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Application of TMIGD2 gene in diagnosing and treating rectal adenocarcinoma

A rectal adenocarcinoma and gene technology, applied in gene therapy, medical preparations containing active ingredients, microbial measurement/inspection, etc., can solve problems such as tumor metastasis

Pending Publication Date: 2016-04-06
成都望路医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, despite great progress in early diagnosis and comprehensive treatment, tumor metastasis and recurrence are still the primary reasons affecting prognosis

Method used

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  • Application of TMIGD2 gene in diagnosing and treating rectal adenocarcinoma
  • Application of TMIGD2 gene in diagnosing and treating rectal adenocarcinoma
  • Application of TMIGD2 gene in diagnosing and treating 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.

[0064] 2. Preparation of RNA samples (operated using QIAGEN tissue RNA extraction kit)

[0065] 1) Tissue extraction

[0066] In a clean area with less RNase interference, use a mortar containing an appropriate amount of liquid nitrogen to weigh the isolated intervertebral disc nucleus pulposus tissue sample, grind it to powder with a pestle, and then transfer the sample to a RNase-free 2ml centrifuge tube middle. Add 300 μl of lysate, place in a homogenizer, grind thoroughly for 5 minutes, centrifuge at 12,000 g at 4°C for 10 minutes, and transfer the supernatant to a new 1.5ml centrifuge tube. Add 600 μl of RNase-free water, mix w...

Embodiment 2

[0084] Example 2 QPCR sequencing verification of differential expression of TMIGD2 gene

[0085] 1. According to the detection results of high-throughput sequencing, the TMIGD2 gene was selected for large-sample QPCR verification. According to the sample collection method in Example 1, 80 cases of non-metastatic rectal adenocarcinoma tissue and 80 cases of metastatic rectal adenocarcinoma tissue were selected.

[0086] 2. The RNA extraction steps are the same as in Example 1.

[0087] 3. Reverse transcription: use the reverse transcription kit of TAKARA company to operate. Specific steps are as follows:

[0088] (1) Take 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.

[0089] (2) Construct a 25 μl reaction system, including 5 μl of 5× reverse transcription buffer, 5 μl of dNTP (2.5 mM), 40 U / μl of RNasin, 200 U / μl of M-MLV, and make up to the expected volume with nuclease-...

Embodiment 3

[0110] Embodiment 3 suppresses TMIGD2 gene expression

[0111] 1. Cell culture: human rectal adenocarcinoma cell line SW480, in DMEM medium containing 10% calf serum and 1% P / S at 37°C, 5% CO 2 , in an incubator with a relative humidity of 90%. The medium was changed once every 2-3 days, and 0.25% trypsin was used for routine digestion and passage.

[0112] 2. siRNA design

[0113] siRNA sequences against TMIGD2:

[0114] siRNA1-TMIGD2:

[0115] The sense strand is 5'-UUUGCUUUUUCUUCACAAGCC-3' (SEQ ID NO.7);

[0116] The antisense strand is 5'-CUUGUGAAGAAAAAGCAAAAA-3' (SEQ ID NO.8),

[0117] siRNA2-TMIGD2:

[0118] The sense strand is 5'-UUUUUUGUGUGUACCUAUGGG-3' (SEQ ID NO.9);

[0119] The antisense strand is 5'-CAUAGGUACACACAAAAAAAGG-3' (SEQ ID NO.10),

[0120] siRNA3-TMIGD2:

[0121] The sense strand is 5'-UUUUUGUUUUUUUUGAGACGG-3' (SEQ ID NO.11);

[0122] The antisense strand is 5'-GUCUCAAAAAAAAACAAAAAGC-3'(SEQ ID NO.12)

[0123] Negative control siRNA sequence (si...

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Abstract

The invention discloses a molecular marker related to rectal adenocarcinoma, and particularly relates to application of a TMIGD2 gene in diagnosing and treating rectal adenocarcinoma. The invention also discloses application of the TMIGD2 gene in preparing products for predicting the risk of rectal adenocarcinoma metastasis and diagnosing rectal adenocarcinoma metastasis, and application of the TMIGD2 gene in preparing drugs for treating rectal adenocarcinoma metastasis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of TMIGD2 gene in the diagnosis and treatment of rectal adenocarcinoma. Background technique [0002] Rectal adenocarcinoma is one of the most common malignant tumors in human beings. Its incidence rate in the world is the second among females and the third among males. With the changes in living habits and dietary structure, the incidence of rectal adenocarcinoma in my country is also increasing year by year, which seriously threatens the life and health of Chinese residents. The occurrence and development of colorectal cancer is a multi-stage and multi-step development process involving multiple genes and factors. Its formation has a clear stage, and adenoma is a stage of carcinogenesis. However, from a biological point of view, the malignant proliferation of tumors is mainly manifested in two aspects: one is the indestructibility of tumor cells, that is, cell apo...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61K48/00A61K31/713A61P35/00
CPCC12Q1/6886A61K31/713A61K45/00C12Q2600/118C12Q2600/158G01N33/57446G01N33/68
Inventor 杨承刚林慧芳
Owner 成都望路医药技术有限公司
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