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Application of estrogen-related receptor alpha as diagnostic marker for cutaneous squamous cell careinoma and related application of application

A technique for squamous cell carcinoma and diagnostic marker, applied in the fields of molecular biology and tumor prevention and treatment, to reduce recurrence and improve survival rate

Inactive Publication Date: 2017-08-18
复旦大学附属华山医院北院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The treatment of squamous cell carcinoma is currently available with local drug therapy, excision, electrodrying, radiation therapy, etc., but the disadvantages of these treatments still exist

Method used

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  • Application of estrogen-related receptor alpha as diagnostic marker for cutaneous squamous cell careinoma and related application of application
  • Application of estrogen-related receptor alpha as diagnostic marker for cutaneous squamous cell careinoma and related application of application
  • Application of estrogen-related receptor alpha as diagnostic marker for cutaneous squamous cell careinoma and related application of application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The fluorescence quantitative PCR experiment was used to test the transcriptional regulatory activity of XCT790 and ERRαshRNA on ERRα in A431 (human skin squamous carcinoma cells).

[0033] This example illustrates that XCT790 and ERRα shRNA involved in the present invention can effectively inhibit the transcription of ERRα gene in human skin squamous carcinoma cell A431, indicating 1) ERRα mRNA is expressed in human skin squamous carcinoma cells, and the expression level is higher than normal Skin cells; 2) Using XCT790 or ERRαshRNA significantly down-regulated the level of ERRαmRNA in A431. Fluorescence quantitative PCR testing technology is a technology familiar to workers in the field.

[0034] 1. Cell plating:

[0035] Warm the DMEM cell culture medium and pancreatin with 10% FBS at 37℃ in advance, wipe the sterile operation table with alcohol for sterilization, wipe the pancreatin and DMEM cell culture medium with alcohol and put it into the sterile operation table, and...

Embodiment 2

[0067] Western Bloting was used to test the effect of XCT790 and ERRαshRNA on the expression level of ERRα protein in A431 (human skin squamous carcinoma cells). Western Bloting cell proliferation experiment is a technique familiar to workers in the field.

[0068] 1. Extract the total protein of A431 cells

[0069] After XCT790 or ERRαshRNA intervenes for the corresponding time, aspirate the old culture medium in the original cell culture class, wash the cells 2-3 times with pre-cooled PBS, and add 200μL of cell RIPA lysis solution to each well of the 6-well plate (before use A few minutes ago, PMSF was added to the final concentration of 1μM), and the lysate was in full contact with the cells after a few blows with a gun. After 1-2s, the cells are fully lysed. After sufficient lysis, collect the cells with a cell scraper on ice. Use a pipette to transfer the lysate into a 1.5 mL centrifuge tube, mix by pipetting, and place on ice for 30 minutes. Centrifuge at 12000 rpm at 4°C...

Embodiment 3

[0099] The CCK-8 cell proliferation experiment was used to determine the proliferation inhibitory effect of XCT790 and ERRαshRNA on A431 (human squamous carcinoma cells).

[0100] This example illustrates that the ERRα involved in the present invention can effectively inhibit the proliferation of human skin squamous carcinoma cell line A431 after being down-regulated by the specific inverse agonist XCT790 or shRNA. The proliferation of cancer cell lines in vitro shows extremely important key effects. The CCK-8 cell proliferation experiment is a technique familiar to workers in the field.

[0101] 1. A431 cell culture and seeding 96-well cell culture plate:

[0102] Take out the A431 in good logarithmic growth phase and observe the cell morphology, if the cells have been 80% confluent, start to passaging. Wash the old culture medium in the petri dish, wash 3-5 times with sterile 1mL PBS, add 1 mL of warm-bathed pancreatin, shake the petri dish horizontally to make the pancreatin eve...

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Abstract

The invention discloses an application of an estrogen-related receptor alpha (ERRalpha) as a diagnostic marker for cutaneous squamous cell careinoma and a related application of the application. The invention discloses an application of the ERRalpha and an active component for lowering or blocking molecular expression of the ERRalpha in cutaneous squamous cell careinoma in preparation of related kits and drugs for diagnosing and treating cutaneous squamous cell careinoma. Novel diagnosis and treatment means are provided for treatment of cutaneous squamous cell careinoma.

Description

Technical field [0001] The present invention relates to the technical fields of molecular biology and tumor prevention and treatment, in particular to the use of an estrogen-related receptor α as a diagnostic marker for skin squamous cell carcinoma and related applications. Background technique [0002] Skin cancer is one of the most common cancers in humans. The occurrence of skin cancer is mainly caused by the uncontrollable growth of epidermal cells. Skin cancer is divided into basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). ) And melanoma (melanoma) three types, which originate from the basal cells, squamous cells and melanocytes of the epidermis. Among them, basal cell carcinoma and squamous cell carcinoma are collectively called non-melanoma (Non-melanoma). Among non-melanoma, the proportion of basal cell carcinoma (80%) is greater than that of squamous cell carcinoma (20%). Squamous cell carcinoma is metastatic, so it is more harmful than basal cell carcinom...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/5743G01N33/57484G01N33/6872G01N2333/723
Inventor 李群益陈海飞杜鹃潘洁雯钟明康施孝金
Owner 复旦大学附属华山医院北院
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