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Use of tmem236 gene in the preparation of neoadjuvant radiotherapy resistance markers for diagnosing rectal cancer

A rectal cancer and multipurpose technology, applied in the field of biology, can solve problems such as few researches, achieve high technical value, wide application potential, and efficient and convenient experimental methods

Active Publication Date: 2022-08-02
广州达安临床检验中心有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

TMEM family genes are generally considered to be tumor suppressors, and many genes in the family are closely related to tumor chemotherapy resistance. However, TMEM236 gene and its relationship with tumor radiotherapy resistance are rarely studied.

Method used

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  • Use of tmem236 gene in the preparation of neoadjuvant radiotherapy resistance markers for diagnosing rectal cancer
  • Use of tmem236 gene in the preparation of neoadjuvant radiotherapy resistance markers for diagnosing rectal cancer
  • Use of tmem236 gene in the preparation of neoadjuvant radiotherapy resistance markers for diagnosing rectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Tumor regression grade (TRG) in patients with locally advanced rectal cancer neoadjuvant radiotherapy

[0041] A total of 12 patients with locally advanced rectal cancer who had received neoadjuvant radiotherapy were collected. A total of 12 cases of tumor tissue and normal tissue samples were matched before and after radiotherapy. The patients were divided into radiosensitive group (TRG=0+1) and radiotherapy resistant group (TRG= 2+3), with normal tissue as the control. Among them, 6 patients were radiosensitive and 6 were radioresistant. Table 1 below presents the raw data of 12 patients with locally advanced rectal cancer neoadjuvant radiotherapy.

[0042] The specific methods are as follows: (Grading is carried out with reference to the AJCC version of TRG grading)

[0043] AJCC version of tumor regression grade (Tumor Regression Grade, TRG)

[0044]

[0045] Table 1 Tumor regression grades in patients with neoadjuvant radiotherapy for rectal cancer (T...

Embodiment 2

[0047] Example 2 Whole exome sequencing of paired tissue samples before and after radiotherapy

[0048] According to the 12 patients with locally advanced rectal cancer in Example 1, the tumor tissue was taken for colonoscopy biopsy before neoadjuvant radiotherapy, and the paired normal tissue was used as a control for whole-exome sequencing. The specific experimental process is as follows: (eg figure 1 , 2 shown)

[0049] Genomic DNA was extracted from the tissue, cut into short fragments by ultrasonic waves, and then connected to the library building adapter for the first amplification. Two rounds of amplification, purified and sequenced on a sequencer.

[0050] Human Whole Exome Sequencing Experimental Process

[0051] 1. Materials and methods

[0052] 1.1 Main instruments

[0053]

[0054] 1.2 Main reagent consumables

[0055]

[0056] 1.3 Enrolled patients and specimen collection

[0057] A total of 12 patients with locally advanced rectal cancer in the Depar...

Embodiment 3

[0208] Example 3 Quality control analysis of whole exome sequencing raw data

[0209] 1. Preprocessing of sequencing raw data

[0210] First, the original image data file obtained by sequencing was used for base calling using the Base Calling software IllunimaCasava 1.8 and converted to obtain the original base sequence (Raw Data), and the result was exported as a FASTQ file. To remove low-quality reads and adapter sequence data, we preprocessed the raw FASTQ sequences to obtain clean reads and ensure the accuracy of subsequent analysis work.

[0211] The data preprocessing and filtering operations are as follows:

[0212] 1) Use Trimmomatic tools (v 0.36) software to process the raw sequencing data with the following options: TRAILING: 20, MINLEN: 235 and CROP: 235 to remove tailing sequences with a phred quality score lower than 20 and obtain sequencing of uniform length fragments data for subsequent analysis.

[0213] 2) Use FastQC software to check the quality of the se...

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Abstract

The present invention provides the use of TMEM236 gene in preparing a marker for diagnosing locally advanced rectal cancer neoadjuvant radiotherapy primary resistance. The TMEM236 gene mutation site is chr10:17796115. If it is detected that the TMEM236 gene is at the site The point mutation of chr10:17796115, TMEM236:NM_001098844:exon4:c.G667A, shows that the patient is resistant to radiotherapy. The present invention also provides the use of the kit for detecting TMEM236 gene mutation in the preparation of a neoadjuvant radiotherapy resistance diagnostic composition for diagnosing locally advanced rectal cancer. The germline mutation mutant gene TMEM236 screened by the invention can be used for predicting the primary resistance of patients with locally advanced rectal cancer to neoadjuvant radiotherapy.

Description

technical field [0001] The invention belongs to the field of biology and relates to a gene diagnostic marker, in particular to the use of TMEM236 gene in preparing a neoadjuvant radiotherapy resistance marker for diagnosing locally advanced rectal cancer. Background technique [0002] Rectal cancer is one of the most common malignant tumors of the digestive tract. The median age of onset is about 45 years old. Because of its deep location in the pelvis, complex anatomical relationship, not easy to complete surgery, and high postoperative local recurrence rate. In recent years, the incidence of young and middle-aged people has been on the rise, which has seriously endangered people's lives and property safety. At present, surgery is still the main treatment method for early rectal cancer. However, for locally advanced rectal cancer, preoperative neoadjuvant radiotherapy combined with surgery is the main treatment strategy. This method can improve the rate of radical resection...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
Inventor 陈琳傅传刚杨旭东吴军生安俊峰曹湛波周主青朱哲
Owner 广州达安临床检验中心有限公司
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