Application of ALDH18A1 in treatment and diagnosis of colorectal cancer

A technology for ALDH18A1 and colorectal cancer, applied in gene therapy, medical preparations containing active ingredients, biochemical equipment and methods, etc., can solve unclear problems

Active Publication Date: 2019-03-01
THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the clinicopathological significance of ALD

Method used

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  • Application of ALDH18A1 in treatment and diagnosis of colorectal cancer
  • Application of ALDH18A1 in treatment and diagnosis of colorectal cancer
  • Application of ALDH18A1 in treatment and diagnosis of colorectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. ALDH18A1 is highly expressed in colorectal cancer and has prognostic value

[0031] (1) ALDH18A1 is highly expressed in colorectal cancer tissues

[0032] Log in to Xena Public Data Hubs (http: / / xena.ucsc.edu / public-hubs / ) to download colorectal cancer RNA sequencing data (TCGA Colon And Rectal Cancer, COADREAD), the chip platform is IlluminaHiSeq 2000RNA Sequencing platform; via R data package (affy 1.42.3) Download the GSE41258 dataset of the GEO database, the chip platform is Affymetrix Human Genome U133A Array, and the expression profile difference analysis uses the R language package (limma 3.20.9).

[0033] Organize the expression data of the GSE41258 dataset, take the median expression level of ALDH18A1 as the critical value, divide into ALDH18A1 high / low expression group, and make phenotype data files. The GSE41258 dataset showed that ALDH18A1 mRNA expression levels were significantly increased in colorectal cancer patients (average expression 8.365±0....

Embodiment 2

[0048] Example 2, the effect of shRNA knockdown on the expression of ALDH18A1 gene on the proliferation activity of colorectal cancer cells (1) cell culture

[0049] Human colorectal cancer cell lines HT29, HCT116 and normal colonic epithelial cells NCM460 were placed in DMEM high-glucose medium containing 10% fetal bovine serum at 37°C and 5% CO 2 , Cultured in a constant temperature incubator with 95% humidity, changed the medium every other day, and passaged in 3 to 5 days.

[0050] (2) Packaging and infection of ALDH18A1 shRNA lentivirus

[0051] ALDH18A1 shRNA (5'-ccggccattatttgaccagatcattctcgagaatgatctggtcaaataatggttttg-3'(SEQ ID NO.1), 5'-aattcaaaaaccattatttgaccagatcattctcgagaatgatctggtcaaataatgg-3'(SEQ ID NO.2)) (Sigma-Aldrich-LV-RNA, USA) was constructed using molecular cloning technology In the lentiviral vector (Yingmaoshengye, Beijing), named ALDH18A1 shRNA. Using Lipofectamine 3000 transfection reagent (Invitrogen, USA), 5 μg ALDH18A1 shRNA, 3.75 μg PSPAX2 and 1...

Embodiment 3

[0052] Example 3. Correlation between ALDH18A1 expression and oncogene MYC

[0053] (1) There is a positive correlation between ALDH18A1 and MYC at the mRNA level

[0054] Studies have found that almost all colorectal cancer patients have varying degrees of uncontrolled expression of the MYC gene, which plays an important role in the malignant proliferation, stemness maintenance, and cancerous transformation of colorectal cancer. Therefore, we next explore whether the regulation of ALDH18A1 on the proliferation ability of colorectal cancer cells is related to the MYC gene.

[0055] Extract the mRNA expression information of ALDH18A1 and MYC in the GSE41258 data set of colon cancer patients, and use Real-Time PCR to detect the mRNA levels of ALDH18A1 and MYC. Meter measures RNA concentration. According to Prime ScriptRT Reagent Kit (Takara) instructions, the above extracted RNA was reverse transcribed into cDNA, and followed by SYBR Premix ExTaq TM II instructions, take 7.5...

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Abstract

The invention relates to an application of ALDH18A1 in treatment and diagnosis of colorectal cancer. The ALDH18A1 is highly expressed in colorectal cancer tissue and related to clinical pathological parameters and prognosis of the colorectal cancer tissue; cell proliferation of the colorectal cancer can be inhibited through down-regulation of expression of the ALDH18A1; the ALDH18A1 is in positivecorrelation with expression of an MYC gene; MYC gene expression can be inhibited through down-regulation of the ALDH18A1; in patients with high expression of the ALDH18A1, c-MYC target genes are significantly enriched; the ALDH18A1 can be used as a prognosis factor for colorectal cancer patients, can inhibit proliferation of colorectal cancer cells by down-regulating expression of the MYC gene and has great significance in tretament and diagnosis of the colorectal cancer.

Description

technical field [0001] The invention belongs to the field of disease diagnosis and treatment, and specifically relates to the application of ALDH18A1 in the treatment and diagnosis of colorectal cancer. Background technique [0002] Colorectal cancer is one of the most common malignant tumors, with more than 1.2 million newly diagnosed patients worldwide every year. According to the American Cancer Society (ACS), there were about 140,000 new cases in 2018, accounting for 8.1% of all cancer cases, and about 50,000 deaths, accounting for 8.3% of cancer deaths. At present, the 5-year overall survival rate of colorectal cancer patients is 64.5%, and about 25% of patients have metastases at the time of diagnosis, while the 5-year overall survival rate of patients with metastases is only 14%. Traditional surgical treatment and adjuvant chemotherapy are still the main treatment methods for patients with colorectal cancer. Therefore, finding new therapeutic targets and strategies ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/573G01N33/574A61K48/00A61K31/7088A61K31/713A61P35/00
CPCA61P35/00G01N33/573G01N33/57407G01N33/57484A61K31/7088A61K31/713C12Q1/6886G01N2333/90203
Inventor 余时沧郭玉峰杨泽宇
Owner THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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