Avastin联合抗原致敏的DC-CTL细胞体外抑制口腔鳞癌细胞生长的实验研究

doi:10.13701/j.cnki.kqyxyj.2016.08.018

口腔医学研究 ›› 2016, Vol. 32 ›› Issue (8): 848-852.DOI: 10.13701/j.cnki.kqyxyj.2016.08.018

Avastin联合抗原致敏的DC-CTL细胞体外抑制口腔鳞癌细胞生长的实验研究

蒋丽娟¹,郑阳²,泥艳红¹,韩伟¹,黄晓峰¹,胡勤刚¹,王志勇^1*

1. 南京大学附属口腔医院颌面外科江苏南京 210000;
2. 南京市同喆口腔门诊江苏南京 210000

收稿日期:2015-11-03 出版日期:2016-08-26 发布日期:2016-08-26
通讯作者: 王志勇,E-mail:wangzhiyong67@163.com
作者简介:蒋丽娟(1987~ ),女,江苏人,硕士,主要从事口腔颌面外科工作。
基金资助:
江苏省重点医学人才项目(RC2011025)

Inhibition of Cell Growth by Avastin Combined with CTLs in Oral Squamous Cell Carcinoma

JIANG Li-juan¹, ZHENG Yang², NI Yan-hong¹, HAN Wei¹, HUANG Xiao-feng¹, HU Qin-gang¹, WANG Zhi-yong

1. Affiliated Stomatological Hospital of Nanjing University, Nanjing 210000, China;
2. Tongzhe Dental Clinic, Nanjing 210000, China.

Received:2015-11-03 Online:2016-08-26 Published:2016-08-26

摘要/Abstract

摘要： 目的:研究贝伐单抗(Bevacizumab,Avastin)与口腔鳞癌抗原致敏的树突状细胞(Dendritic Cell,DC)诱导的细胞毒性T淋巴细胞(Cytotoxic t Lymphocyte,CTL)联合应用,体外研究其对口腔鳞癌细胞生长的抑制作用。方法:CAL27、SCC4肿瘤细胞系的培养,检测VEGF分泌水平;培养人外周血单核细胞来源的树突状细胞(DC);冻融法制备SCC4口腔鳞癌细胞可溶性抗原并致敏DC,检测DC表面标志物变化,刺激同种异体T细胞增殖的能力及IL-12分泌水平;将同源人T淋巴细胞与之共同培养诱导出抗原特异性的CTL细胞,Avastin联合CTL对SCC4细胞进行体外杀伤实验。结果:SCC4较CAL27分泌更高水平的VEGF;经肿瘤抗原致敏后的DC,其表面标志CD83、CD80、CD86、CD40及功能相关分子IL-12表达上调,CD1a的表达下调,可显著刺激异体T淋巴细胞增殖,具有统计学差异;Avastin联合CTL对SCC4的杀伤率达56.1%,显著高于对照组(P<0.05)。结论:Avastin联合DC致敏的CTL能显著抑制口腔癌细胞的生长。

关键词: 树突状细胞, 血管内皮生长因子, 贝伐单抗, 口腔鳞癌细胞

Abstract: Objective: To investigate the effect of anti-VEGF monoclonal antibody bevacizumab (Avastin) combined with CTLs induced by tumor antigen-sensitized dendritic cells on inhibiting the growth of oral squamous cell carcinoma (OSCC). Methods: VEGF production in the supernatants of cultured CAL27 and SCC4 were analyzed by enzyme-linked immunoabsorbent assay (ELISA). DCs were generated from human peripheral blood monocytes (PBMCs) and sensitized by SCC4 cell lysates. After that, their phenotypes were analyzed by flow cytometry (FCM), mixed lymphocyte reaction (MLR) for allostimulatory function and IL-12 level was detected by ELISA. DCs loaded with tumor antigen were co-cultured with allogeneic T cells to induce tumor specific cytotoxic T lymphocytes (CTLs). Meanwhile, killing effect of Avastin combined with CTLs to SCC4 in vitro was studied. Results: The level of VEGF secreted by SCC4 was higher than CAL27. Compared with the control group, the expression of CD83, HLA-DR, CD86, CD80 and CD40 on DCs loaded with SCC4 cell lysate, the level of IL-12 and the stimulatory capacity to T cell proliferation were obviously enhanced, but the expression of CD1a was down-regulated. DCs loaded with OSCC antigen increased the proliferation of T lymphocytes. The killing effect of Avastin combined with CTLs was 56.1% to SCC4 and was significantly higher than that of control group (P<0.05). Conclusion: Avastin combined with CTLs induced by tumor-antigen sensitized DCs could inhibit OSCC growth.

Key words: Dendritic cell, VEGF, Bevacizumab, Oral squamous carcinoma cells

中图分类号:

R739.8

蒋丽娟,郑阳,泥艳红,韩伟,黄晓峰,胡勤刚,王志勇. Avastin联合抗原致敏的DC-CTL细胞体外抑制口腔鳞癌细胞生长的实验研究[J]. 口腔医学研究, 2016, 32(8): 848-852.

JIANG Li-juan, ZHENG Yang, NI Yan-hong, HAN Wei, HUANG Xiao-feng, HU Qin-gang, WANG Zhi-yong. Inhibition of Cell Growth by Avastin Combined with CTLs in Oral Squamous Cell Carcinoma[J]. Journal of Oral Science Research, 2016, 32(8): 848-852.

参考文献

[1] 郑家伟,李金忠,钟来平,等.口腔鳞状细胞癌临床流行病学研究现状[J].中国口腔颌面外科, 2007, 5(2)∶83-90
[2] Galluzzi L, Senovilla L, Vacchelli E, et al. trial watch: Dendritic cell-based interventions for cancer therapy [J]. Oncoimmunology, 2012, 1(7)∶1111-1116
[3] Gabrilovich DI, Nadaf S, Corak J, et al. Dendritic cells in antitumor responses. II. Dendritic cells grown from bone marrow precursor, but not mature DC form tumor-bearing mice, are effective antigen carriers in the therapy of established tumors [J]. Cell Immunol,1996,170(1)∶111-119
[4] Villaruz LC, Socinski MA. The role of anti-angiogenesis in non-small-cell lung cancer: an update [J]. Curr Oncol Rep, 2015, 17(6)∶448-451
[5] Cameron D,. Adjuvant bevacizumab-containing therapy in triple-negative breast cancer (BEATRICE): Primary results of a randomised, phase 3 trial [J]. Lancet Oncol, 2013, 14(10)∶933-942
[6] Zhai H, Zhong W, Yang X, et al. Neoadjuvant and adjuvant epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) therapy for lung cancer [J]. Transl Lung Cancer Res, 2015, 4(1)∶82-93
[7] Flaherty KT. Inhibition of mutated, activated BRAF in metastatic melanoma [J]. N. Engl. J. Med,2010,363(5)∶809-819
[8] Redman JM, Gibney GT, Atkins MB. Advances in immunotherapy for melanoma [J]. BMC medicine, 2016, 14(1)∶20-24
[9] Cimino-Mathews A., Foote J. B., Emens L. A. Immune targeting in breast cancer [J]. Oncology, 2015,29(2)∶375-385
[10] Wang ZY, Shi PH, Huang XF, et al. Peripheral blood dendritic cells and vascular endothelial growth factor in oral squamous cell carcinoma: correlation analysis and in vitro study [J]. Int J Oral Maxillofac Surg, 2010,39(5)∶713-720
[11] Ni YH, Wang ZY, Huang XF, et al. Effect of siRNA-mediated downregulation of VEGF in tca8113 cells on the activity of monocyte-derived dendritic cells [J]. Oncol Lett, 2012,3(6)∶885-892
[12] Su, H. therapeutic antitumor efficacy of tumor-derived autophagosome (DRibble) vaccine on head and neck cancer [J]. Int J Nanomedicine, 2015, 31(9)∶1921-30
[13] Chen Z, Moyanat, Saxena A, et al. Efficient antitumor immunity derived from maturation of dendritic cells that had phagocytosed apoptotic/necrotic tumor cells [J]. International journal of cancer, 2001, 93(4)∶539-548
[14] tahara H, Lotze M t. Antitumor effects of interleukin-12 (IL-12): applications for the immunotherapy and gene therapy of cancer [J]. Gene therapy, 1995, 2(2)∶96-106
[15] Oyamat, Ran S, Ishida t, et al. Vascular endothelial growth factor affects dendritic cell maturation through the inhibition of nuclear factor-κB activation in hemopoietic progenitor cells [J]. The Journal of Immunology, 1998, 160(3)∶1224-123
[16] Mimura K, Kono K, takahashi A, et al. Vascular endothelial growth factor inhibits the function of human mature dendritic cells mediated by VEGF receptor-2 [J]. Cancer ImmunolImmunother, 2007, 56(6)∶761-70

Avastin联合抗原致敏的DC-CTL细胞体外抑制口腔鳞癌细胞生长的实验研究

Inhibition of Cell Growth by Avastin Combined with CTLs in Oral Squamous Cell Carcinoma

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