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131Ⅰ-B2-S22-AFA对HER-2高表达乳腺癌细胞的杀伤作用
封面论文The killing effect of 131Ⅰ-labeled B2-S22-AFA on breast cancer cells with high HER-2 expression
- 核技术 2017年40卷第12期 文章编号：120301
- 作者机构：
南昌大学第一附属医院 核医学科 南昌 330006
- 作者简介：
[ "李亚梅, 女, 1974年出生, 2008年于南昌大学获硕士学位, 研究领域为妇产科影像学", "LI Yamei, female, born in 1974, graduated from Nanchang University with a master's degree in 2008, focusing on obstetrics and gynecology imaging" ]
关晏星, E-mail:yanxingguan2000@aliyun.com GUAN Yanxing, E-mail:yanxingguan2000@aliyun.com
- 基金信息：江西省赣鄱英才555工程项目及江西省自然科学基金项目(2007GZY1419)
DOI：10.11889/j.0253-3219.2017.hjs.40.120301
中图分类号： R817.8收稿日期：2017-03-21，
修回日期：2017-10-19，
纸质出版日期：2017-12-10
- 稿件说明：
移动端阅览
李亚梅, 关晏星, 陈学忠, 等. 131Ⅰ-B2-S22-AFA对HER-2高表达乳腺癌细胞的杀伤作用[J]. 核技术, 2017,40(12):120301
Yamei LI, Yanxing GUAN, Xuezhong CHEN, et al. The killing effect of 131Ⅰ-labeled B2-S22-AFA on breast cancer cells with high HER-2 expression[J]. Nuclear techniques, 2017, 40(12): 120301
李亚梅, 关晏星, 陈学忠, 等. 131Ⅰ-B2-S22-AFA对HER-2高表达乳腺癌细胞的杀伤作用[J]. 核技术, 2017,40(12):120301 DOI： 10.11889/j.0253-3219.2017.hjs.40.120301.
Yamei LI, Yanxing GUAN, Xuezhong CHEN, et al. The killing effect of 131Ⅰ-labeled B2-S22-AFA on breast cancer cells with high HER-2 expression[J]. Nuclear techniques, 2017, 40(12): 120301 DOI： 10.11889/j.0253-3219.2017.hjs.40.120301.
摘要
人类表皮生长因子受体2（Human Epidermal Growth Factor Receptor 2，HER-2）是乳腺癌最重要的致病基因。本文探讨
131
Ⅰ标记的HER-2特异结合小分子模拟肽B2-S22-AFA对乳腺癌细胞的特异性杀伤作用。采用溴代琥珀酰亚胺法（N-Bromide Succinimide，NBS）进行
131
Ⅰ标记B2-S22-AFA，测定标记率、放化纯度、稳定性及免疫活性。用Western Blot法和免疫组化法测定人乳腺癌SKBR3及MDA-MB-231Her-2细胞HER-2表达，观察不同浓度表皮生长因子（Epidermal Growth Factor，EGF）条件下B2-S22-AFA对细胞生长的抑制率。设B2-S22-AFA组（B2-S22-AFA终浓度1 μg·mL
-1
）、
131
Ⅰ-B2-S22-AFA组（
131
Ⅰ-B2-S22-AFA终浓度144 kBq·μg
-1
·mL）、
131
Ⅰ组（
131
Ⅰ终浓度144 kBq·mL
-1
）、阴性对照组及试剂空白组，每组EGF终浓度均5.0 pg·mL
-1
，采用四唑盐比色法（Methyl Thiazolyl Tetrazolium，MTT）测定SKBR3和MDA-MB-231细胞增殖活性，观察
131
Ⅰ-B2-S22-AFA、B2-S22-AFA及
131
Ⅰ作用不同时间对两种细胞生长的抑制率。结果显示，HER-2在SKBR3细胞呈强阳性表达，在MDA-MB-231细胞呈弱（或低）表达。
131
Ⅰ-B2-S22-AFA标记率、放化纯度及比活度分别为64.8%-79.6%、95.0%-97.6%、151.7 MBq·mg
-1
，在37℃血清中放置4 h、12 h、24 h、48 h、72 h的放化纯度分别95.4%、93.5%、91.4%、88.2%、77.4%，
131
Ⅰ-B2-S22-AFA与SKBR3细胞及MDA-MB-231细胞的最大结合率分别（66.47±3.24）%和（3.89±0.81）%（3次）。当EGF存在时，B2-S22-AFA可明显抑制SKBR3细胞生长；无EGF时，B2-S22-AFA无此作用。
131
Ⅰ-B2-S22-AFA对SKBR3细胞的杀伤作用较B2-S22-AFA和
131
Ⅰ明显增强（均
p
<
0.05），B2-S22-AFA对SKBR3细胞杀伤作用显著高于
131
Ⅰ（
p
<
0.05），
131
Ⅰ-B2-S22-AFA和B2-S22-AFA对MDA-MB-231细胞均无杀伤作用。综上所述，
131
Ⅰ-B2-S22-AFA可显著加强、加速B2-S22-AFA对HER-2高表达乳腺癌细胞的特异性杀伤作用。
Abstract
Background
2
B2-S22-AFA is a kind of small molecule mimetic polypeptide that can conjugate to epidermal growth factor receptor 2 (HER-2) specifically and has obvious depressant effect on breast cancer cells with overexpressing HER-2.
Purpose
2
This study aims to investigate the specific killing effect of
131
Ⅰ-labeled B2-S22-AFA on breast cancer cell with overexpressing HER-2.
Methods
2
131
Ⅰ-B2-S22-AFA was prepared by using N-bromosuccinimide method to measure the labeling efficiency
radiochemical purity
stability
and immunocompetence. The expression levels of HER-2 in SKBR3 and MDA-MB-231 cells were detected by Immunohistochemistry and Western Blot. The inhibitory rate of B2-S22-AFA on cell growth was observed with epidermal growth factors (EGF) at different concentrations. Five groups i.e
B2-S22-AFA group (B2-S22-AFA final concentration:1 μg·mL
-1
)
131
Ⅰ-B2-S22-AFA group (
131
Ⅰ-B2-S22-AFA final concentration:144 kBq/1 μg·mL
-1
)
131
Ⅰ group (
131
Ⅰ final concentration:144 kBq·mL
-1
)
negative control group and reagent blank group
were selected for contrast tests with the final concentration of EGF in each group at 5.0 pg·mL
-1
. The proliferation and activity of cells were measured by Methyl thiazolyl tetrazolium assay. The inhibitory rates were compared among
131
Ⅰ-B2-S22-AFA group
B2-S22-AFA group and
131
Ⅰ group in the growth of the two kinds of cells at different time.
Results
2
1) HER-2 receptors were strongly expressed in SKBR3 cells while weakly (low) or no expressed in MDA-MB-231 cells. The radiochemical purity
labeling rate and specific activity of
131
Ⅰ-B2-S22-AFA were 95.0%-97.6%
64.8%-79.6% and 151.7 MBq·mg
-1
respectively. Radiochemical purity of
131
Ⅰ-B2-S22-AFA was 95.4%
93.5%
91.4%
88.2% and 77.4% when it was placed at 37℃ in serum for 4 h
12 h
24 h
48 h and 72 h
respectively. The maximum binding rates of SKBR3 and MDA-MB-231 cells were (66.47±3.24)% and (3.89±0.81)% (tested for 3 times) respectively. 2) The inhibitory effect of B2-S22-AFA on the growth of SKBR3 cells only in the presence of EGF. 3) The killing effect of
131
Ⅰ-B2-S22-AFA on SKBR3 cells was significantly stronger than that of B2-S22-AFA and
131
Ⅰ (
p
<
0.05)
and the inhibitory effect of B2-S22-AFA on the growth of SKBR3 cells was significantly higher than that of
131
Ⅰ (
p
<
0.05).
131
Ⅰ-B2-S22-AFA and B2-S22-AFA had no obvious killing effect on MDA-MB-231 cells.
Conclusion
2
131
Ⅰ-B2-S22-AFA can significantly enhance and accelerate the specific killing effect of B2-S22-AFA on breast cancer cells with high HER-2 expression.
关键词
Keywords
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