Isolation and identification of CD133-positive liver cancer stem cells and <sup>131</sup>I-CD133mAb biological effects on the cells

侯妍利; 唐敏; 陈兴月; 段丽群; 康强强; 舒锦; 李少林

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Isolation and identification of CD133-positive liver cancer stem cells and ¹³¹I-CD133mAb biological effects on the cells

更新时间：2026-01-09

- Isolation and identification of CD133-positive liver cancer stem cells and ¹³¹I-CD133mAb biological effects on the cells
- Journal of Army Medical University Vol. 36, Issue 3, Pages: 222-227(2014)
- 作者机构：
- 作者简介：
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- DOI：
  CLC： R329-33;R329.28;R735.7
- Published：2014
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侯妍利,唐敏,陈兴月,段丽群,康强强,舒锦,李少林.分离、鉴定CD133+人肝癌干细胞以及131I-CD133mAb对其生物学效应的影响[J].陆军军医大学学报(原第三军医大学学报),2014,36(03):222-227.

　Hou Yanli,Tang Min,Chen Xingyue,Duan Liqun,Kang Qiangqiang,Shu Jin,Li Shaolin.Isolation and identification of CD133-positive liver cancer stem cells and 131I-CD133mAb biological effects on the cells[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(03):222-227.
侯妍利,唐敏,陈兴月,段丽群,康强强,舒锦,李少林.分离、鉴定CD133+人肝癌干细胞以及131I-CD133mAb对其生物学效应的影响[J].陆军军医大学学报(原第三军医大学学报),2014,36(03):222-227. DOI：

　Hou Yanli,Tang Min,Chen Xingyue,Duan Liqun,Kang Qiangqiang,Shu Jin,Li Shaolin.Isolation and identification of CD133-positive liver cancer stem cells and 131I-CD133mAb biological effects on the cells[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(03):222-227. DOI：

摘要

目的分离、鉴定CD133

人肝癌干细胞以及

131

I-CD133mAb对其生物学效应的影响。方法为选择最适的细胞系，流式细胞仪检测Huh-7和HepG2细胞中CD133表达率；磁珠分选Huh-7细胞，流式细胞仪检测分选后CD133表达率；体外成球、克隆形成实验及体内成瘤实验验证干细胞特性；氯胺T法制备

131

I标记CD133单克隆抗体（

131

I-CD133mAb）并鉴定;取分选后的CD133+-Huh-7细胞、Huh-7细胞和HepG2细胞进行实验，将每种细胞分成4组（

131

I-CD133mAb组、

131

I组、CD133mAb组、

131

I+CD133mAb组）；MTT法检测各组对CD133

-Huh-7细胞生长抑制的最适剂量和各组在最适剂量作用后24、48、72 h对3组细胞生长抑制率；流式细胞仪检测

131

I-CD133mAb作用3组细胞72 h时细胞凋亡和细胞周期的变化。结果 Huh-7和HepG2细胞系CD133表达率分别为18.8%和5.2%，故选用Huh-7细胞进行分选；磁珠分选后CD133

-Huh-7细胞CD133的表达率为99.28%，与分选前比较差异有统计学意义（

0.01）。CD133

-Huh-7细胞相对于CD133

-Huh-7细胞具有更强的体外成球能力、克隆形成能力和体内成瘤能力。

131

I-CD133mAb标记率为87.92%，放射化学纯度为97.54%，具有较好的稳定性和免疫活性。当

131

I 4.8 MBq/100 μL、CD133mAb 4.8 μg/100 μL时对CD133+-Huh-7细胞抑制率最高（

0.05），取此放射性浓度进行实验。24、48、72 h时各组对CD133

-Huh-7细胞的抑制作用明显高于Huh-7和HepG2细胞组，

131

I-CD133mAb组对3种细胞生长抑制率明显高于其余各组，抑制率随时间的增加而升高（

0.01）。

131

I-CD133mAb作用CD133

-Huh-7细胞72 h后凋亡率为31.21%，

明显高于其余2组（

0.05）。细胞周期检测结果显示CD133

-Huh-7细胞G

期减少54.77%，明显高于其余2组（

0.05）。结论 CD133

细胞具有肿瘤干细胞特性，

131

I-CD133mAb能特异性结合CD133

-Huh-7细胞，调控细胞周期和诱导细胞凋亡。

Abstract

Objective To study the sorting of CD133

liver cancer cells having the features of cancer stem cells

and to study

131

I-labeled anti-CD133 monoclonal antibody (

131

I-CD133mAb) biological effects on the cells. Methods Flow cytometry (FCM) was used to detect the CD133 expression on Huh-7 cells and HepG2 cells. Magnetic-activated cell sorting (MACS) was used to isolate CD133

and CD133

cells from Huh-7 cells. FCM was used to detect the expression of CD133 after cell isolation. The stem cell properties of sorted CD133

cells were validated by sphere-forming assay and colony formation assay in vitro and BALB/c mice transplantation tumor experiments in vivo.

131

I-CD133mAb was prepared by the chloramines-T method and then identified. CD133

Huh-7 cells

Huh-7 cells and HepG2 cells were separately divided into 4 groups including

131

I-CD133mAb group

131

I group

CD133mAb group and

131

I+CD133mAb group. The optimal inhibitory concentration and the inhibitory effects at 24

and 72 h on cell proliferation in each group were measured by MTT assay. The apoptosis and cell cycle change rates of the three cells 72 h after

131

I-CD133mAb treatment were evaluated by FCM. Results CD133 expression rates on Huh-7 cells

HepG2 cells and CD133

Huh-7 cells were 18.8%

5.2% and 99.28%

respectively. CD133

Huh-7 cells showed a higher tumor sphere formation ability and tumor-genesis capacity compared with CD133

Huh-7 cells. The labeling ratio of

131

I-CD133mAb was 87.92% and the radiochemical-purity was 97.54%. The growth inhibition rate on

CD133

Huh-7 cells was the highest when

131

I was 4.8 MBq/100 μL and CD133mAb was 4.8 μg/100 μL as shown by MTT assay (P0.05). In all groups at 24

72 hours after treatment

the growth inhibitory rate on CD133

Huh-7 cells was significantly higher than that on Huh-7 cells and HepG2 cells and the growth inhibitory rate of

131

I-CD133mAb group was obviously the highest as proven by MTT assay

which showed a time-dependent manner (P0.05). The apoptotic rate of CD133

Huh-7 cells in 72 h after

131

I-CD133mAb treatment was 31.21%

significantly higher than that of other groups (P0.05). The rate of CD133

Huh-7 cells in G

0/G

phase was reduced to 54.77%

significantly greater than that of other groups (P0.05). Conclusion CD133

cells have the characteristics of cancer stem cells.

131

I-CD133mAb can effectively bind with CD133

Huh-7 cells to regulate cell cycle and induce apoptosis.

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Isolation and identification of CD133-positive liver cancer stem cells and 131I-CD133mAb biological effects on the cells

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Isolation and identification of CD133-positive liver cancer stem cells and ¹³¹I-CD133mAb biological effects on the cells