Preparation of 68Ga-NOTA-CD44 peptide and assessment of its targeting ability towards CD44 + triple-negative breast cancer stem cells
10.3760/cma.j.cn321828-20240429-00148
- VernacularTitle:68Ga－NOTA－CD44多肽的制备及其靶向CD44 ＋三阴性乳腺癌干细胞的研究
- Author:
Rui YANG
1
;
Ruiying ZHU
;
Chen SU
;
Kai CHENG
;
Jie ZHOU
;
Zhen JIA
;
Mengting DA
;
Jiuda ZHAO
;
Daozhen CHEN
Author Information
1. 江南大学附属妇产医院、南京医科大学附属无锡妇幼保健院优生优育遗传研究所，无锡 214002
- Publication Type:Journal Article
- Keywords:
Breast neoplasms;
Isotope labeling;
Gallium radioisotopes;
Antigens, CD44;
Stem cells;
Tumor cells, cultured
- From:
Chinese Journal of Nuclear Medicine and Molecular Imaging
2025;45(5):294-299
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To conduct enrichment and biological behavior studies on CD44 + CD24 - triple-negative breast cancer (TNBC) stem-like cells, and to construct 68Ga-labeled CD44 peptide ( 68Ga-1, 4, 7-triazacyclononane-1, 4, 7-triacetic acid (NOTA)-CD44p) and evaluate its targeting ability towards the surface marker CD44 of TNBC stem-like cells. Methods:Suspension sphere culture method was utilized to enrich and cultivate CD44 + CD24 - cell subpopulations from TNBC cell line MDA-MB-231 and non-TNBC cell line MCF-7. Flow cytometry was used to detect the expression of stem cell markers of different groups, cell scratch assay was performed to assess the migration ability of CD44 + CD24 - cell subpopulations, and Transwell invasion assay was performed to evaluate the invasion ability of CD44 + CD24 - cell subpopulations. 68Ga-NOTA-CD44p was prepared, followed by purification and identification with high-performance liquid chromatography (HPLC). The targeting ability of 68Ga-NOTA-CD44p towards CD44 + TNBC cells was evaluated through cellular uptake and blocking experiments. Data were analyzed by independent-sample t test, one-way analysis of variance and the least significant difference t test. Results:Suspension sphere culture successfully enriched CD44 + CD24 - TNBC stem-like cell spheres. Compared to the non-TNBC cell line MCF-7, TNBC cell line MDA-MB-231 exhibited better sphere-forming ability (18.50±3.73 vs 31.83±4.92; t=5.29, P<0.001) and a higher proportion of CD44 + CD24 - cell subset ((24.97±8.12)% vs (90.93±4.46)%; F=170.10, t=14.93, both P<0.001). The wound healing rate ((71.00±11.00)% vs (28.33±4.16)%; F=42.91, t=8.02, both P<0.001) and invasion rate ((60.60±16.87)% vs (24.16±8.15)%; F=11.83, t=4.40, both P<0.01) of CD44 + CD24 - MDA-MB-231 group cells were significantly increased compared to the CD44 + CD24 - MCF-7 group. MDA-MB-231 cells showed strong uptake ability of 68Ga-NOTA-CD44p, which decreased after CD44p blocking. Conclusions:Compared to CD44 + CD24 - MCF-7 cells, CD44 + CD24 - MDA-MB-231 cells exhibit higher malignant biological behavior. 68Ga-NOTA-CD44p targets the surface marker CD44 of TNBC stem-like cells, laying the research foundation for targeted therapy against TNBC with tumor stem cells as targets.
