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Prevention of lung cancer target plekhn1 and its application

A technology of PLEKHN1, 1.PLEKHN1, applied in the new target field of lung cancer, can solve the problems of increased lung cancer, incompletely clear mechanism of lung cancer, resistance, etc., and achieve the effect of preventing lung cancer and promoting the malignant transformation of lung cells

Active Publication Date: 2021-09-03
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since arsenide is not only an important environmental carcinogenic compound, but also an important model compound for laboratory-induced lung cancer, the study of its carcinogenic mechanism may be of great significance to the biological effects of arsenic (ie, arsenic-induced lung cancer) and the general mechanism of lung cancer. However, the specific mechanism of arsenic-induced lung cancer is not completely clear, which undoubtedly adds great resistance to our prevention of lung cancer, especially arsenic-induced lung cancer

Method used

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  • Prevention of lung cancer target plekhn1 and its application
  • Prevention of lung cancer target plekhn1 and its application
  • Prevention of lung cancer target plekhn1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Normal lung epithelial cells were treated with arsenic to up-regulate the protein expression level of PLEKHN1 in a concentration- and time-dependent manner

[0047] 1 Cell poisoning treatment

[0048] (1) Short-term exposure of cells to arsenic poisoning:

[0049] a) Multiple wells of a six-well plate for cell seeding, 200,000 Beas-2B cells per well, using DMEM medium containing 10% FBS, at 37°C in constant temperature CO 2 Cultivate in an incubator; the cell density reaches about 70% after 24 hours.

[0050] b) 24 hours after seeding the plate, start to replace the medium with DMEM medium containing 0.1% FBS, and treat for 12 hours according to the same culture conditions as above.

[0051] c) After 12 hours, add arsenic compounds with concentrations of control (0), 0.5, 1 and 2 μM respectively for 12 hours ( figure 1 A); the fixed arsenic concentration was 1 μM, and the control group was treated (0), 6, 12 and 24h ( figure 1 B). The result is as figur...

Embodiment 2

[0098]Example 2. The expression level of PLEKHN1 in clinical lung cancer tissues shows an overall up-regulation trend.

[0099] 1. TCGA database data analysis

[0100] Download lung cancer sequencing data and clinical data from the TCGA database (https: / / tcga-data.nci.nih.gov / ). Use the R package edgeR to preprocess the data, standardize the raw count to log-CPM value, perform linear modeling, and use the precision weight calculated by the voom function to adjust the average variance relationship. Using the linear regression and empirical Bayesian methods provided by the limma package, the differential expression analysis of Tumor VS Normal in the mRNA data was performed. After statistical calculation, the corresponding P.Value value was obtained, and the Benjamini&Hochberg method was used for multiple test correction to obtain the corrected P value, which is adj.P.Value. Differential expression thresholds are all adj.P.Value2. The result is as figure 2 described in A. T...

Embodiment 3

[0129] Example 3, PLEKHN1 can significantly promote the formation of Beas-2B cell clone colonies induced by arsenide for a long time.

[0130] 1 Cell transfection related steps (1) Lipofectamine transfection method:

[0131] a) Beas-2B cells were plated for 24 hours, and when the cells grew to 80%, 1 ml of full culture was replaced 1 hour in advance.

[0132] b) Add 1 μg of target DNA (GFP-PLEKHN1 plasmid or PEGFP-C1 plasmid, both purchased from Shanghai Sunny Biotechnology Co., Ltd.) into 50 μl of serum-free DMEM containing high glucose and mix well.

[0133] c) Take 3μl PolyJet TM Add 50 μl of serum-free DMEM containing high glucose to the reagent, and mix gently with a gun for 3-4 times.

[0134] d) Immediately place the diluted PolyJet TM Add the reagent to the diluted DNA solution, and gently suck up and down to mix 3-4 times. (sequence only PolyJet TM plus DNA)

[0135] e) Incubate at room temperature for 10-15 minutes, not exceeding 20 minutes.

[0136] f) Add 1...

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Abstract

The invention belongs to the field of biotechnology, and specifically relates to PLEKHN1 as a new target for preventing and diagnosing arsenic-induced lung cancer and its application. Specifically, the present invention discloses the application of PLEKHN1 expression inhibitors in the preparation of drugs for the prevention / treatment of lung cancer; knocking down PLEKHN1 can significantly inhibit the malignant transformation of normal lung cells induced by arsenic compounds.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to PLEKHN1 as a new target for preventing and diagnosing arsenic-induced lung cancer and its application. Background technique [0002] According to the 2018 global cancer analysis data, the incidence of lung cancer accounts for 11.6% of all cancers, while the mortality rate accounts for 18.4% of all cancers. Its morbidity and mortality rates rank first among all major cancers. According to the data of cancer incidence in China in 2015, the incidence of lung cancer reached 20.03%, and the mortality rate was as high as 26.99%. Therefore, the prevention and treatment of lung cancer is urgent for China and the world. Arsenic is a non-metallic element that widely exists in nature and is widely used in people's production and life, including pesticides, insecticides, herbicides and alloys. However, arsenic and its compounds are like a double-edged sword. While we are benefiting...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11G01N33/574A61K45/00A61P35/00
CPCA61K45/00A61P35/00C12Q1/6886C12Q2600/136C12Q2600/158G01N33/57423G01N33/57484
Inventor 黄海山林炜炜金红蕾赵玲玲
Owner WENZHOU MEDICAL UNIV
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