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Markers for diagnosis and treatment of rectal adenocarcinoma

A technology for rectal adenocarcinoma and expression products, which can be used in gene therapy, biological testing, drug combination and other directions, and can solve the problems of lost operation opportunity, difficult to find, and hidden clinical symptoms of rectal adenocarcinoma.

Active Publication Date: 2018-09-07
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The early clinical symptoms of rectal adenocarcinoma are hidden and difficult to find. Most of the patients are in the middle and late stage when they see a doctor, and the best time for surgery is lost

Method used

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  • Markers for diagnosis and treatment of rectal adenocarcinoma
  • Markers for diagnosis and treatment of rectal adenocarcinoma
  • Markers for diagnosis and treatment of rectal adenocarcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Screening for gene markers associated with rectal adenocarcinoma

[0062] 1. Collection of samples

[0063] 8 cases of rectal adenocarcinoma non-metastatic tissue and rectal adenocarcinoma metastatic tissue samples were collected, and all the above samples were obtained with the consent of the organizational ethics committee. All surgical resection specimens were cut into small pieces with sterilized ophthalmic scissors, put into 1.5ml sterile EP tubes soaked in 0.1% DEPC water overnight and sterilized, and the remaining tissues were stored in cryopreservation tubes and quickly put into Store in a liquid nitrogen tank at -80°C.

[0064] 2. Preparation of RNA samples

[0065] 1) Sample pretreatment: Take the sample from the -80°C liquid nitrogen tank and put it into a homogenizer soaked in 0.1% DEPC water overnight and sterilized, and grind it into powder; during the grinding process, the sample must always be in the In a liquid nitrogen environment, the grind...

Embodiment 2

[0085] Example 2 QPCR sequencing to verify the differential expression of SLC26A9 gene

[0086] 1. According to the detection results of high-throughput sequencing, the SLC26A9 gene was selected for large-sample QPCR verification. According to the sample collection method in Example 1, 70 cases of non-metastatic rectal adenocarcinoma tissue and 70 cases of metastatic rectal adenocarcinoma tissue were selected.

[0087] 2. The RNA extraction steps are the same as in Example 1.

[0088] 3. Reverse transcription:

[0089] 1) Reaction system:

[0090] Reagent

volume

MgCl 2

2μl

10×RT Buffer

1μl

RNase-free water

3.75μl

dNTP mix

1μl

RNase inhibitor

0.25μl

AMV reverse transcriptase

0.5μl

Random 9mers

0.5μl

experimental sample RNA

1μl

[0091] 2) Reverse transcription reaction conditions

[0092] According to the reverse transcription reaction conditions in RNA PCR Kit (AMV) ...

Embodiment 3

[0114] Example 3 Inhibition of SLC26A9 Gene Expression

[0115] 1. Cell culture (human rectal adenocarcinoma cell line SW480):

[0116] 1) Cell recovery

[0117] a. Take out the cell cryopreservation tube from the liquid nitrogen tank, immediately put the cell cryopreservation tube into warm water at 37°C, and shake it gently to melt it as soon as possible.

[0118] b. Take out the cryopreservation tube from the warm water at 37°C, suck out the cell suspension with a pipette, add 5ml of L-15 culture medium containing 10% bovine serum to the centrifuge tube, mix and centrifuge at 800rpm for 5min, remove the supernatant, Wash again with culture medium.

[0119] c. Add 5ml of L-15 culture solution containing 10% fetal bovine serum, inoculate in a 75ml culture bottle, and place at 37°C, 5% CO 2 cultured in a constant temperature incubator.

[0120] 2) Cell replacement

[0121] a. When the culture solution starts to turn yellow, take out the culture bottle from the incubator a...

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Abstract

The invention discloses a marker SLC26A9 for diagnosing and treating rectal adenocarcinoma. The experiment proves that the SLC26A9 is highly expressed in metastasis rectal adenocarcinoma tissues. The expression level of the SLC26A9 can be detected to predict the risk of rectal adenocarcinoma metastasis, diagnose the metastasis of the rectal adenocarcinoma and judge whether rectal adenocarcinoma relapses. The invention also discloses application of the SLC26A9 gene in preparing drugs for inhibiting / preventing metastasis of rectal adenocarcinoma.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of SLC26A9 gene in the diagnosis and treatment of rectal adenocarcinoma. Background technique [0002] Rectal adenocarcinoma is one of the common malignant tumors of the digestive system. In recent years, with the improvement of people's living standards, changes in diet structure, population aging and the development of rectal adenocarcinoma census, the incidence of rectal adenocarcinoma in my country has been increasing year by year, seriously threatening people's health. And because most of rectal adenocarcinoma has no special symptoms or no symptoms at all in the early stage, the diagnosis of rectal adenocarcinoma is often delayed. When tumor symptoms such as anemia, weight loss, blood in the stool, and abdominal pain appear, the cancer is often in the middle and late stages. Rectal adenocarcinoma has high morbidity and mortality. According to statistics, there...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/68G01N33/574A61K45/00A61K48/00A61K31/713A61P35/00
CPCA61K31/713A61K45/00C12Q1/6886C12Q2600/118C12Q2600/158G01N33/57446G01N33/68
Inventor 杨承刚任静
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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