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Non-small-cell lung cancer metastasis and miRNA marker of pre-judging risk of non-small-cell lung cancer metastasis

A non-small cell lung cancer, marker technology, applied in the field of miRNA markers

Inactive Publication Date: 2016-01-27
AFFILIATED HOSPITAL OF GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The lack of effective early diagnosis and treatment methods for non-small cell lung cancer metastasis and drug resistance is the bottleneck in the current prevention and treatment

Method used

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  • Non-small-cell lung cancer metastasis and miRNA marker of pre-judging risk of non-small-cell lung cancer metastasis
  • Non-small-cell lung cancer metastasis and miRNA marker of pre-judging risk of non-small-cell lung cancer metastasis
  • Non-small-cell lung cancer metastasis and miRNA marker of pre-judging risk of non-small-cell lung cancer metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Cell Transfection

[0054] The A549 cells were grouped into inhibition negative control group (anti-NC), miR-340 inhibition group (anti-miR-340) and miR-340 group, and the inhibition negative control group and miR-340 inhibition group were transfected with anti -NC and anti-miR-340 were transfected by conventional methods using transfection reagents, and cells in each group were collected 48 hours after transfection for subsequent experiments.

Embodiment 2Q

[0055] Embodiment 2QPCR experiment

[0056] Add an appropriate amount of Trizol to each group of cells obtained in Example 1 (600 μL per well in a 6-well plate, 1 mL Trizol in a 10 cm petri dish), repeatedly blow and beat to fully lyse the cells, then lyse on ice or at 4°C for 10 min, and collect the lysate in an RNase-free EP tubes can be stored at -20°C for short-term storage.

[0057] QPCR detection of miR-340 expression, the results are as follows figure 1 As shown, compared with the anti-NC inhibition negative control group, the miR-340 level in the miR-340 inhibition group (anti-miR-340) was significantly decreased, indicating that anti-miR-340 can effectively inhibit the expression of miR-340.

Embodiment 3

[0058] Example 3 DAPK protein expression level analysis

[0059] The cell lysate preserved in Example 2 was used to extract the protein for testing according to the requirements of Western blotting, and the expression of DAPK and GAPDH was detected by Western blotting. The result is as figure 2 As shown, the results showed that miR-340 targeted to interfere with the expression of DAPK, and anti-miR-340 could inhibit the function of miR-340 and increase the level of DAPK.

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Abstract

The invention discloses application of miR-340 as a marker of detecting non-small-cell lung cancer metastasis, or as a marker of pre-judging the risk of non-small-cell lung cancer metastasis, or as a drug resistance marker for non-small-cell lung cancer. It is discovered that the miR-340 can target at downregulation of the expression level of death-associated protein kinase, the death-associated protein kinase being closely related to the metastasis and drug resistance of non-small-cell lung cancer. The increase in expression of the miR-340 indicates the increase in the chances of non-small-cell lung cancer metastasis, causing drug resistance and bad prognosis. On this basis, the moiR-340 can also serve as a target for inhibiting / preventing non-small-cell lung cancer adhesion or attack or inhibiting / preventing non-small-cell lung cancer drug resistance and can also be further popularized and applied to the prevention, treatment and prognosis of other tumor cells. The miR-340 is a new diagnostic tool provided for the clinical molecular-level diagnosis of non-small-cell lung cancer metastasis and drug resistance, and a new drug target is provided for the genetic therapy of non-small-cell lung cancer metastasis.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the metastasis of non-small cell lung cancer and a miRNA marker for predicting the risk of metastasis. Background technique [0002] Non-small cell lung cancer (NSCLC) is a complex disease that seriously threatens human life and health. It can occur in almost all parts, tissues, and organs of the human body. The treatment of non-small cell lung cancer is a multidisciplinary comprehensive treatment, in which chemotherapy is one of the important means of treatment, but the drug resistance of non-small cell lung cancer and metastasis of non-small cell lung cancer are the reasons for the low overall survival rate of non-small cell lung cancer patients. reason. Lack of effective early diagnosis and treatment for non-small cell lung cancer metastasis and drug resistance is the bottleneck in the current prevention and treatment. [0003] Death-associated protein kinase is a calc...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/68A61K48/00A61K31/7105A61P35/00
Inventor 伍俊张海涛熊禹真
Owner AFFILIATED HOSPITAL OF GUANGDONG MEDICAL UNIV
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