Identification of tyrosine kinases overexpressed in head and neck cancer
Identification of tyrosine kinases overexpressed in head and neck cancer
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DOI：
10.1001/archotol.130.3.311
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发表时间：
2004-03-01
影响因子：
--
通讯作者：
Sun, ZJ
中科院分区：
文献类型：
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作者：
Lin, HS;Berry, GJ;Sun, ZJ
目的：鉴定可能参与头颈部鳞状细胞癌（HNSCC）发生发展的蛋白酪氨酸激酶（PTKs）。设计：将7例HNSCC标本的信使RNA逆转录为互补DNA，利用退化的PTK引物进行聚合酶链反应（PCR），实现PTK互补DNA的选择性扩增。然后克隆这7个HNSCC标本的PTK PCR产物并随机选择进行测序。在这些随机选择的克隆中多次出现的ptk被选为可能在HNSCC中过表达的候选ptk。针对这些候选PTKs的抗体随后被用于其他8个未用于原始候选PTKs选择的HNSCC标本的免疫组织化学研究。结果：基于退化引物逆转录酶PCR技术，在7例HNSCC样本中发现了3个已知PTKs （EphAl， Brk和Ron）和2个新PTKs （KIAA0728和KIAA0279）高表达。免疫组织化学研究显示，在其他8例未用于之前逆转录- pcr反应的HNSCC标本中，EphA1、Brk和Ron在12.5%、37.5%和75%的标本中过表达。结论：在本研究中，我们利用逆转录- pcr技术鉴定出5个在HNSCC中过表达的PTKs，并在8个档案HNSCC标本中证实了3个已知PTKs的过表达。我们的发现提示通过EphA1、Brk和Ron介导的信号通路可能参与了HNSCC的发生和发展。
Objective: To identify protein-tyrosine kinases (PTKs) that may be involved in the development and progression of head and neck squamous cell carcinoma (HNSCC).Design: Messenger RNA from 7 HNSCC specimens was reverse transcribed to complementary DNA, and selective amplification of PTK complementary DNA was achieved using polymerase chain reaction (PCR) with degenerate PTK primers. The resulting PTK PCR products from these 7 HNSCC specimens were then cloned and randomly selected for sequencing. The PTKs that were represented multiple times in these randomly selected clones were selected as candidate PTKs that may be overexpressed in HNSCC. Antibodies against these candidate PTKs were then used for immunohistochemical studies on 8 other HNSCC specimens not used in the original selection of the candidate PTKs.Results: Three known (EphAl, Brk, and Ron) and 2 novel (KIAA0728 and KIAA0279) PTKs were found to be highly expressed in the 7 HNSCC samples studied, based on the technique of reverse transcriptase PCR with degenerate primers. Immunohistochemical studies with antibodies against the 3 known PTKs in 8 other HNSCC specimens not used in the previous reverse transcriptase-PCR reaction demonstrated overexpression of EphA1, Brk, and Ron in 12.5%, 37.5%, and 75% of these specimens.Conclusions: In this study, we identified 5 PTKs that were overexpressed in HNSCC using a reverse transcriptase-PCR technique and confirmed the overexpression of 3 known PTKs in some of the 8 archival HNSCC specimens studied. Our finding suggests that the signaling pathways mediated through EphA1, Brk, and Ron may be involved in the development and progression of HNSCC.