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. 2012 May;15(5):287-93.
doi: 10.3779/j.issn.1009-3419.2012.05.07.

[Proteomics-based identification of proapoptotic caspase adapter protein as a novel serum marker of non-small cell lung cancer]

[Article in Chinese]
Heng Zhang  1 , Shengxi Chen, Lingjin Huang
Affiliations

[Proteomics-based identification of proapoptotic caspase adapter protein as a novel serum marker of non-small cell lung cancer]

[Article in Chinese]
Heng Zhang et al. Zhongguo Fei Ai Za Zhi. 2012 May.

Abstract
in English, Chinese

Background and objective: Early diagnosis and treatment of lung cancer contribute to a patient's prolonged survival. Proteomics and tissue culture were used to screen potential serum markers of lung cancer.

Methods: The differential proteomic technique and matrix-assisted laser desorption/ionization-time of flight mass spectrometry were used to identify overexpressed proteins from cultured tumor tissues in a conditioned medium. Paired lung tissues in a conditioned medium were used as the control group. Overexpressed proapoptotic caspase adaptor protein (PACAP) was identified using Western blot and enzyme-linked immunosorbent assay (ELISA).

Results: The expression of nineteen protein spots exhibited a twofold increase compared with that in the control group. Fourteen of which were identified, including the distinctly overexpressed PACAP. Western blot analysis revealed a high level of PACAP in the lung adenocarcinoma in both tissue culture media and patients' serum. Overexpressed PACAP was also detected in the serum of non-small cell lung cancer patients via ELISA compared with that in the serum of small cell lung cancer patients, benign lung tumor patients, and healthy individuals (P<0.05).

Conclusion: PACAP is a potential serum marker of non-small cell lung cancer.

背景与目的: 早诊断早治疗可以提高肺癌的长期生存率，血清标志物可能有助于肺癌的早期诊断。本研究在组织培养的基础上，应用蛋白质组学方法筛选潜在的肺癌血清标志物。

方法: 采用差异蛋白质组学技术对肺腺癌和肺组织培养液中的差异蛋白质进行初步鉴定。选取其中的半胱天冬酶凋亡前衔接蛋白（proapoptotic caspase adapter protein, PACAP）做更进一步的研究，用Western blot验证PACAP在培养液上清中的表达情况并检测它在不同类型血清中的表达情况，用ELISA检测不同类型血清标本中PACAP的表达情况。

结果: 共得到差异蛋白点19个，鉴定出14个。PACAP在双向电泳中表达差异较大，Western blot显示PACAP在肺癌组织培养液及肺癌患者血清中的表达均明显增高。ELISA显示PACAP在肺腺癌和肺鳞癌患者血清中的表达水平高于在小细胞肺癌、肺良性肿瘤和健康人血清中的水平（P < 0.05）。

结论: PACAP是潜在的非小细胞肺癌血清标志物。

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Figures

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1
肺腺癌培养液上清蛋白（A）及配对肺组织培养液上清蛋白（B）双向凝胶电泳考染图谱（图中点1-19是人属蛋白质） Coomassie blue dyed 2D Gel electrophoresis of conditioned medium of lung cancer tissue (A) and paired lung tissue (B) (Spot 1-19 are homo sapiens)
2
2
差异蛋白点6号在二维电泳中的表达情况局部图（A代表腺癌组，B代表肺组织组） The partial detail of the sixth differential expressed protein spot in 2D Gel electrophoresis (A stands for lung carcinoma conditioned medium and B stands for that of lung tissue)
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PACAP的PMF及Mascot搜索结果。A：通过MALDI-TOF质谱仪得到的肽指纹图。B：搜索数据库得到的与PACAP相吻合的5个肽段质谱峰。C：PACAP氨基酸序列，粗体字母代表质谱检测到的肽段序列与PACAP的匹配情况。 Identification of PACAP by peptide mass fingerprint. A: Peptide mass spectrum was obtained by MALDI-TOF mass spectrometry; B: The masses of five typical peptides were matched with PACAP by database searching; C: The sequence of PACAP is represented by single-letter code for amino acids. Sequence coverage by five peptides is indicated with bold capital letters.
4
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PACAP在4组肺腺癌（Xca）和配对肺组织（Xf）培养液上清中的Western blot结果 Highly expressed PACAP in lung adenocarcinoma tissues conditioned medium. The expression levels of PACAP were compared by immunoblotting in conditioned medium between lung adenocarcinoma (Xca) and paired lung tissue (Xf).
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PACAP在6组肺腺癌（Xca）和正常人血清（Xf）标本中的Western blot结果 Highly expressed PACAP in lung adenocarcinoma serum. The expression levels of PACAP were compared by immunoblotting in serum between lung adenocarcinoma (Xca) and health people (Xf).
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