成像胰腺癌与肿瘤微环境中星状细胞在转移过程中的相互作用。

Imaging the Interaction of Pancreatic Cancer and Stellate Cells in the Tumor Microenvironment during Metastasis.

作者信息

Suetsugu Atsushi, Snyder Cynthia S, Moriwaki Hisataka, Saji Shigetoyo, Bouvet Michael, Hoffman Robert M

机构信息

AntiCancer, Inc., San Diego, CA, U.S.A. Department of Surgery, University of California, San Diego, CA, U.S.A. Gifu University Graduate School of Medicine, Gifu, Japan.

Department of Surgery, University of California, San Diego, CA, U.S.A.

出版信息

Anticancer Res. 2015 May;35(5):2545-51.

PMID:25964528

Abstract

BACKGROUND/AIM: Pancreatic stellate cells are involved in fibrosis of pancreatic cancer termed desmoplasia, which may contribute to both pancreatic cancer growth and metastasis, as well as to drug resistance. A better understanding of pancreatic cancer-cell interactions with stellate cells is therefore critical to our ability to develop effective anti-metastatic therapeutics for pancreatic cancer.

MATERIALS AND METHODS

The human pancreatic cancer cell line XPA-1 was engineered to express green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in the cytoplasm. Pancreatic stellate cells were engineered to express RFP. The pancreatic cancer cells and stellate cells were co-cultured and their interaction was imaged in vitro. The pancreatic cancer cells and stellate cells were then co-injected in the spleen of transgenic cyan fluorescent protein (CFP) nude mice and imaged in liver, lung and diaphragm metastasis.

RESULTS

The interaction of the pancreatic cancer cells expressing GFP in the nucleus and RFP in the cytoplasm and stellate cells expressing RFP was first imaged in vitro. The intimate relationship between the two cell types could be seen. Three hours after splenic co-injection, dual-color pancreatic cancer cells and pancreatic stellate cells were found distributed in the host liver. By 28 days after splenic co-injection of the pancreatic cancer and stellate cells, liver metastases were observed in host CFP nude mice. Metastases were also observed in the lung and diaphragm. Stellate cells were observed along with the pancreatic cancer cells at all metastatic sites suggesting that stellate cells may be necessary for metastasis. With high-resolution intravital imaging afforded by the Olympus FV1000 confocal microscope, the interaction of the dual-colored pancreatic cancer cells and the RFP-expressing pancreatic stellate cells could be clearly imaged in the liver and other metastases, further suggesting that stellate cells participate in metastasis formation.

CONCLUSION

Pancreatic cancer cells and stellate stem cells form a very close relationship and accompany each other to distant metastatic sties. Our hypothesis is that pancreatic stellate cells form a niche for metastasis of pancreatic cancer.

摘要

背景/目的：胰腺星状细胞参与胰腺癌的纤维化过程，即所谓的促纤维增生反应，这可能促进胰腺癌的生长、转移以及耐药性。因此，更好地理解胰腺癌细胞与星状细胞之间的相互作用对于我们开发有效的胰腺癌抗转移治疗方法的能力至关重要。

材料与方法

对人胰腺癌细胞系XPA-1进行基因改造，使其细胞核表达绿色荧光蛋白（GFP），细胞质表达红色荧光蛋白（RFP）。对胰腺星状细胞进行基因改造，使其表达RFP。将胰腺癌细胞和星状细胞共培养，并在体外对它们的相互作用进行成像。然后将胰腺癌细胞和星状细胞共同注射到转基因青色荧光蛋白（CFP）裸鼠的脾脏中，并对肝脏、肺和膈肌转移情况进行成像。

结果

首先在体外对细胞核表达GFP、细胞质表达RFP的胰腺癌细胞与表达RFP的星状细胞之间的相互作用进行了成像。可以看到这两种细胞类型之间的密切关系。脾脏共同注射3小时后，在宿主肝脏中发现了双色胰腺癌细胞和胰腺星状细胞。在脾脏共同注射胰腺癌和星状细胞28天后，在宿主CFP裸鼠中观察到肝脏转移。在肺和膈肌中也观察到转移。在所有转移部位都观察到星状细胞与胰腺癌细胞在一起，这表明星状细胞可能是转移所必需的。利用奥林巴斯FV1000共聚焦显微镜提供的高分辨率活体成像技术，可以在肝脏和其他转移灶中清晰地对双色胰腺癌细胞与表达RFP的胰腺星状细胞之间的相互作用进行成像，进一步表明星状细胞参与转移形成。