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Gene markers for lung cancer

A technology for labeling and lung cancer, applied in the determination/testing of microorganisms, anti-animal/human immunoglobulin, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve high medical expenses, adverse Outcomes, lack of lung cancer antibodies, etc.

Inactive Publication Date: 2004-01-28
RESONAC CORP +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, lung cancer markers such as cytokeratin-19 and CEA are used to diagnose NSCLC by RT-PCR (reverse transcriptase polymerase chain reaction), but they lack specificity and lead to a large number of false positives and false negative
[0004] However, RT-PCR of micrometastases is particularly advantageous for the detection of lung cancer because of the large patient population, high incidence of metastases via blood, poor prognosis, and high cost of advanced cancer therapy. Medical expenses
In addition, there are no specific antibodies for lung cancer

Method used

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  • Gene markers for lung cancer
  • Gene markers for lung cancer
  • Gene markers for lung cancer

Examples

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example 1

[0038] s-AFLP differential display

[0039] s-AFLP is based on selective PCR amplification of restriction fragments from cDNA libraries. Two sets of selective primers were used. The first set is selective primer A shown in Figure 1, which includes three parts; a core sequence, an enzyme-specific sequence and two selective nucleotides at the 3' end. It can provide 42 = 16 kinds of primers due to the change of selective primers. The second set is the optional primer B shown in Figure 1, which also includes three parts: an anchor sequence, a polyT sequence and two optional nucleotides labeled with fluorophores at the 3' end. It can provide 3 x 4 = 12 primers because there are three selection sequences after polyT (A, G, and C). Therefore, all 3' terminal cDNA restriction fragments including polyA fragments were included in the 192 (16 x 12) types generated by this selective PCR method. Each signal that appeared on the gel display of the amplified fragments represented a non-ove...

example 2

[0055] candidate gene

[0056] Candidate markers are listed in Table 1. The first candidate marker found was identified as the syndecan 1 gene (nucleotide sequence PubMed deposit number BC008765, protein sequence deposit number AAH08765) (SEQ ID NOS: 18 and 19). Figure 6 provides the sequence of the fragment produced by s-AFLP (SEQ ID NO: 22). A blast search of the sequence yielded a paired fragment of exons 2-6 of the syndecan 1 gene (GenBank accession number Z48199). Syndecan 1 is a cell surface transmembrane heparan sulfate proteoglycan from the proteoglycan family that binds the extracellular matrix and growth factors. Loss of regulation of this gene has been identified in several cancers.

[0057]Another candidate marker was identified as collagen 1α2 (nucleotide sequence PubMed accession number J00114, protein sequence accession number AAA51996) (SEQ ID NO: 20 and 21). Figure 7 provides the sequences of the old fragment (SEQ ID NO: 24) and the new fragment (SEQ ID NO...

example 3

[0062] Expression of said markers in normal lung cells and lung cell lines

[0063] To determine whether the markers are expressed in normal human lung cells, RT-PCR was performed on normal human lung RNA using small air vent epithelial cells from human lung, small vessel epithelial cells from lung, bronchial epithelial cells and normal human lung adult cells. Equivalent RNA from fibroblasts.

[0064] Preparation of total RNA from normal lung or lung cancer cell lines

[0065] Total RNA was isolated from cells by acid-guanidine thiocyanate / phenol / chloroform extraction using the TRI REAGENT(R) method (Molecular Research Center, Inc., Cincinnati, OH, USA). Exhaustive purification of RNA is not necessary, and in fact, some DNA contamination is acceptable if primers are designed to span introns.

[0066] RT-PCR

[0067] All RNA samples were treated with DNaseI prior to reverse transcription to ensure no genomic DNA contamination. 1 unit of DNaseI (Life Technologies) was used p...

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Abstract

A method for the diagnosis and identification of new or residual lung cancer is disclosed which uses newly identified markers for lung cancer including syndecan 1, collagen 1 alpha 2, and two novel proteins, 7013 and 7018. The method involves identification of the lung cancer markers is blood from a patient. It is envisioned that at least one marker may be used or any mixture of the four. The method may also include the identification of cytokeratin-19.

Description

field of invention [0001] The present invention relates to methods for diagnosing and determining residual lung cancer. The invention also relates to the use of the newly identified cell markers in the diagnosis of lung cancer. The markers included syndecan 1, collagen 1α2, and two novel proteins 7013 and 7018. Background of the invention [0002] Lung cancer is one of the most common cancers in industrialized countries and has a very high mortality rate. At present, early diagnosis and effective treatment cannot be carried out. Chest x-rays are commonly used for lung cancer screening, however, this method is not suitable for detecting early-stage cancers. CT scans can also be used and can detect cancer at an earlier stage, however, this method takes time and carries the risk of exposure. [0003] Cancer cells or micrometastases are often detected in the blood of patients with melanoma, thyroid cancer, and prostate cancer. The existing reverse-transcription-based polyme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574C07H21/04C07K16/30C12N15/09C12Q1/68
CPCG01N33/57423C07H21/04C12Q1/686C12Q1/6809C12Q2600/158G01N33/57484C12Q1/6886C07K16/30C12Q2539/113C12Q2537/162C12Q2525/191C12Q2525/15
Inventor M·米特苏哈施H·卡姆巴拉H·马特苏纳加M·卡瓦穆拉
Owner RESONAC CORP
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