| Xiaoli Dai,Qing Zhang,Huili Zhang,Hongtao Xu. Identifying peptides that specifically bind to MDA-MB-468 breast cancer cells. Oncol Transl Med, 2019, 5: 119-122. |
| 乳腺癌MDA-MB-468细胞特异性结合肽的筛选、鉴定 |
| Identifying peptides that specifically bind to MDA-MB-468 breast cancer cells |
| Received:May 07, 2019 Revised:July 19, 2019 |
| DOI：10.1007/s10330-019-0356-6 |
| 中文关键词: 噬菌体；乳腺癌；特异性结合肽 |
| 英文关键词: phage; breast cancer; specific binding peptide |
| 基金项目: |
| Author Name | Affiliation | E-mail | | Xiaoli Dai* | Clinical Medical College, Jiangsu Medical Vocational College | houxiangcai254@163.com | | Qing Zhang | Clinical Medical College, Jiangsu Medical Vocational College | | | Huili Zhang | Clinical Medical College, Jiangsu Medical Vocational College | | | Hongtao Xu | Clinical Medical College, Jiangsu Medical Vocational College | |
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| 中文摘要: |
| 目的：利用噬菌体展示技术筛选与人乳腺癌MDA-MB-468细胞特异性结合的小分子多肽。方法：以MDA-MB-468细胞为靶细胞，人正常乳腺细胞SC1180细胞为吸附细胞，利用噬菌体展示随机七肽库与MDA-MB-468细胞进行减性体外筛选，提取高亲和力噬菌体DNA，测定氨基酸序列，进行同源性分析。结果：（1）多肽库原库库容为2×1013pfu/ml，进行4次噬菌体滴度测定，其库容量均值为1.8×1013pfu/ml。（2）减性筛选结果发现，每轮噬菌体文库量均保持在1.8×1012，每一轮都比前次有更明显的富集效果，到第4轮时，与第3轮差别不大，中止筛选。噬菌体在乳腺癌细胞的富集程度提高了约51倍。（3）PCR检测结果显示，在全部产物中，约39个产物（78.0%）长度为300bp，11个产物（22.0%）长度为258bp。挑选30个阳性噬菌体进行DNA提取并测序，翻译出氨基酸序列为LMTRXSK。同源性分析结果显示，该序列与中已知基因和蛋白无同源性。结论：利用噬菌体展示技术体外筛选获得了能与人乳腺癌MDA-MB-468细胞特异性结合的小分子多肽LMTRXSK，有可能成为乳腺癌诊断及靶向治疗潜在靶点。 |
| 英文摘要: |
| Objective To use phage display technique to screen for small polypeptides that specifically bind to MDAMB-468 cells.
Methods A random heptapeptide phage display library was used for in vitro screening against target
MDA-MB-468 cells. SC1180 cells were used for subtractive selection. High-affinity phage DNA was
extracted, and peptides were sequenced.
Results (1) The original library capacity of the polypeptide library was 2 × 1013 pfu/mL, and phage
titer was determined over 4 rounds. The average library capacity was 1.8 × 1013 pfu/mL. (2) Subtractive
screening showed that the phage library volume of each round was 1.8 × 1012 pfu/mL, and that there was an
enrichment effect in each subsequent round. Screening was stopped after the fourth round. (3) PCR results
showed that the size of 39 products (78.0%) and 11 products (22%), were 300 bp and 258 bp, respectively.
Thirty positive phages were selected for DNA extraction and sequencing, and the corresponding amino acid
sequence was LMTRXSK. The sequence had no homology with known genes or proteins.
Conclusion Using the phage display technique, we identified that the short polypeptide, LMTRXSK,
specifically binds MDA-MB-468 human breast cancer cells. |
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