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Colorectal cancer prognostics

A colorectal cancer, prognosis technology, applied in antineoplastic drugs, biochemical devices and methods, drug combinations, etc., can solve problems such as difficult to detect benefits, controversial, low recurrence rate, etc.

Inactive Publication Date: 2013-07-17
VERIDEX LCC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the relatively low recurrence rate, the benefit of postoperative chemotherapy in Duke's B patients is difficult to detect and remains controversial.
However, due to certain flaws in the grading of Duke's B stage, approximately 20-30% of such patients behave more like Duke's stage C and relapse within 5 years

Method used

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  • Colorectal cancer prognostics
  • Colorectal cancer prognostics
  • Colorectal cancer prognostics

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Sample Processing and LCM

[0051] Fresh-frozen tissue samples were collected from patients with surgically resected colorectal cancer tumors. The samples used were from 63 patients classified as Duck's B stage according to standard diagnostics and pathology. The clinical outcome of the patients is known. Thirty-six patients were recurrence-free for more than 3 years, while 27 patients experienced tumor recurrence within 3 years.

[0052] Tissues were quickly frozen with liquid nitrogen within 20-30 minutes of harvesting and then stored at -80°C. To enable laser capture, samples were sectioned (6 μm), one mounted on a glass slide and the other mounted on a film (P.A.L.M.) that had been fixed on a glass slide (Micro Slides Colorfrost, VWR Scientific, Media , PA). Slide-mounted sections were fixed in cold acetone and stained with Mayer's hematoxylin (Sigma, St. Louis, MO). Pathologists analyze samples for diagnosis and grading. A clinical grade assess...

Embodiment 2

[0055] Example 2 RNA extraction and amplification

[0056] Total RNA was extracted from LCM-captured samples using a Zymo-Spin Column (Zymo Research, Orange, CA 92867). Approximately 2 ng of total RNA was resuspended in 10 μl of water and subjected to 2 rounds of amplification using T7 RNA polymerase to obtain approximately 50 μg of amplified RNA.

Embodiment 3

[0057] Embodiment 3 DNA microarray hybridization and quantification

[0058] Samples were assayed by a DNA microarray panel consisting of approximately 23,000 human DNA clones using commercially available human U133a from Affymetrix. The total RNA prepared and obtained according to the aforementioned method was used for the chip, and analyzed using Agilent BioAnalyzer referring to the product manual. All 63 samples met quality control criteria and their data were used for marker selection.

[0059] The chip intensity data was analyzed by the software MAS VERSION 5.0 of Fffymetrix Company. Automated analysis identified two genes that could differentiate relapsed from non-relapsed patients.

[0060] The chip intensity data obtained above were used as the input value of commercial automatic clustering software such as PARTEK 5.1. A high recurrence rate cluster of 20 patients (13 relapsed, 7 survivors) was identified using this automated clustering algorithm. From the initial ...

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Abstract

Prognosis of colorectal cancer The present invention provides a method for judging the prognosis of colorectal cancer by analyzing the expression of a group of genes. These include gene expression patterns displayed on various media such as microarrays, and kits containing gene expression patterns.

Description

technical background [0001] This application is a continuation-in-part of US Application No. 10 / 651,237, filed August 28, 2003. [0002] The present invention relates to the prognosis of colorectal cancer based on gene expression patterns of biological samples. [0003] Colorectal cancer is a heterogeneous disease with complex origins. The probability of recurrence in patients with colorectal cancer after treatment is related to the degree of tumor infiltration in the intestinal wall and the presence or absence of nodular involvement. These factors are also the basis for the current Duke's staging. Duke's stage A disease is limited to the submucosa of the colon or rectum. Duke's stage B tumors invade the muscle layer and can invade the bowel wall of the rectum or colon. Duke's stage C disease includes the presence of regional lymph node metastases and any degree of intestinal wall invasion. [0004] Surgical resection is highly effective in treating early-stage colorectal ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/106C12Q2600/118A61P35/00C12Q1/6837
Inventor Y·王
Owner VERIDEX LCC