314828.

HAMID O， HOFFNER B， GASAL E， et al. Oncolytic immunotherapy： unlocking the potential of viruses to help target cancer［J］. Cancer Immunol Immunother， 2017， 66（10）： 1249-1264. DOI： 10.1007/s00262-017-2025-8http://dx.doi.org/10.1007/s00262-017-2025-8.

JIN K T， DU W L， LIU Y Y， et al. Oncolytic virotherapy in solid tumors： the challenges and achievements［J］. Cancers， 2021， 13（4）： 588. DOI： 10.3390/cancers 13040588http://dx.doi.org/10.3390/cancers13040588.

DOUGAN M， DRANOFF G， DOUGAN S K. GM-CSF， IL-3， and IL-5 family of cytokines： regulators of inflammation［J］. Immunity， 2019， 50（4）： 796-811. DOI： 10.1016/j.immuni.2019.03.022http://dx.doi.org/10.1016/j.immuni.2019.03.022.

FERRUCCI P F， PALA L， CONFORTI F， et al. Talimogene laherparepvec （T-VEC）： an intralesional cancer immunotherapy for advanced melanoma［J］. Cancers， 2021， 13（6）： 1383. DOI： 10.3390/cancers 13061383http://dx.doi.org/10.3390/cancers13061383.

PACKIAM V T， LAMM D L， BAROCAS D A， et al. An open label， single-arm， phase II multicenter study of the safety and efficacy of CG0070 oncolytic vector regimen in patients with BCG-unresponsive non-muscle-invasive bladder cancer： interim results［J］. Urol Oncol， 2018， 36（10）： 440-447. DOI： 10.1016/j.urolonc.2017.07.005http://dx.doi.org/10.1016/j.urolonc.2017.07.005.

ZHANG S N， CHOI I K， HUANG J H， et al. Optimizing DC vaccination by combination with oncolytic adenovirus coexpressing IL-12 and GM-CSF［J］. Mol Ther， 2011， 19（8）： 1558-1568. DOI： 10.1038/mt.2011.29http://dx.doi.org/10.1038/mt.2011.29.

LEE Y S， KIM J H， CHOI K J， et al. Enhanced antitumor effect of oncolytic adenovirus expressing interleukin-12 and B7-1 in an immunocompetent murine model［J］. Clin Cancer Res， 2006， 12（19）： 5859-5868. DOI： 10.1158/1078-0432.CCR-06-0935http://dx.doi.org/10.1158/1078-0432.CCR-06-0935.

CHOI I K， LI Y， OH E， et al. Oncolytic adenovirus expressing IL-23 and p35 elicits IFN-γ- and TNF-α- co-producing T cell-mediated antitumor immunity［J］. PLoS One， 2013， 8（7）： e67512. DOI： 10.1371/journal.pone. 0067512http://dx.doi.org/10.1371/journal.pone.0067512.

YANG P H， SUN F， WANG R L， et al. Oncolytic activity of a novel influenza A virus carrying granulocyte-macrophage colony-stimulating factor in hepatocellular carcinoma［J］. Hum Gene Ther， 2019， 30（3）： 330-338. DOI： 10.1089/hum.2018.095http://dx.doi.org/10.1089/hum.2018.095.

YU H Y， SUN F， XU Y， et al. Combination immunotherapy of oncolytic flu-vectored virus and programmed cell death 1 blockade enhances antitumor activity in hepatocellular carcinoma［J］. Hum Gene Ther， 2024， 35（5/6）： 177-191. DOI： 10.1089/hum.2023.150http://dx.doi.org/10.1089/hum.2023.150.

LIANG C， SHI S， MENG Q C， et al. Complex roles of the stroma in the intrinsic resistance to gemcitabine in pancreatic cancer： where we are and where we are going［J］. Exp Mol Med， 2017， 49（12）： e406. DOI： 10.1038/emm. 2017.255http://dx.doi.org/10.1038/emm.2017.255.

JIANG B L， ZHOU L， LU J， et al. Stroma-targeting therapy in pancreatic cancer： one coin with two sides？［J］. Front Oncol， 2020， 10： 576399. DOI： 10.3389/fonc.2020. 576399http://dx.doi.org/10.3389/fonc.2020.576399.

SHALHOUT S Z， MILLER D M， EMERICK K S， et al. Therapy with oncolytic viruses： progress and challenges［J］. Nat Rev Clin Oncol， 2023， 20（3）： 160-177. DOI： 10.1038/s41571-022-00719-whttp://dx.doi.org/10.1038/s41571-022-00719-w.

POL J， KROEMER G， GALLUZZI L. First oncolytic virus approved for melanoma immunotherapy［J］. Oncoimmunology， 2016， 5（1）： e1115641. DOI： 10.1080/2162402X.2015.1115641http://dx.doi.org/10.1080/2162402X.2015.1115641.

LIU T C， HWANG T， PARK B H， et al. The targeted oncolytic poxvirus JX-594 demonstrates antitumoral， antivascular， and anti-HBV activities in patients with hepatocellular carcinoma［J］. Mol Ther， 2008， 16（9）： 1637-1642. DOI： 10.1038/mt.2008.143http://dx.doi.org/10.1038/mt.2008.143.

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. DOI： 10.1016/S1470-2045（08）70107-4http://dx.doi.org/10.1016/S1470-2045（08）70107-4.

THORNE S H， HWANG T H， O’GORMAN W E， et al. Rational strain selection and engineering creates a broad-spectrum， systemically effective oncolytic poxvirus， JX-963［J］. J Clin Invest， 2007， 117（11）： 3350-3358. DOI： 10.1172/JCI32727http://dx.doi.org/10.1172/JCI32727.

SANTOS APOLONIO J， LIMA DE SOUZA GONÇALVES V， CORDEIRO SANTOS M L， et al. Oncolytic virus therapy in cancer： a current review［J］. World J Virol， 2021， 10（5）： 229-255. DOI： 10.5501/wjv.v10.i5.229http://dx.doi.org/10.5501/wjv.v10.i5.229.

JARNAGIN W R， ZAGER J S， HEZEL M， et al. Treatment of cholangiocarcinoma with oncolytic herpes simplex virus combined with external beam radiation therapy［J］. Cancer Gene Ther， 2006， 13（3）： 326-334. DOI： 10.1038/sj.cgt.7700890http://dx.doi.org/10.1038/sj.cgt.7700890.

OHASHI K， KOBAYASHI G， FANG S， et al. Surgical excision combined with autologous whole tumor cell vaccination is an effective therapy for murine neuroblastoma［J］. J Pediatr Surg， 2006， 41（8）： 1361-1368. DOI： 10.1016/j.jpedsurg.2006.04.034http://dx.doi.org/10.1016/j.jpedsurg.2006.04.034.

CHU Y W， WANG L X， YANG G J， et al. Efficacy of GM-CSF-producing tumor vaccine after docetaxel chemotherapy in mice bearing established Lewis lung carcinoma［J］. J Immunother， 2006， 29（4）： 367-380. DOI： 10.1097/01.cji.0000199198.43587.bahttp://dx.doi.org/10.1097/01.cji.0000199198.43587.ba.

OH D Y， FONG L， NEWELL E W， et al. Toward a better understanding of T cells in cancer ［J］. Cancer Cell， 2021， 39（12）： 1549-1552. DOI： 10.1016/j.ccell.2021.11.010http://dx.doi.org/10.1016/j.ccell.2021.11.010.

FENG B， BAI Z， ZHOU X， et al. The type 2 cytokine Fc-IL-4 revitalizes exhausted CD8（+） T cells against cancer ［J］. Nature， 2024， 634（8034）： 712-720. DOI： 10.1038/s41586-024-07962-4http://dx.doi.org/10.1038/s41586-024-07962-4.

MACH N， GILLESSEN S， WILSON S B， et al. Differences in dendritic cells stimulated in vivo by tumors engineered to secrete granulocyte-macrophage colony-stimulating factor or Flt3-ligand［J］. Cancer Res， 2000， 60（12）： 3239-3246.

LIU S Y， LI F， MA Q Q， et al. OX40L-armed oncolytic virus boosts T-cell response and remodels tumor microenvironment for pancreatic cancer treatment［J］. Theranostics， 2023， 13（12）： 4016-4029. DOI： 10.7150/thno.83495http://dx.doi.org/10.7150/thno.83495.

WEI W， TIAN L Q， ZHENG X Y， et al. Expression of GPX4 by oncolytic vaccinia virus can significantly enhance CD8+T cell function and its impact against pancreatic ductal adenocarcinoma［J］. Oncoimmunology， 2024， 13（1）： 2322173. DOI： 10.1080/2162402X.2024.2322173http://dx.doi.org/10.1080/2162402X.2024.2322173.

ARAKI H， TAZAWA H， KANAYA N， et al. Oncolytic virus-mediated p53 overexpression promotes immunogenic cell death and efficacy of PD-1 blockade in pancreatic cancer［J］. Mol Ther Oncolytics， 2022， 27： 3-13. DOI： 10.1016/j.omto.2022.09.003http://dx.doi.org/10.1016/j.omto.2022.09.003.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effect of LncRNA LOC100506123 on proliferation and migration in pancreatic cancer cells

Expression of circRNA-IARS in pancreatic carcinoma and its clinical significance

ALDH1A3 regulates invasion ability of pancreatic cancer cells

Related Author

Rajiv Kumar JHA1

ZHANG Xudong1

LIU Hao’ang1

WANG Zhihao1

LAN Kai1

CHEN Rui2

LI Kai1

LIU Shuyi1

Related Institution

Department of Obstetrics and Gynecology, First Affiliated Hospital of Xi’an Medical University

Nepal Friendship Medical Research Center of Rajiv Kumar Jha, School of Clinical Medicine, Xi’an Medical University

重庆医科大学：附属第一医院泌尿外科，分子肿瘤及表观遗传学重庆市重点实验室

Institute of Hepatobiliary Surgery, Southwest Hospital, Institute of Immunology, College of Basic Medical Sciences, Third Military Medical University

Institute of Hepatobiliary Surgery, First Affiliated Hospital, Army Medical University (Third Military Medical University)

⁰

DOI：10.16016/j.2097-0927.202407052

摘要

Abstract

关键词

Keywords

references