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目的寻找能够与胃癌腹膜高转移细胞GC9811P特异性结合的噬菌体多肽,探索治疗胃癌腹膜转移的新方法。方法运用噬菌体呈现肽技术,先后用胃癌的腹膜高转移细胞系GC9811P和其亲本细胞GC9811对噬菌体12肽库进行消减性的全细胞淘洗,经过3轮筛选,随机挑选40个噬菌体单克隆C1～C40。用ELISA法选取能够与GC9811P特异性结合的单克隆。将选出的单克隆分别注入裸鼠腹腔,采用免疫组化法排除与正常组织亦高结合的阳性单克隆。对筛选出的噬菌体克隆进行DNA序列测定,并推导其外源性氨基酸序列,进行同源性分析。结果经过3轮淘洗,噬菌体克隆得到理想富集。C9、C18、C23、C29、C34和C37可与GC9811P特异性结合,经免疫组化证实,这6个单克隆均不与裸鼠腹腔内正常组织结合。测序结果大致展示了两种外源性多肽,即TLNINRLILPRT和SMSIXSPYIXXX。结论筛选出6个可与GC9811P细胞特异性结合的噬菌体多肽;这两个肽序列能否阻断GC9811P细胞向腹膜转移尚待进一步确定。
Objective To search for a phage polypeptide that can specifically bind to GC9811P in gastric peritoneal metastatic cells and to explore a new method for treating peritoneal metastasis of gastric cancer. Methods Using phage display peptide technology, the phagocyte 12 peptide pool was eluted by whole cell with descending GC9811P and its parent cell GC9811. After three rounds of screening, 40 phage monoclonal C1 ~ C40. A single clone that specifically binds to GC9811P was selected by ELISA. The selected monoclonal were injected into the peritoneal cavity of nude mice, using immunohistochemistry to exclude positive clones also bound to normal tissues. The selected phage clone DNA sequence determination, and deduced its exogenous amino acid sequence, homology analysis. Results After 3 rounds of panning, the phage clones obtained the ideal enrichment. C9, C18, C23, C29, C34 and C37 could specifically bind to GC9811P. Immunohistochemistry confirmed that none of the six monoclonal antibodies bind to the normal tissues of abdominal cavity of nude mice. The sequencing results broadly demonstrate two exogenous polypeptides, TLNINRLILPRT and SMSIXSPYIXXX. Conclusion Six phage polypeptides were screened for specific binding to GC9811P cells. Whether these two peptide sequences can block the metastasis of GC9811P cells to the peritoneum remains to be determined.