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Applications of FAM176A gene in diagnosis and treatment for rectum adenocarcinoma

A technology of FAM176A, 1.FAM176A, which can be used in medical preparations containing active ingredients, biochemical equipment and methods, and determination/inspection of microorganisms, which can solve problems such as recurrence

Active Publication Date: 2016-04-27
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In clinical treatment, surgery is the main treatment, supplemented by comprehensive treatment such as chemotherapy, radiotherapy or targeted therapy, but about 30%-50% of patients undergo surgery, radiotherapy, chemotherapy and other traditional methods within 2-3 years. May recur
[0004] At present, there is no marker that can be used as an independent tissue diagnosis, and it becomes very necessary to find a new specific marker for the diagnosis, prognosis and treatment of rectal adenocarcinoma

Method used

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  • Applications of FAM176A gene in diagnosis and treatment for rectum adenocarcinoma
  • Applications of FAM176A gene in diagnosis and treatment for rectum adenocarcinoma
  • Applications of FAM176A gene in diagnosis and treatment for rectum adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Screening for gene markers associated with rectal adenocarcinoma

[0061] 1. Sample collection

[0062] Paracancerous tissue and rectal adenocarcinoma tissue samples were collected from 8 cases of rectal adenocarcinoma. The above samples are surgical resection specimens of patients with rectal adenocarcinoma, and all the above samples were obtained with the consent of the organizational ethics committee.

[0063] 2. RNA sample preparation (using miRNAkit for operation)

[0064] The tissues obtained above were shredded, put into liquid nitrogen and ground into powder, and RNA was extracted and isolated according to the instructions in the kit. details as follows:

[0065] (1) Isolation of RNA:

[0066] ① Add in tissue homogenate or cells Reagent II 1ml;

[0067] ②Leave at room temperature for 3min, add 0.2ml chloroform and shake vigorously for 15s;

[0068] ③Put it on ice for 10 minutes;

[0069] ④ Centrifuge at 12000g for 15min at 4°C;

[0070] ⑤Tr...

Embodiment 2

[0085] Example 2 QPCR sequencing to verify the differential expression of the FAM176A gene

[0086] 1. Large-sample QPCR verification of differential expression of FAM176A gene. According to the sample collection method in Example 1, 70 cases of rectal adenocarcinoma paracancerous tissues and 70 cases of rectal adenocarcinoma tissues were selected.

[0087] 2. The specific operation steps of QPCR are as follows:

[0088] (1) RNA extraction

[0089] The RNA extraction procedure was as described in Example 1.

[0090] (2) Reverse transcription: the reverse transcription kit from TAKARA Company was used for operation. Specific steps are as follows:

[0091] ①Take 1 μg of total RNA for reverse transcription, add 2 μl of Oligo(dT), and mix well. Immediately after 5 min in water bath at 70°C, ice bath for 2 min.

[0092] ② Construct a 25 μl reaction system, including 5 μl of 5× reverse transcription buffer, 5 μl of dNTP (2.5 mM), RNasin 40 U / μl, M-MLV 200 U / μl, and add nucleas...

Embodiment 3

[0113] Overexpression of embodiment 3FAM176A gene

[0114] 1. Cell culture (human rectal adenocarcinoma HRC-99)

[0115] (1) Culture cells in an incubator at 37°C, 5% CO2, and 90% relative humidity with RPMI1640 medium containing 10% calf serum and 1% P / S.

[0116] (2) When the cells grow to 70%-90%, they are routinely digested and passaged with 0.25% EDTA-containing trypsin.

[0117] (3) Subculture every 2-3 days.

[0118] 2. Overexpression of FAM176A gene

[0119] (1) Construction of FAM176A gene expression vector

[0120] Amplification primers were designed according to the coding sequence of the FAM176A gene (as shown in SEQ ID NO.1), and the primer sequences were as follows:

[0121] Forward primer: 5'-CCGAAGCTTGCCACCATGAGGCTGCCCCTCAGCC-3' (SEQ ID NO.7)

[0122] Reverse primer: 5'-CGGGCGGCCGCATAGTAGCGATTCAGGCT-3' (SEQ ID NO.8)

[0123] The coding sequence of the full-length FAM176A gene was amplified from the cDNA library of adult fetal brain (clontech company, prod...

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Abstract

The invention discloses applications of FAM176A gene in diagnosis and treatment for rectum adenocarcinoma. The experiment proves that the FAM176A gene is in low expression in the tissue of the patient suffering from rectum adenocarcinoma; and the adding of the expression of FAM176A can inhibit the proliferation of rectum adenocarcinoma cells and promote the apoptosis of the rectum adenocarcinoma cells. According to the invention, a diagnosis means for the rectum adenocarcinoma is discovered, and meanwhile, a novel target spot is provided for the gene therapy of the rectum adenocarcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of FAM176A gene in the diagnosis and treatment of rectal adenocarcinoma. Background technique [0002] Rectal adenocarcinoma is a common malignant tumor of the digestive tract in humans with various histological types. In some developed countries, the mortality rate of rectal cancer has jumped to the second place among all malignant tumors, while in developing countries, the incidence rate is also rising due to the influence of certain factors. The rapid rise has made it one of the fastest-rising malignant tumors. Rectal adenocarcinoma has a high degree of malignancy and is prone to infiltration and metastasis, which affects the quality of life and endangers life. With the changes in people's lifestyle and diet structure, the incidence of rectal adenocarcinoma is increasing year by year, and the age of onset is also tending to be younger. The proportion of young and midd...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61K31/7088A61P35/00
CPCA61K31/7088A61K45/00C12Q1/6886C12Q2600/158G01N33/57446G01N33/68
Inventor 董东王欣月杨承刚
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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