後根神経節モデル：癌性神経浸潤のin vitroモデルでは
Summary
This video article shows the use of the dorsal root ganglia (DRG)/cancer cell model in pancreatic ductal adenocarcinoma.
Abstract
One way that solid tumors disseminate is through neural invasion. This route is well-known in cancers of the head and neck, prostate, and pancreas. These neurotropic cancer cells have a unique ability to migrate unidirectionally along nerves towards the central nervous system (CNS). The dorsal root ganglia (DRG)/cancer cell model is a three dimensional (3D) in vitro model frequently used for studying the interaction between neural stroma and cancer cells. In this model, mouse or human cancer cell lines are grown in ECM adjacent to preparations of freshly dissociated cultured DRG. In this article, the DRG isolation protocol from mice, and implantation in petri dishes for co-culturing with pancreatic cancer cells are demonstrated. Five days after implantation, the cancer cells made contact with the DRG neurites. Later, these cells formed bridgeheads to facilitate more extensive polarized, neurotropic migration of cancer cells.
Introduction
直接浸潤、リンパ節転移、および造血スプレッド：固形腫瘍は、主に次の3通りの方法で広めます。しかし、頻繁に神経に沿って、普及を無視して、がんの広がりの第4の手段があります。癌性神経浸潤（CNI）は、特に頭頸部の癌において、前立腺1 2、3及び膵臓癌の広がりの周知の経路である。4-8 CNIは、先頭、膵臓腺癌を有する個体の80％以上で起こります後腹膜腫瘍を腹腔神経節神経を介して広がります。これらの神経向性癌細胞は、中枢神経系（CNS）に向かって神経に沿って一方向に移行するためのユニークな能力を有する。9この知見は、神経周囲の微小環境は、腫瘍細胞によって利用され得ることを示唆している悪性の増殖を支持する因子を提供します。
CNIの研究のためのいくつかのin vitroモデルの一つは、後根神経節（DRG）/癌細胞モデルです。このモッズELが頻繁に神経間質細胞と癌細胞との間のパラクリン相互作用を研究するために使用される。10-18このモデルでは、マウスまたはヒトの癌細胞株は、新たに解離し、培養DRG調製物に隣接する細胞外マトリックス（ECM）中で増殖させます。
このビデオの記事では、膵管腺癌におけるインビトロCNIのアプリケーションを示しています。
Protocol
Representative Results
Discussion
この記事では、神経ニッチ、DRGモデルにおける癌微小環境を再現するin vitroモデルを提示します。ビデオは、マウスにおけるDRG、その抽出、およびECMで最終的に、その培養として、解剖学的ランドマークを認識することから始めて、すべての手順を示しています。共培養癌細胞と一緒にDRGにも提示されています。 in vitroでの神経周囲ニッチ微小環境を研究するためのこのモデル?…
Offenlegungen
The authors have nothing to disclose.
Acknowledgements
Edith Suss-Toby is thanked for her assistance in the time-lapse microscopy and image analysis. Nofar Rada is thanked for the artistic work.
Materials
| Equipments: | |||
| Operating microscope | Leica | M205 | |
| Tiime Lapse System | Zeiss | ||
| Forceps | Sigma-Aldrich | F4142 | |
| Surgical blade | Sigma-Aldrich | Z309036 | |
| Scissors | Sigma-Aldrich | S3271 | |
| 35mm petri dishes, glass bottom | de groot | 60-627860 | |
| Name | Company | Catalog Number | Comments |
| Materials: | |||
| 70% ethanol | sigma | ||
| Cold PBS | Biological industries | 02-023-1A | |
| DMEM | Biological industries | 01-055-1A | |
| FCS | Rhenium | 10108165 | |
| Penicillin and streptomycin | Biological industries | 01-031-1B | |
| Sodium Pyruvate | Biological industries | 03-042-1B | |
| L-Glutamine | Biological industries | 03-020-1B | |
| Growth factor depleted matrigel | Trevigen | 3433-005-01 |
Referenzen
- Carter, R. L., Foster, C. S., Dinsdale, E. A., Pittam, M. R. Perineural spread by squamous carcinomas of the head and neck, a morphological study using antiaxonal and antimyelin monoclonal antibodies. J Clin Pathol. 36, 269-275 (1983).
- Beard, C. J., et al. Perineural invasion is associated with increased relapse after external beam radiotherapy for men with low-risk prostate cancer and may be a marker for occult, high-grade cancer. Int J Radiat Oncol Biol Phys. 58, 19-24 (2004).
- Maru, N., Ohori, M., Kattan, M. W., Scardino, P. T., Wheeler, T. M. Prognostic significance of the diameter of perineural invasion in radical prostatectomy specimens. Hum Pathol. 32, 828-833 (2001).
- Ceyhan, G. O., et al. Pancreatic neuropathy and neuropathic pain–a comprehensive pathomorphological study of 546 cases. Gastroenterology. 136, 177-186 (2009).
- Ceyhan, G. O., et al. The neurotrophic factor artemin promotes pancreatic cancer invasion. Ann Surg. 244, 274-281 (2006).
- Takahashi, T., et al. Perineural invasion by ductal adenocarcinoma of the pancreas. J Surg Oncol. 65, 164-170 (1997).
- Zhu, Z., et al. Nerve growth factor expression correlates with perineural invasion and pain in human pancreatic cancer. J Clin Oncol. 17, 2419-2428 (1999).
- Hirai, I., et al. Perineural invasion in pancreatic cancer. Pancreas. 24, 15-25 (2005).
- Mitchem, J. B., et al. Targeting tumor-infiltrating macrophages decreases tumor-initiating cells, relieves immunosuppression, and improves chemotherapeutic responses. Cancer Res. 73, 1128-1141 (2013).
- Kelly, K., et al. Attenuated multimutated herpes simplex virus-1 effectively treats prostate carcinomas with neural invasion while preserving nerve function. FASEB J. 22, 1839-1848 (2008).
- Dai, H., et al. Enhanced survival in perineural invasion of pancreatic cancer, an in vitro approach. Hum Pathol. 38, 299-307 (2007).
- Ayala, G. E., et al. Cancer-related axonogenesis and neurogenesis in prostate cancer. Clin Cancer Res. 14, 7593-7603 (2008).
- Ayala, G. E., et al. Stromal antiapoptotic paracrine loop in perineural invasion of prostatic carcinoma. Cancer Res. 66, 5159-5164 (2006).
- Ceyhan, G. O., et al. Neural invasion in pancreatic cancer, a mutual tropism between neurons and cancer cells. Biochem Biophys Res Commun. 374, 442-447 (2008).
- Bapat, A. A., Hostetter, G., Von Hoff, D. D., Han, H. Perineural invasion and associated pain in pancreatic cancer. Nat Rev Cancer. 11, 695-707 (2011).
- Ketterer, K., et al. Reverse transcription-PCR analysis of laser-captured cells points to potential paracrine and autocrine actions of neurotrophins in pancreatic cancer. Clin Cancer Res. 9, 5127-5136 (2003).
- Gil, Z., et al. Nerve-sparing therapy with oncolytic herpes virus for cancers with neural invasion. Clin Cancer Res. 13, 6479-6485 (2007).
- Gil, Z., et al. Paracrine regulation of pancreatic cancer cell invasion by peripheral nerves. J Natl Cancer Inst. 102, 107-118 (2010).
- Weizman, N., et al. Macrophages mediate gemcitabine resistance of pancreatic adenocarcinoma by upregulating cytidinedeaminase. Oncogene. 33, 3812-3819 (2014).
