Application of MGST1 gene in proliferation and apoptosis of lung adenocarcinoma cell

A technique for lung adenocarcinoma cells and lung adenocarcinoma, which is applied in the application field of proliferation and apoptosis, and can solve the problems of low quality of life, poor patient prognosis, and low 5-year survival period.

Inactive Publication Date: 2017-11-24
信雅生物科技(苏州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Lung adenocarcinoma is one of the most common tissue types of lung cancer. The curative effect has not improved significantly in recent ye...

Method used

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  • Application of MGST1 gene in proliferation and apoptosis of lung adenocarcinoma cell
  • Application of MGST1 gene in proliferation and apoptosis of lung adenocarcinoma cell
  • Application of MGST1 gene in proliferation and apoptosis of lung adenocarcinoma cell

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0026] Embodiment 1: MGST1 lentivirus infection

[0027] After the A549 cell line was spread on a 6-well plate for 10 hours, replace it with serum-free RPMI-1640Medium containing 5ug / ul polybree, add the corresponding virus volume (number of cells*MOI / virus titer), and culture for 8 hours. Replace with RPMI-1640Medium containing 10% fetal bovine serum. Fluorescence was detected 72 hours after transfection ( figure 1 ), it can be observed that the lentivirus infection efficiency of the control group and the experimental group is greater than 80%.

specific Embodiment approach 2

[0028] Embodiment 2: Western detection of protein expression

[0029] Collect the cells in a centrifuge tube and centrifuge at 2000rpm for 4min, wash twice with pre-cooled PBS, add an appropriate volume of RIPA (10μl / ml protease inhibitor and phosphatase inhibitor) according to the cell volume, mix by pipetting, and seal the EP tube After the membrane was sealed, put it in a cell fragmentation ultrasonic instrument at 4°C, 500watt, lyse for 20min; centrifuge at 14000g, 20min, to precipitate cell debris, and the supernatant was the whole cell protein, and the concentration was determined by BCA method. The amount of protein loaded per well was 50ug. Electrophoresis conditions 5% stacking gel 80V, 50 minutes; 12% separating gel 120V, 100 minutes. The transfer condition was 100V for 100 minutes. The dilution ratio of MGST1 primary antibody was 1:1000, and the dilution ratio of secondary antibody was 1:8000.

[0030] Test results( figure 2 ) shows: the protein expression of M...

specific Embodiment example 3

[0031] Specific implementation case 3: Inhibiting the expression of MGST1 can significantly inhibit the proliferation of lung adenocarcinoma cell A549

[0032] MTS assay was used to detect the proliferation of human lung adenocarcinoma cell line A549. Take the control group Scramble in the growth index phase and the experimental group shMGST1-85, shMGST1-86 cells, count the cells and inoculate them in a 96-well cell culture plate, with 2000 cells per well, and measure the OD value at a wavelength of 490nm with a full-wavelength scanner. data analysis.

[0033] Experimental results ( image 3 ) shows that compared with the control group Scramble, the proliferation speed of shMGST1-85 and shMGST1-86 cells in the experimental group was significantly slowed down, and as time went by, the difference in the number of viable cells began to be statistically significant, p<0.05, indicating that Inhibiting the expression of MGST1 can significantly inhibit the proliferation of human lu...

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Abstract

The invention relates to application of a human-derived MGST1 gene in promoting proliferation of an A549 lung adenocarcinoma cell line and inhibiting apoptosis of the A549 lung adenocarcinoma cell line. According to the application, by virtue of methods such as knock-down of the A549 lung adenocarcinoma cell line by virtue of a slow virus MGST1, MTS proliferation detection, giemsa plate cloning and dyeing, flow cytometry apoptosis detection and naked mouse subcutaneous tumor formation, the influence of the MGST1 on the proliferation and apoptosis of the A549 lung adenocarcinoma cell line is judged, and results show that proliferation and cell clonality of A549 cells can be inhibited by inhibiting expression of the MGST1, and the early apoptosis of the A549 cells is promoted. A naked mouse experiment shows that the tumor volume of lung adenocarcinoma can be obviously reduced by inhibiting expression of the MGST1 and the tumor growth is inhibited.

Description

technical field [0001] The invention relates to a new application of human MGST1, in particular to the application of human MGST1 gene in the proliferation and apoptosis of lung adenocarcinoma cells. Background technique [0002] Lung cancer is a highly malignant tumor with high morbidity and mortality worldwide, which seriously threatens people's life and health. In my country, the mortality rate due to lung cancer ranks first among all malignant tumors, and it is also the focus and difficulty of current cancer researchers. Lung adenocarcinoma is one of the most common tissue types of lung cancer. The curative effect has not improved significantly in recent years. The 5-year survival period of patients is still very low. The prognosis of patients is still very poor, and the quality of life is very low. Therefore, it is of great significance to study the pathogenic driver genes of lung adenocarcinoma and explore new therapeutic target genes of lung adenocarcinoma. [0003]...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N5/10C12N15/54
CPCC12N5/0693C12N9/1088C12N15/86C12N2740/15043C12Y205/01018
Inventor 宋鑫曾宝真葛春蕾孟旭东
Owner 信雅生物科技(苏州)有限公司
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