Immunotherapy for hepatoma using a dual-function vector with both immunostimulatory and pim-3-silencing effects.
Tumorigenesis is an immortalization process in which the growth of normal cells is uncontrolled and programmed cell death is suppressed. Molecular biologic and immunologic studies have revealed that the aberrant expression of some proto-oncogenes boosts proliferation and inhibits apoptosis, which is vital for tumor development. The hypofunction of the host immune system also drives the development and metastasis of malignant tumors. Pim-3, a member of the Pim family, is aberrantly expressed in several cancers. Data suggest that Pim-3 inhibits apoptosis by phosphorylating the proapoptotic BH3-only protein Bad. Here, we constructed a dual-function small hairpin RNA (shRNA) vector containing an shRNA targeting Pim-3 and a TLR7-stimulating ssRNA. Stimulation with this bi-functional vector in vitro promoted significant apoptosis of Hepa1-6 cells by regulating the expression of apoptosis-related proteins and induced secretion of type I IFNs. Most importantly, this bi-functional vector more effectively inhibited subcutaneous Hepa1-6 cell growth than did single shRNA and ssRNA treatment in vivo. Natural killer (NK), CD4(+) T, and CD8(+) T cells and macrophages were required for effective tumor suppression, and CD4(+) T cells were shown to play a helper role in the activation of NK cells, possibly by regulating the secretion of Th1 or Th2 cytokines. This ssRNA-shRNA bi-functional vector may represent a promising approach for tumor therapy.
Authors' Affiliations: Institute of Immunopharmacology & Immunotherapy, School of Pharmaceutical Sciences, Shandong University, Jinan; Department of Pharmacy, The Affiliated Hospital of Medical College, Qingdao University, Qingdao, Shandong; and Department of Microbiology and Immunology, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, ChinaAuthors' Affiliations: Institute of Immunopharmacology & Immunotherapy, School of Pharmaceutical Sciences, Shandong University, Jinan; Department of Pharmacy, The Affiliated Hospital of Medical College, Qingdao University, Qingdao, Shandong; and Department of Microbiology and Immunology, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, China.
Qie
Q
Guo
Authors' Affiliations: Institute of Immunopharmacology & Immunotherapy, School of Pharmaceutical Sciences, Shandong University, Jinan; Department of Pharmacy, The Affiliated Hospital of Medical College, Qingdao University, Qingdao, Shandong; and Department of Microbiology and Immunology, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, China.
Peixiang
P
Lan
Authors' Affiliations: Institute of Immunopharmacology & Immunotherapy, School of Pharmaceutical Sciences, Shandong University, Jinan; Department of Pharmacy, The Affiliated Hospital of Medical College, Qingdao University, Qingdao, Shandong; and Department of Microbiology and Immunology, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, China.
Xin
X
Yu
Authors' Affiliations: Institute of Immunopharmacology & Immunotherapy, School of Pharmaceutical Sciences, Shandong University, Jinan; Department of Pharmacy, The Affiliated Hospital of Medical College, Qingdao University, Qingdao, Shandong; and Department of Microbiology and Immunology, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, China.
Qiuju
Q
Han
Authors' Affiliations: Institute of Immunopharmacology & Immunotherapy, School of Pharmaceutical Sciences, Shandong University, Jinan; Department of Pharmacy, The Affiliated Hospital of Medical College, Qingdao University, Qingdao, Shandong; and Department of Microbiology and Immunology, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, China.
Jian
J
Zhang
Authors' Affiliations: Institute of Immunopharmacology & Immunotherapy, School of Pharmaceutical Sciences, Shandong University, Jinan; Department of Pharmacy, The Affiliated Hospital of Medical College, Qingdao University, Qingdao, Shandong; and Department of Microbiology and Immunology, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, ChinaAuthors' Affiliations: Institute of Immunopharmacology & Immunotherapy, School of Pharmaceutical Sciences, Shandong University, Jinan; Department of Pharmacy, The Affiliated Hospital of Medical College, Qingdao University, Qingdao, Shandong; and Department of Microbiology and Immunology, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, China.
Zhigang
Z
Tian
Authors' Affiliations: Institute of Immunopharmacology & Immunotherapy, School of Pharmaceutical Sciences, Shandong University, Jinan; Department of Pharmacy, The Affiliated Hospital of Medical College, Qingdao University, Qingdao, Shandong; and Department of Microbiology and Immunology, School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, China caizhangsd@sdu.edu.cn.
Cai
C
Zhang
D017209:Apoptosis / Q000235:genetics; D063646:Carcinogenesis / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D020868:Gene Silencing; D005822:Genetic Vectors; D006801:Humans; D007167:Immunotherapy; D008113:Liver Neoplasms / Q000235:genetics / Q000276:immunology / Q000628:therapy*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics
D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; C499249:PIM3 protein, human; D017346:Protein Serine-Threonine Kinases