title: Anticancer effects of adenovirus-mediated calreticulin and melanoma-associated antigen 3 expression on non-small cell lung cancer cells.
abstract: Non-small cell lung cancer (NSCLC) is highly prevalent and needs novel therapies. Melanoma-associated antigen 3 (MAGE-A3) is a lung cancer antigen and calreticulin (CALR) can modulate immune responses. Our previous study has shown that up-regulated MAGE-A3 and CALR expression inhibits the proliferation and invasion of glioma cells. In this study, we examined the effect of adenovirus (Ad)-mediated MAGE-A3 and/or CALR expression on the proliferation, invasion, and apoptosis of human NSCLC cells and on the vascular tube formation of human endothelial cells as well as on dendritic cell (DC) activation and induced CD8(+) cytotoxic T lymphocyte (CTL) activity in vitro. We found that low levels of CALR and MAGE-A3 were expressed by A549 cells, but only very low CALR was expressed by DC. Up-regulated CALR and MAGE-A3 expression by infection with Ad-CALR/MAGE-A3 significantly inhibited the proliferation and invasion, but promoted the apoptosis of A549 cells. Up-regulated CALR and MAGE-A3 expression significantly inhibited cyclin D1 expression and the AKT, ERK1/2 and NF-κB expression and phosphorylation in A549 cells. Up-regulated CALR expression inhibited the tube formation in human endothelial cells. Up-regulated CALR and MAGE-A3 expression synergistically enhanced classical DC activation by enhancing IL-12, but reducing IL-10 secretion. Furthermore, CTLs induced by up-regulated CALR and MAGE-A3 expressing DCs synergistically triggered A549 cell apoptosis, which was abrogated by treatment with anti-HLA I, but not anti-HLA II antibodies. Moreover, CTLs induced by CALR and MAGE-A3-expressing DCs had a higher frequency of A549-specific IFN-γ-secreting T cells. Our data indicated that up-regulated CALR and MAGE-A3 expression inhibited the carcinogenesis of NSCLC by modulating the AKT, ERK MAPK and NF-κB signaling and enhanced classical DC activation and MAGE-A3-specific CTL cytotoxicity. Therefore, our findings may provide new insights in understanding the role of CALR in modulating antigen-specific T cell immunity and may aid in the design of new therapies for NSCLC.
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China; Department of Medical Oncology, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: cinlylau@hotmail.com.
authors.forename: Xinli
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: nanan1997s@hotmail.com.
authors.forename: Nan
authors.initials: N
authors.lastname: Sun
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: dongyinan_2012@163.com.
authors.forename: Yinan
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: lijijia1982@163.com.
authors.forename: Jijia
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Medical Oncology, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: lyylegend@hotmail.com.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: renyi1620@sina.com.
authors.forename: Yi
authors.initials: Y
authors.lastname: Ren
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: yangliang_0927@163.com.
authors.forename: Chengliang
authors.initials: C
authors.lastname: Yang
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: zl750523@sina.com.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: janezy1987@163.com.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: 910397562@qq.com.
authors.forename: Zhuang
authors.initials: Z
authors.lastname: Tong
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: djq1988@126.com.
authors.forename: Jianqiao
authors.initials: J
authors.lastname: Ding
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: zhuningzhu13@126.com.
authors.forename: Xiang
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: liuyu1984@126.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: 63788321@qq.com.
authors.forename: Rong
authors.initials: R
authors.lastname: He
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: 1776119@qq.com.
authors.forename: Ke
authors.initials: K
authors.lastname: Xu
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: doc.wuyongkai@gmail.com.
authors.forename: Yongkai
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: huangyanfeng227@163.com.
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of Medical Oncology, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: 550971948@qq.com.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Medical Oncology, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: liuyang2020@hotmail.com.
authors.forename: Xiujuan
authors.initials: X
authors.lastname: Tao
authors.affiliation: Department of Thoracic Surgery, Liaoning Provincial Tumor Hospital, Shenyang 110042, China. Electronic address: lyymot@hotmail.com.
authors.forename: Yongyu
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000256:Adenoviridae; D000951:Antigens, Neoplasm / Q000378:metabolism*; D017209:Apoptosis; D037282:Calreticulin / Q000378:metabolism*; D002289:Carcinoma, Non-Small-Cell Lung / Q000378:metabolism*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000502:physiology; D006801:Humans; D016753:Interleukin-10 / Q000378:metabolism; D018664:Interleukin-12 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D000951:Antigens, Neoplasm; C576483:CALR protein, human; D037282:Calreticulin; C508609:IL10 protein, human; C072013:MAGEA3 protein, human; D016328:NF-kappa B; D009363:Neoplasm Proteins; D016753:Interleukin-10; D018664:Interleukin-12; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: CD8+ T cells; Calreticulin; Dendritic cells; Melanoma antigen gene-A3; Non-small cell lung cancer