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Premier pair and kit for detecting expression state of MT2A in cancer cells, and application of DATS in expression of MT2A

A detection kit and technology for gastric cancer cells, applied in biological testing, DNA/RNA fragments, measuring devices, etc., can solve the problems of lack of early diagnosis and prognosis evaluation of gastric cancer, achieve far-reaching clinical significance and popularization, good stability, The effect of easy operation

Inactive Publication Date: 2014-02-12
BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

The main reasons for this are, on the one hand, bad living habits such as smoking, diet, alcoholism, salted products, and microbial infections such as pylori, and on the other hand, the lack of effective indicators for early diagnosis and prognosis evaluation of gastric cancer

Method used

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  • Premier pair and kit for detecting expression state of MT2A in cancer cells, and application of DATS in expression of MT2A
  • Premier pair and kit for detecting expression state of MT2A in cancer cells, and application of DATS in expression of MT2A
  • Premier pair and kit for detecting expression state of MT2A in cancer cells, and application of DATS in expression of MT2A

Examples

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Embodiment 1

[0045] The IHC staining protein detection kit can contain the following components required for IHC detection: MT2A antibody (for example, E918-0133, Invitrogen Company, the United States), and immunohistochemical related reagents (xylene, alcohol, 5% skimmed milk powder , 3%H 2 o 2 , 1×PBS, secondary antibody of the same species, DAB chromogenic system, hematoxylin nuclear dye and resin mounting medium), and gastric tissue microarray samples. Specifically, the gastric tissue microarray samples used in Example 1 (3) IHC stained protein detection included: 684 gastric cancer samples, 118 intestinal metaplasia samples with precancerous lesions and 171 normal gastric mucosa samples.

[0046] Specifically, in the system for detecting IHC stained proteins using the above-mentioned IHC stained protein detection kit, prepare the primary antibody to MT2A (for example, E918-0133, Invitrogen, USA), at a dilution ratio of 1:50, and dilute it with the primary antibody diluent . The sel...

Embodiment 2

[0063] This example is used to illustrate that DATS can up-regulate the expression of MT2A gene in gastric cancer cells.

[0064] (1) Through the operation steps of semi-quantitative PCR and Western Blot protein detection in Example 1 above, we detected the expression levels of mRNA (318bp) and protein (15kD) of MT2A with different treatment concentrations and treatment times of DATS, half of which The templates used in quantitative PCR and Western Blot protein detection were the cDNA or protein obtained by treating BGC82312h with different concentrations of DATS (0, 25 μM, 50 μM, 100 μM) and the cDNA or protein obtained by treating BGC823 with 25 μM DATS for different times (0h, 0.5h, 2h, 6h, 12h) obtained cDNA or protein.

[0065] The result is as Figure 4 shown. With the increase of DATS treatment concentration, the protein expression of MT2A mRNA was significantly increased. As detected by grayscale scanning, 100 μM DATS treatment increased the expression of MT2A by 7 t...

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Abstract

The invention provides a premier pair and a kit for detecting the expression state of MT2A gene nucleic acid in cancer cells and an application of DATS for increasing the expression of MT2A gene in cancer cells. The premier pair comprises a first premier pair or a second premier pair, the first premier pair comprises a first MT2A premier and a GAPDH premier used as an internal reference premier, and the second premier pair comprises a second MT2A premier, and a gamma-actin premier used as an internal reference premier.

Description

technical field [0001] The invention relates to a pair of primers for detecting the expression state of MT2A gene in gastric cancer cells, a kit for detecting the MT2A gene in gastric cancer cells and the application of DATS in up-regulating the expression of MT2A gene in gastric cancer cells. Background technique [0002] At present, gastric cancer is still one of the main cancers that seriously threaten the health of Chinese people. The incidence of gastric cancer in the world ranks second among all tumors, and the death rate still ranks first. The proportion of gastric cancer patients in China accounts for 44% of the world. The main reasons for this are, on the one hand, unhealthy living habits such as smoking, diet, alcoholism, pickled products, and microbial infections such as pylori, and on the other hand, the lack of effective indicators for early diagnosis and prognosis evaluation of gastric cancer. Although traditional radiotherapy, chemotherapy and surgery can dela...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11G01N33/68
CPCC12Q1/6886C12Q2600/112C12Q2600/118C12Q2600/158G01N33/57446
Inventor 吕有勇潘元明李文梅崔建涛邢蕊
Owner BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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