Screening method of esophagus cancer markers by tea polyphenol in tea and golden camellia

A screening method, technology of tea polyphenols, applied in the field of screening tea polyphenols in tea and Camellia japonica for markers of esophageal cancer

Inactive Publication Date: 2015-03-25
程金生
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] At present, there is no literature related to the molecular biology or epidemiology of tea polyphenols in plants such as tea (including black tea, green tea, oolong tea, camellia, etc.), Camellia japonica and other plants in inhibiting esophageal cancer.

Method used

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  • Screening method of esophagus cancer markers by tea polyphenol in tea and golden camellia
  • Screening method of esophagus cancer markers by tea polyphenol in tea and golden camellia
  • Screening method of esophagus cancer markers by tea polyphenol in tea and golden camellia

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Embodiment 1

[0036] Extraction and separation of tea polyphenols from Meizhou Dancong tea: The XDA macroporous resin adsorption purification method is used to extract and prepare tea polyphenols from Meizhou Dancong tea. Soak in 30% ethanol for 20min→microwave extraction for 20min→separate the extract through a hollow fiber membrane (Mr=10000)→concentrate by rotary evaporation→pass through a macroporous resin XDA-200 column→elution with water→elution with 10% ethanol→30 Gradient elution with % ethanol → the eluent was concentrated by rotary evaporation → freeze-drying → dry powder was obtained. The content of tea polyphenols is determined according to the method of "GB / T 8313-2008 Determination of the content of tea polyphenols and catechins in tea leaves", and the content of tea polyphenols = (mass of tea polyphenols in sample powder / mass of sample) × 100%, the tea polyphenol separation and purification method described in the present invention, the tea polyphenol content is above 85%.

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Embodiment 2

[0048] Extraction and separation of tea polyphenols in Camellia japonica: XDA macroporous resin adsorption purification method to extract and prepare tea polyphenols from Camellia japonica, the main steps include: weighing the dried flowers of Camellia japonica → crushing through a 30-mesh sieve → soaking in 30% ethanol for 20 minutes → Microwave extraction for 20min → Separation of extract by hollow fiber membrane (Mr=10000) → Concentration by rotary evaporation → Passing through macroporous resin XDA-200 column → Water elution to remove impurities → 10% ethanol elution to remove impurities → 30% ethanol gradient elution → Concentrate the eluate by rotary evaporation → freeze-dry → obtain a dry powder. The content of tea polyphenols is determined according to the method of "GB / T 8313-2008 Determination of the content of tea polyphenols and catechins in tea leaves", and the content of tea polyphenols = (mass of tea polyphenols in sample powder / mass of sample) × 100%, the tea p...

Embodiment 3

[0060] Extraction and separation of tea polyphenols from Dapu Xiyan camellia: XDA macroporous resin adsorption purification method is used to extract and prepare tea polyphenols from Dapu Xiyan camellia. The main steps include: Dapu Xiyan camellia is preferably weighed → crushed to 30 mesh Sieve→soak in 30% ethanol for 20min→microwave extraction for 20min→separate the extract through a hollow fiber membrane (Mr=10000)→concentrate by rotary evaporation→pass through a macroporous resin XDA-200 column→elution with water→elution with 10% ethanol Impurities → 30% ethanol gradient elution → eluents were concentrated by rotary evaporation → freeze-drying → dry powder was obtained. The content of tea polyphenols is determined according to the method of "GB / T 8313-2008 Determination of the content of tea polyphenols and catechins in tea leaves", and the content of tea polyphenols = (mass of tea polyphenols in sample powder / mass of sample) × 100%, the tea polyphenol separation and purif...

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Abstract

The invention discloses a screening method of esophagus cancer markers by tea polyphenol in tea and golden camellia. The tea polyphenol in tea and golden camellia is extracted and prepared from different types of tea and golden camellia, and the tea polyphenol acts on human esophageal squamous cells and adenocarcinoma cell OE33 which are cultured in vitro; an anti-proliferative effect on esophagus cancer by the tea polyphenol is detected in an MTT method, and an inducing differentiation effect on the esophagus cancer by the tea polyphenol is detected by flow cytometry so as to observe cell cycles and apoptotic effects; cell morphological change is observed by using a fluorescence inversion microscope; protein expression change in the esophagus cancer cells is analyzed by means of immunohisochemistry, Real-time fluorescent quantitation RT-PCR and Westem-blot. Based on SPSS1 statistics, the detection results are analyzed, and differential expression proteins obtained by performing action on the esophagus cancer by the tea polyphenol in the tea and golden camellia are detected, and associated proteins are screened out according to database search and matching. In the invention, a mechanism of the function of tea polyphenol can be expounded in accordance with the research on marker molecules of the differential expression proteins acted by the tea polyphenol.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a screening method for markers of esophageal cancer by tea polyphenols in tea and camellia japonica. Background technique [0002] Tea polyphenols (tea polyphenols, TP), also known as tea tannins and tea tannins, are the general term for polyphenols in tea, including flavanols (catechins), anthocyanins (anthocyanins and anthocyanins) ), flavonoids, flavonols, phenolic acids and phenolic acids, etc. Tea polyphenols are a class of chemical substances containing polyphenolic hydroxyl groups, which can scavenge active free radicals in the body and have strong antioxidant effects, so they are a promising natural food antioxidant. BHT (Antioxidant 264), BHA (Butylated Hydroxyanisole), etc., although they have many advantages, they also have different degrees of toxicity. As a natural antioxidant, TP not only has the advantages of high safety and no side effects, but also has better antio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02C12Q1/68G01N33/68G01N33/573
Inventor 程金生朱文娟郑启祥韦卓恒陈信炎
Owner 程金生
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