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Focal adhesion kinase splice isoforms and their applications

A technology of focal adhesion kinase and isomer, applied in the field of medicine

Active Publication Date: 2021-10-26
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, unlike classic lung cancer therapeutic targets such as EGFR and ALK, there are currently no studies on FAK splice isoforms in SCLC.

Method used

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  • Focal adhesion kinase splice isoforms and their applications
  • Focal adhesion kinase splice isoforms and their applications
  • Focal adhesion kinase splice isoforms and their applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] In this example, small cell lung cancer cell lines were cultured, including LCO217 (PDC cells from patients with small cell lung cancer), SBC-2 (purchased from Beina Biotechnology), H69, H82, H446, H2227, DMS153 (purchased from National Experimental Cell Resources Shared platform), H524 (Kunming Cell Bank, Chinese Academy of Sciences), H1688 (Shanghai Institute of Biological Sciences, Chinese Academy of Sciences), use Trizol method to extract RNA from small cell lung cancer cell lines, reverse transcribe cDNA, use FAK 6 / 7 Specific PCR primers were used for PCR, and the PCR products were run on gel for identification. For the results, see Figure 1A . Small cell lung cancer cell line cDNA, using FAK 6 / 7 Specific sequencing primers were used for PCR, the PCR products were subjected to Sanger sequencing, and the sequencing was performed using Snapgene software for comparative analysis. For the results, see Figure 1B .

[0030] Download the result file (CCLE_RNAseq_Exon...

Embodiment 2

[0036] This example studies the role of the expression of FAK splicing body in the clinical diagnosis of small cell lung cancer. The lung cancer samples of small cell lung cancer were selected, and the expression level of FAK splicing body was detected by RNA in situ hybridization. The specific detection method is as follows:

[0037] 1) Dewaxing: Preheat in an oven at 72°C for 2.5 hours, then put them in fresh xylene I&II&III for 10 minutes each;

[0038] 2) Hydration: 2 minutes each in absolute ethanol I & II, air-dry at room temperature, use within 24 hours or proceed directly to the next step;

[0039] 3) Hydrogen peroxide treatment: Add 5-8 drops of hydrogen peroxide on the slide, room temperature for 10 minutes, rinse the slide in ddH2O for 3-5 times;

[0040] 4) Restoration: Boil the 1X restoration solution over 100°, slowly put the slide into the repair solution, cover with aluminum foil, and repair for 15 minutes; immediately put the slide in ddH2O and rinse for 3-5...

Embodiment 3

[0069] In order to further evaluate the role of FAK splicing body in the treatment of small cell lung cancer, the present invention utilizes different FAK splicing isoforms (H82:FAK 6,7 , H446:FAK 6,7 , H69:FAK 7 , LCO217:FAK WT ) small cell lung cancer cell line, treated with different concentrations (μM) of FAK inhibitor PF-562271 inhibitor for 48 hours, CCK8 detected cell viability, the results can be found in Figure 3A .

[0070] FAK wt After the cell line LCO217 was treated with different concentrations of FAK inhibitor PF-562271 inhibitor for 48 hours, the cell pellet was collected, and the anti-p-FAK and FAK antibodies were analyzed by Western blot. The results can be found in Figure 3B .

[0071] FAK 6,7 After the cell line H82 was treated with different concentrations of FAK inhibitor PF-562271 inhibitor for 48 hours, the cell pellet was collected and analyzed by Western blot with anti-p-FAK and FAK antibodies. For the results, see Figure 3C .

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Abstract

The present invention provides focal adhesion kinase splicing isomers and applications thereof, specifically, the present invention provides the application of reagents for detecting focal adhesion kinase splicing isomers in the preparation of diagnostic agents for diagnosing small cell lung cancer, and also provides The application of focal adhesion kinase splicing isoforms as targets in screening and / or preparing drugs for the treatment of small cell lung cancer. The present invention finds that the FAK focal adhesion kinase splicing isoform can be used as a clinical diagnostic marker for small cell lung cancer, and has a good application prospect in guiding the clinical application of FAK kinase inhibitors.

Description

technical field [0001] The present invention relates to focal adhesion kinase (focal adhesion kinase, FAK) splicing isoforms and applications thereof, in particular to a detection method for focal adhesion kinase (FAK) splicing isoforms in the diagnosis of small cell lung cancer and as an anticancer drug target The new application belongs to the field of medicine. Background technique [0002] There are 2.1 million new lung cancer cases and 1.8 million lung cancer deaths every year in the world, while there are 787,000 new lung cancer cases and 631,000 lung cancer deaths in my country every year. Lung cancer is the cancer with the highest morbidity and mortality among all cancers. In terms of tissue types, lung cancer is mainly divided into two types: non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). SCLC is a poorly differentiated, high-grade neuroendocrine malignancy that accounts for approximately 13% of all newly diagnosed lung cancers. In terms of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/574G01N33/573A61K45/00A61P35/00
CPCC12Q1/6886G01N33/57423G01N33/57484G01N33/573A61K45/00A61P35/00C12Q2600/136C12Q2600/158G01N2333/912
Inventor 周光飚王桂珍解达伟孙贝贝王峥
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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