Phage display library selection of a hypoxia-binding scFv antibody for liver cancer metabolic marker discovery.
Phage display library selection of a hypoxia-binding scFv antibody for liver cancer metabolic marker discovery.
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用于肝癌代谢标志物发现的缺氧结合 scFv 抗体的噬菌体展示文库选择
DOI：
10.18632/oncotarget.9460
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发表时间：
2016-06-21
期刊：
影响因子：
--
通讯作者：
Zhang S
中科院分区：
文献类型：
--
作者：
Liu J;Zhang Q;Chen H;Gao Z;Li Y;Sun Z;Xiang R;Zhang S
在肝癌和转移中经常观察到的缺氧影响肿瘤进展和对治疗的抵抗。尽管缺氧相关的生物标志物在其他癌症中也有用，但没有一种被认为是肝癌缺氧的替代物。在这项研究中，我们产生了七个独特的人单链抗体（scFv）特异性缺氧肝癌细胞（Abs），使用常氧耗尽与低氧选择噬菌体文库淘选技术。通过开发基于scFv免疫沉淀的质谱法，与其中一种Ab（H103）结合的抗原被鉴定为丙酮酸激酶（PKM 2）的M2剪接同种型，PKM 2是癌细胞中有氧糖酵解的关键调节剂。低氧诱导肝癌细胞PKM2表达增加。免疫组化（IHC）染色显示PKM2在中分化和高分化肝细胞癌（HCC）组织中高表达，呈低血管染色模式。PKM 2在肝内胆管癌（ICC）组织中的高表达也定位于周围坏死区。PKM2表达的HCC或ICC肿瘤的百分比在肿瘤坏死较多、微血管密度低和晚期时显著较高。此外，H103 scFv Ab被有效地内化到缺氧的肝癌细胞中，并且可能具有靶向药物递送的潜力。结论：我们的研究首次开发了针对肝癌细胞的缺氧特异性scFv Ab H103，并揭示PKM 2是HCC和ICC组织中缺氧的有希望的生物标志物。这些允许进一步探索这种有价值的Ab和PKM2抗原用于肝癌中的缺氧靶向。
Hypoxia, which is frequently observed in liver cancer and metastasis, influences tumor progression and resistance to therapy. Although hypoxia-associated biomarkers are of use in other cancers, none is recognized as a surrogate for hypoxia in liver cancer. In this study, we generated seven unique human single-chain Fv (scFv) antibodies (Abs) specific to hypoxic liver cancer cells, using normoxia-depleted vs hypoxia-selected phage library panning technology. By developing the scFv immunoprecipitation-based mass spectrometry method, the antigen that bound with one of the Abs (H103) was identified as the M2 splice isoform of pyruvate kinase (PKM2), an enzyme that is a key regulator of aerobic glycolysis in cancer cells. Increased expression of PKM2 was induced by hypoxia in liver cancer cell lines. Immunohistochemical (IHC) staining showed that PKM2 was highly expressed in moderately and well differentiated hepatocellular carcinoma (HCC) tissues with a hypovascular staining pattern. High expression of PKM2 was also localized in the perinecrotic area of intrahepatic cholangiocarcinoma (ICC) tissues. The percentage of the HCC or ICC tumor expressing PKM2 was significantly higher with more tumor necrosis, low microvessel density, and advanced stage. Moreover, the H103 scFv Ab was efficiently internalized into hypoxic liver cancer cells and could have potential for targeted drug delivery. Conclusion: our study, for the first time, developed hypoxia-specific scFv Ab H103 to liver cancer cells, and revealed that PKM2 is a promising biomarker for hypoxia in HCC and ICC tissues. These allow further exploration of this valuable Ab and PKM2 antigen for hypoxia targeting in liver cancer.
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影响因子：
2.5
作者：
Kim Y;Lin Q;Glazer PM;Yun Z
通讯作者：
Yun Z
影响因子：
4.7
作者：
Hacker, HJ;Steinberg, P;Bannasch, P
通讯作者：
Bannasch, P
影响因子：
7.3
作者：
Israelsen WJ;Vander Heiden MG
通讯作者：
Vander Heiden MG
影响因子：
3.4
作者：
Hammond, E. M.;Asselin, M. -C.;Williams, K. J.
通讯作者：
Williams, K. J.
DOI：
10.1073/pnas.92.9.3938
发表时间：
1995-04-25
影响因子：
11.1
作者：
DEKRUIF, J;TERSTAPPEN, L;LOGTENBERG, T
通讯作者：
LOGTENBERG, T