Mobile website
Prompt Report
Inhibitory effect of 131I-labeled anti-CD133 ScFv on CD133＋ human hepatocellular carcinoma cells
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective:To study the inhibitory effect of the anti-CD133 single chain variable fragment (ScFv) labeled with 131I on CD133＋cancer stem cells (CSCs) sorted form human hepatocellular liver carcinoma HepG2 cells in vitro. Methods: CD133+ and CD133- CSCs were isolated from HepG2 cells through magnetic-activated cell sorting (MACS). CD133 expression in both sorted and unsorted cells was analyzed by flow cytometry (FCM). The property of CD133＋ CSCs was validated by sphere-forming assay and colony formation assay in vitro and tumor formation assay in nude BALB/c mice in vivo. The monoclonal antibody CD133 was labeled with 131I using the chloramines T method and the labeling rate, specific activity and radioactivity were evaluated. CD133+ CSCs were treated with 131I, CD133 ScFv, 131I-CD133 ScFv, and 131I＋CD133 ScFv. At 12, 24 and 48 hours after treatment, cell proliferation and cell cycle progression were assessed by MTT assay and FCM respectively. Results: CD133 was detected in (97.71±1.13)% of the sorted CD133＋ HepG2 cells but in only (1.52±0.78)% of unsorted HepG2 cells (P＝0.0001). As compared with CD133- HepG2 cells, CD133＋ HepG2 cells showed a higher tumor sphere formation ability (\[45.03±1.35\]% vs \[7.4±054\]%, P<0.001). The 131I labeling rate of CD133 ScFv was 88.92%, and the radiochemical-purity was 98.63%. A maximal CD133+ cell growth inhibition of (89.58±0.74)% was observed (P<0.05) when 131I was used at 3.7 MBq/100 μl and CD133 ScFv was used at 1 μg/100 μl, significantly higher than other doses (P<0.05). The proportion of G0/G1 phase arrest in cells treated with 131I-CD133 ScFv was significantly reduced as compared with treatments (P<005). Conclusion: Radioisotope 131I labeled CD133 ScFv may effectively inhibit growth of CD133-positive human hepatocellular carcinoma cells in vitro.
Keywords:
Project Supported:
Project supported by the National Natural Science Foundation of China (No.30370422，No.81171365)
Clc Number:
