肿瘤密封外科原位植入人结肠癌裸鼠可诱导具有临床相关性的转移而无早期腹膜癌病。

Tumor-sealing Surgical Orthotopic Implantation of Human Colon Cancer in Nude Mice Induces Clinically-relevant Metastases Without Early Peritoneal Carcinomatosis.

机构信息

AntiCancer, Inc., San Diego, CA, U.S.A.

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

出版信息

Anticancer Res. 2019 Aug;39(8):4065-4071. doi: 10.21873/anticanres.13563.

DOI:10.21873/anticanres.13563

PMID:31366489

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11058567/

Abstract

BACKGROUND

Surgical orthotopic implantation of human colon cancer tissue to the ceca of mice has been used to mimic behavior of cancer in human patients for the development of precision cancer medicine. However, with the current method of serosal surface implantation (SSI) of pieces of human colon cancer tissue, cancer cells are exposed to the peritoneum, which can artificially increase the rate of peritoneal carcinomatosis (PC) during the disease course. The objective of the present study was to introduce a tumor-sealing method (TSM) and compare it with SSI for the ability to produce clinically-relevant metastases without artificial PC.

MATERIALS AND METHODS

HCT116 colon cancer cells transfected with green fluorescence protein (GFP) were cultured and then injected into the subcutaneous layer of athymic nude mice. Subcutaneous tumors were allowed to grow sufficiently to supply adequate tumor for orthotopic implantation. For SSI, a 1 mm-sized tumor fragment was sutured to partially torn serosa of the cecum. For TSM, the blind end of the cecum was folded over the tumor fragment and sealed with sutures. At 20 days after implantation, all mice were opened to visualize PC by intravital fluorescence imaging. At necropsy, distant metastasis was investigated using frozen section of whole blocks of organs.

RESULTS

At 20 days after implantation, PC rates in the SSI group and the TSM group were 80% (12/15) and 20% (3/15), respectively (p<0.001). The liver metastasis rate was 41.7% (5/12) in the SSI group and 50% (5/10) in the TSM group (p=0.696). The lung metastasis rate was 0% (0/12) in the SSI group and 10% (1/10) in the TSM group (p=0.201). The mean survival of mice without PC on the 20th day was significantly longer than that of mice with PC on the 20th day (69.1±14.7 vs. 44.5±12.4 days, p=0.001).

CONCLUSION

These results suggest that TSM might be a more patient-like and useful method as a model of metastatic colon cancer than SSI.

摘要

背景

将人结肠癌组织通过外科手术原位植入到小鼠的盲肠中，已被用于模拟人类患者的癌症行为，以开发精准癌症药物。然而，目前采用的浆膜表面植入（SSI）方法将人结肠癌组织块植入，使癌细胞暴露于腹膜中，这会人为增加疾病过程中的腹膜癌转移（PC）发生率。本研究的目的是引入一种肿瘤密封方法（TSM），并将其与 SSI 进行比较，以评估其在不产生人为 PC 的情况下产生临床相关转移的能力。

材料和方法

转染绿色荧光蛋白（GFP）的 HCT116 结肠癌细胞被培养，然后注入无胸腺裸鼠的皮下层。待皮下肿瘤生长到足以提供足够的肿瘤用于原位植入时，将其用于实验。对于 SSI，将 1mm 大小的肿瘤块缝合到部分撕裂的盲肠浆膜上。对于 TSM，将盲肠的盲端折叠在肿瘤块上并用缝线密封。在植入后 20 天，所有小鼠均通过活体荧光成像打开以可视化 PC。尸检时，使用器官全块的冷冻切片检查远处转移。

结果

在植入后 20 天，SSI 组和 TSM 组的 PC 发生率分别为 80%（12/15）和 20%（3/15）（p<0.001）。SSI 组的肝转移率为 41.7%（5/12），TSM 组为 50%（5/10）（p=0.696）。SSI 组的肺转移率为 0%（0/12），TSM 组为 10%（1/10）（p=0.201）。在第 20 天没有 PC 的小鼠的平均存活时间明显长于有 PC 的小鼠（69.1±14.7 与 44.5±12.4 天，p=0.001）。