PO.TB02.01 · 肿瘤生物学
在细胞来源异种移植小鼠模型中使用抗 mucin-4 抗体偶联近红外荧光团靶向原发性胃肿瘤和腹膜转移灶
Targeting primary gastric tumors and peritoneal metastases in cell-derived xenograft mouse models using an anti-mucin-4-antibody conjugated near infrared fluorophore
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
引言：本研究的目的是验证使用抗 MUC4 抗体偶联的近红外染料在裸鼠模型中靶向原发性和转移性胃癌的应用。
方法：通过将 10^6 个人胃癌细胞系 SNU-16 或 NCI-N87 细胞注射到裸鼠胁部，建立皮下模型。这些肿瘤在 2 个月后被切除，并将 1 mm³ 的碎片植入裸鼠的胃中，以建立 SNU-16 和 NCI-N87 的原位胃癌模型。通过在裸鼠腹腔内注射 10^6 个 SNU-16 或 10^6 个 NCI-N87 细胞建立实验性转移模型。肿瘤植入 2-3 个月后，经尾静脉向小鼠注射偶联近红外荧光染料 IRDye800CW 的 mucin4 抗体（MUC4-IRDye800CW）。在尾静脉注射 50 μg MUC4-IRDye800CW 后 72 小时处死小鼠，并使用 LI-COR Pearl 成像系统和 Stryker 腹腔镜进行成像。通过将肿瘤的平均荧光强度除以周围胃组织的平均荧光强度来计算肿瘤-背景比（TBR）。对肿瘤进行组织病理学分析，采用抗 mucin-4 和抗免疫球蛋白 G（IgG）（作为阴性对照）进行免疫组织化学（IHC）染色。
结果：共对 20 只小鼠进行了成像，包括 14 个原发性胃癌模型和 4 个腹腔内转移性胃癌模型。原发性胃 SNU-16 肿瘤（n = 6）的平均 TBR 为 1.99（SE ± 0.58），原发性胃 NCI-N87 肿瘤（n = 8）的平均 TBR 为 2.81（SE ± 1.04）。转移性 SNU-16 肿瘤（n = 3）的平均 TBR 为 3.7（SE ± 0.75），转移性 NCI-N87 肿瘤（n = 3）的平均 TBR 为 3.91（SE ± 0.98）。使用 Pearl 和 Stryker 腹腔镜荧光成像系统均能清晰显示肿瘤边界。经抗 mucin-4 染色的代表性肿瘤呈棕色，而经抗 IgG 染色的肿瘤则不着色。
结论：抗 MUC4-IRDye800CW 可用于通过荧光成像准确定位裸鼠中的原发性和转移性胃癌模型，并有望在术中识别原发肿瘤边界和腹腔内转移灶。
查看英文原文 English abstract
Introduction: The purpose of the present study is to validate the use of an anti-MUC4 antibody conjugated near infrared dye to target primary and metastatic gastric cancers in nude mouse models.
Methods: Subcutaneous models were created in nude mice by injecting 10 6 cells of human gastric cancer cell lines SNU-16 or NCI-N87 into their flanks. These tumors were excised after 2 months and 1 mm 3 fragments were implanted into the stomachs of nude mice to establish orthotopic gastric cancer models of SNU-16 and NCI-N87. Experimental metastasis models were established by injecting 10 6 SNU-16 or 10 6 NCI-N87 cells intraperitoneally in nude mice. 2-3 months after tumor implantation mice were injected via tail vein with a mucin4 antibody conjugated to a near infrared fluorescent dye IRDye800CW (MUC4-IRDye800CW). Mice were sacrificed 72 hours after tail vein injection of 50 µg MUC4-IRDye800CW and imaged with the LI-COR Pearl Imaging System and a Stryker laparoscope. The tumor-to-background ratios (TBR) were calculated by dividing the mean fluorescence intensity of the tumors by that of the surrounding stomach. Histopathological analysis was performed on tumors with immunohistochemical (IHC) staining with anti-mucin-4 and anti-immunoglobulin G (IgG) as a negative control.
Results: 20 total mice were imaged, including 14 primary gastric cancer models and 4 intraperitoneal metastatic gastric cancer models. Primary gastric SNU-16 tumors (n = 6) had a mean TBR of 1.99 (SE ± 0.58), and primary gastric NCI-N87 tumors (n = 8) had a mean TBR of 2.81 (SE ± 1.04). Metastatic SNU-16 tumors (n = 3) had a mean TBR of 3.7 (SE ± 0.75), and metastatic NCI-N87 tumors (n = 3) had a mean TBR of 3.91 (SE ± 0.98). Tumor borders were clearly visualized with both Pearl and the Stryker laparoscope fluorescence imaging systems. Representative tumors stained with anti-mucin-4 stained brown while those stained with anti-IgG did not.
Conclusions: Anti-MUC4-IRDye800CW can be used to accurately localize both primary and metastatic gastric cancer models in nude mice with fluorescence imaging and is promising for identifying both primary tumor margins and intraperitoneal metastasis intra-operatively.
利益披露 Disclosure
B. Eddins, None..
S. Jaiswal, None..
A. Aithal, None..
S. Amirfakhri, None..
J. Bravo, None..
J. Zaker, None..
S. Talib, None..
K. B. Mallya, None..
A. M. Mohs, None..
M. Jain, None..
S. K. Batra, None..
M. Bouvet, None.