在小鼠中用双顺反子腺病毒进行基因转移后，使用生长抑素受体和胸苷激酶进行γ相机双成像。

Gamma camera dual imaging with a somatostatin receptor and thymidine kinase after gene transfer with a bicistronic adenovirus in mice.

作者信息

Zinn Kurt R, Chaudhuri Tandra R, Krasnykh Victor N, Buchsbaum Donald J, Belousova Natalya, Grizzle William E, Curiel David T, Rogers Buck E

机构信息

Depts of Radiology, University of Alabama at Birmingham, Boshell Bldg, BDB 11, 1530 3rd Ave S, Birmingham, AL 35294-0012, USA.

出版信息

Radiology. 2002 May;223(2):417-25. doi: 10.1148/radiol.2232010501.

DOI:10.1148/radiol.2232010501

PMID:11997547

Abstract

PURPOSE

To compare two systems for assessing gene transfer to cancer cells and xenograft tumors with noninvasive gamma camera imaging.

MATERIALS AND METHODS

A replication-incompetent adenovirus encoding the human type 2 somatostatin receptor (hSSTr2) and the herpes simplex virus thymidine kinase (TK) enzyme (Ad-hSSTr2-TK) was constructed. A-427 human lung cancer cells were infected in vitro and mixed with uninfected cells at different ratios. A-427 tumors in nude mice (n = 23) were injected with 1 x 10(6) to 5 x 10(8) plaque-forming units (pfu) of Ad-hSSTr2-TK. The expressed hSSTr2 and TK proteins were imaged owing to internally bound, or trapped, technetium 99m ((99m)Tc)-labeled hSSTr2-binding peptide (P2045) and radioiodinated 2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl-5-iodouracil (FIAU), respectively. Iodine 125 ((125)I)-labeled FIAU was used in vitro and iodine 131 ((131)I)-labeled FIAU, in vivo. The (99m)Tc-labeled P2045 and (125)I- or (131)I-labeled FIAU were imaged simultaneously with different window settings with an Anger gamma camera. Treatment effects were tested with analysis of variance.

RESULTS

Infected cells in culture trapped (125)I-labeled FIAU and (99m)Tc-labeled P2045; uptake correlated with the percentage of Ad-hSSTr2-TK-positive cells. For 100% of infected cells, 24% +/- 0.4 (mean +/- SD) of the added (99m)Tc-labeled P2045 was trapped, which is significantly lower (P <.05) than the 40% +/- 2 of (125)I-labeled FIAU that was trapped. For the highest Ad-hSSTr2-TK tumor dose (5 x 10(8) pfu), the uptake of (99m)Tc-labeled P2045 was 11.1% +/- 2.9 of injected dose per gram of tumor (thereafter, dose per gram), significantly higher (P <.05) than the uptake of (131)I-labeled FIAU at 1.6% +/- 0.4 dose per gram. (99m)Tc-labeled P2045 imaging consistently depicted hSSTr2 gene transfer in tumors at all adenovirus doses. Tumor uptake of (99m)Tc-labeled P2045 positively correlated with Ad-hSSTr2-TK dose; (131)I-labeled FIAU tumor uptake did not correlate with vector dose.

CONCLUSION

The hSSTr2 and TK proteins were simultaneously imaged following dual gene transfer with an adenovirus vector.

摘要

目的

用无创γ相机成像比较两种评估基因向癌细胞和异种移植肿瘤转移的系统。

材料与方法

构建编码人2型生长抑素受体（hSSTr2）和单纯疱疹病毒胸苷激酶（TK）酶的无复制能力腺病毒（Ad-hSSTr2-TK）。体外感染A-427人肺癌细胞，并以不同比例与未感染细胞混合。给23只裸鼠体内的A-427肿瘤注射1×10⁶至5×10⁸个腺病毒Ad-hSSTr2-TK的空斑形成单位（pfu）。由于内部结合或捕获的锝99m（⁹⁹ᵐTc）标记的hSSTr2结合肽（P2045）和放射性碘化的2'-脱氧-2'-氟-β-D-阿拉伯呋喃糖基-5-碘尿嘧啶（FIAU），分别使表达的hSSTr2和TK蛋白显影。体外使用碘125（¹²⁵I）标记的FIAU，体内使用碘131（¹³¹I）标记的FIAU。用Angerγ相机以不同的窗设置同时对⁹⁹ᵐTc标记的P2045和¹²⁵I或¹³¹I标记的FIAU进行成像。用方差分析测试治疗效果。

结果

培养中的感染细胞捕获¹²⁵I标记的FIAU和⁹⁹ᵐTc标记的P2045；摄取与Ad-hSSTr2-TK阳性细胞的百分比相关。对于100%的感染细胞，添加的⁹⁹ᵐTc标记的P2045有24%±0.4（均值±标准差）被捕获，这显著低于（P<.05）被捕获的¹²⁵I标记的FIAU的40%±2。对于最高的Ad-hSSTr2-TK肿瘤剂量（5×10⁸ pfu），⁹⁹ᵐTc标记的P2045的摄取量为每克肿瘤注射剂量的11.1%±2.9（此后为每克剂量），显著高于（P<.05）¹³¹I标记的FIAU每克剂量1.6%±0.4的摄取量。在所有腺病毒剂量下，⁹⁹ᵐTc标记的P2045成像始终能描绘出肿瘤中hSSTr2基因的转移情况。⁹⁹ᵐTc标记的P2045在肿瘤中的摄取与Ad-hSSTr2-TK剂量呈正相关；¹³¹I标记的FIAU在肿瘤中的摄取与载体剂量无关。