<sup>99m</sup> Tc-HYNIC-PTP的制备及其胰腺癌裸小鼠移植瘤模型SPECT显像研究

龚佳丽; 赵凌舟; 赵晋华

doi:10.19732/j.cnki.2096-6210.2021.02.002

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^99m Tc-HYNIC-PTP的制备及其胰腺癌裸小鼠移植瘤模型SPECT显像研究

论著 | 更新时间：2025-12-15

- ^99m Tc-HYNIC-PTP的制备及其胰腺癌裸小鼠移植瘤模型SPECT显像研究
- Preparation of ^99m Tc-HYNIC-PTP and its SPECT imaging in nude mice bearing pancreatic cancer xenografts
- 肿瘤影像学 2021年30卷第2期页码：71-77
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金（81801727）
- DOI：10.19732/j.cnki.2096-6210.2021.02.002
  中图分类号：
- 网络出版：2021-04-28，
  
  纸质出版：2021-04-28
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龚佳丽,赵凌舟,赵晋华. ^99m Tc-HYNIC-PTP的制备及其胰腺癌裸小鼠移植瘤模型SPECT显像研究[J]. 肿瘤影像学, 2021, 30(2): 71-77 https://doi.

org/10.19732/j.cnki.2096-6210.2021.02.002
龚佳丽,赵凌舟,赵晋华. ^99m Tc-HYNIC-PTP的制备及其胰腺癌裸小鼠移植瘤模型SPECT显像研究[J]. 肿瘤影像学, 2021, 30(2): 71-77 https://doi. DOI： 10.19732/j.cnki.2096-6210.2021.02.002.

org/10.19732/j.cnki.2096-6210.2021.02.002 DOI：

摘要

目的：

制备

99m

Tc-肼基烟酰胺（hydrazino-nicotinamide，HYNIC）-网蛋白靶向肽（plectin targeted peptide，PTP），探讨其用于胰腺癌单光子发射计算机断层成像（single photon emission computed tomography，SPECT）的可行性。

方法：

PTP多肽N端赖氨酸残基游离的氨基分别与HYNIC和异硫氰酸荧光素（fluorescein isothiocyanate，FITC）反应，制备HYNIC-PTP和FITC-PTP。在三（羟甲基）甲基甘氨酸和乙二胺二乙酸的磷酸盐缓冲生理盐水（phosphate-buffered saline，PBS；pH=6.0）体系中，以SnCl

为还原剂，100 C反应10 min标记

99m

Tc，得到

99m

Tc-HYNIC-PTP标志物。利用快速薄层色谱（instant thin layer chromatography，ITLC）分析放射化学纯度，以及在室温PBS和37 C胎牛血清中的稳定性。细胞增殖-毒性试验［细胞计数试剂盒-8（cell counting kit-8，CCK-8）］评价检测HYNIC-PTP多肽体外对BxPC-3细胞增殖的影响；蛋白质印迹法（Western blot）比较胰腺癌细胞BxPC-3、PANC-1与小鼠胰岛素瘤胰岛细胞（-TC-6细胞）内网蛋白（plectin）的表达量；流式细胞术分析FITC-PTP与BxPC-3细胞的体外结合靶向性。建立皮下胰腺癌裸小鼠移植瘤模型，经尾静脉注射

99m

Tc-HYNIC-PTP后，在不同时间点（0.5、1.0、2.0、4.0和6.0 h）进行SPECT，分析肿瘤摄取情况。

结果：

ITLC分析表明，

99m

Tc-HYNIC-PTP放射化学纯度大于99%，且体外稳定性良好；Western blot证实，plectin在BxPC-3与PANC-1细胞中都高度表达，远高于阴性对照-TC-6细胞（

＜0.001）；流式细胞术结果显示，FITC-PTP在阳性对照PANC-1细胞和BxPC-3细胞中的荧光信号强度无明显差异，表明FITC-PTP对BxPC-3细胞具有良好的靶向性。SPECT显像表明，

99m

Tc-HYNIC-PTP在BxPC-3胰腺癌裸小鼠移植瘤模型中

具有良好的肿瘤摄取，注射0.5 h后肿瘤部位即有显影，1.0 h后明显增强，2.0 h显影清晰，4.0 h有所下降，6.0 h后肿瘤部位仍有清楚显影。

结论：

本研究成功制备了

99m

Tc-HYNIC-PTP，方法简便、放射化学纯度高、稳定性好，在胰腺癌裸小鼠移植瘤模型中具有明显的肿瘤浓聚，有潜力成为一种胰腺癌分子影像探针。

Abstract

Objective:

To prepare

99m

Tc-labeled hydrazino-nicotinamide (HYNIC) plectin targeted peptide (PTP) and investigate its feasibility as a single photon emission computed tomography (SPECT) imaging agent for pancreatic cancer.

Methods:

The N-terminal of PTP was modified with HYNIC and fluorescein isothiocyanate (FITC) to form HYNIC-PTP and FITC-PTP

which are respectively used for radiolabeling of

99m

Tc and evaluation of targeting ability by flow cytometry.

99m

Tc-HYNIC-PTP was prepared in a phosphate-buffered saline (PBS

pH=6.0) with the presence of co-ligands tricine and ethylenediamine diacetate at 100 C for 10 min

SnCl 2 as reducing agent. The radiochemical purities of

99m

Tc-HYNIC-PTP in PBS (at room temperature) and fetal bovine serum (at 37℃) at different time points was tested using instant thin layer chromatography (ITLC) to evaluate the stability

in vitro

. The potential cytotoxicity of HYNIC-PTP was analyzed using cell counting kit-8 (CCK-8) assays. Western blot was used to analyzethe expression level of plectin protein in both BxPC-3 and PANC-1 pancreatic cancer cells

and -TC-6 cells were set as negative control. The targeting of FITC-PTP to BxPC-3 cells was determined by flow cytometry

and PANC-1 cells were used as a positive control. SPECT images were acquired to observe the tumor uptake at 0.5

1.0

2.0

4.0 and 6.0 h after intravenous injection of

99m

Tc- HYNIC-PTP into nude mice bearing pancreatic cancer xenografts.

Results:

99m

Tc-HYNIC-PTP has high radiochemical purity (＞99%) and good stability in vitro. The plectin protein expression in both BxPC-3 and PANC-1 cells were much higher than that of -TC-6 ce

lls. The flow cytometry results showed that the target binding rate of FITC-PTP was also high in BxPC-3 cells when compared to PANC-1 cells. As for tumor accumulation

the SPECT signal intensity in tumor region was observed at 0.5 h post- injection and further enhanced at 1.0 h post-injection. The strongest tumor signal intensities appeared at 2.0 h post-injection

followed by a gradual decrease in tumor accumulation at 4.0 and 6.0 h post-injection.

Conclusion:

99m

Tc-HYNIC-PTP was successfully obtained using a facile preparation method with high radiochemical purity and stability in vitro. SPECT imaging showed that

99m

Tc- HYNIC-PTP could obviously accumulate in the tumor region

suggesting great potential to be developed as a molecular imaging probe for pancreatic cancer.

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99m Tc-HYNIC-PTP的制备及其胰腺癌裸小鼠移植瘤模型SPECT显像研究

Preparation of 99m Tc-HYNIC-PTP and its SPECT imaging in nude mice bearing pancreatic cancer xenografts

DOI：10.19732/j.cnki.2096-6210.2021.02.002

摘要

Abstract

关键词

Keywords

references

^99m Tc-HYNIC-PTP的制备及其胰腺癌裸小鼠移植瘤模型SPECT显像研究

Preparation of ^99m Tc-HYNIC-PTP and its SPECT imaging in nude mice bearing pancreatic cancer xenografts