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Application of STAT3 (signal transducer and activator of transcription 3) inhibitor-EGCG (epigallocatechin gallate) in colon cancer treatment

An inhibitor, colon cancer technology, applied in the application field of STAT3 inhibitor-EGCG in the treatment of colon cancer

Inactive Publication Date: 2014-06-04
SHANGHAI MCARY BIOMEDICINE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(3) RNA interference: RNA interference (RNA interference, RNAi) is a post-transcriptional gene silencing mechanism (Postoranscriptional gene silencing, PTGS) triggered by double-stranded RNA (double stranded RNA, dsRNA). Under the action of (dsRNA-specific endonuclease), a small interfering RNA (small interfering RNA, siRNA) with a size of 22 bp can be formed, and the siRNA can be further incorporated into a multicomponent nuclease (multicomponent nuclease, KISC) and activated to thereby Precisely degrade mRNA with the same sequence as siRNA, completely inhibiting the translation and expression of the gene in cells. Since there is no suitable transgenic technology at present, the application of RNAi in clinical treatment needs further research

Method used

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  • Application of STAT3 (signal transducer and activator of transcription 3) inhibitor-EGCG (epigallocatechin gallate) in colon cancer treatment
  • Application of STAT3 (signal transducer and activator of transcription 3) inhibitor-EGCG (epigallocatechin gallate) in colon cancer treatment
  • Application of STAT3 (signal transducer and activator of transcription 3) inhibitor-EGCG (epigallocatechin gallate) in colon cancer treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Embodiment 1: MTT assay measures the influence of EGCG on BEL-7402 cell proliferation

[0013] Set up the control group and the EGCG group (10, 20, 40, 80, 160, 320Hmol / LEGCG), take the cells in the logarithmic growth phase, digest them with DMEM containing 10% fetal bovine serum to make single cells Suspension, inoculated in 96-well culture plate, 100 μL per well (containing about 5*10 3 After the cells adhered to the wall, the culture solution was removed and drugs were added. Three replicate wells were set up in each group, and three zero-adjustment wells were set up (no cells were added, only 100 μL culture solution was added), and the culture was continued for 48 h. Aspirate off the medium and wash it twice with 150 μL of PBS, then add 10 μL of MTT to each well, stop the culture after 4 hours, discard the supernatant, add 150 μL of lysate to each well, shake on a horizontal shaker for 10 min, and place in a microplate reader (Tecan F200 ) to measure the absorbance...

Embodiment 2

[0014] Example 2: Flow cytometry detection of HCT-116 cell apoptosis induced by EGCG

[0015] Cells in the logarithmic growth phase were taken and cultured in DMEM containing 10% fetal bovine serum after routine trypsinization.

[0016] The culture solution was made into a single cell suspension, counted and planted in a six-well plate, each well containing about 2*10 5 After the cells adhered to the wall, remove the culture medium and add drugs, add 2 mL of 0, 40, 80, 160, 320 pmol / L EGCG culture solution respectively, culture for 48 hours, digest and centrifuge the cells with EDTA-free trypsin , wash the cells twice with cold PBS. Then use 400 μL 1×Annexin V to bind the suspended cells, add 5 μL Annexin V-FITC staining solution to the cell suspension, mix gently and incubate in the dark for 15 minutes, add 10 μL PI staining solution and incubate in the dark for 5 minutes, and use flow-through within 1 hour Cytometer (Accuri C6) detection.

Embodiment 3

[0017] Example 3: PCR technology detects the inhibitory effect of EGCG on the susceptibility gene of HCT-116. Take the cells in the logarithmic growth phase, digest them with 10% fetal bovine serum DM EM culture medium to make a single cell suspension, and count Seed in a six-well plate, each well contains about 2*10 5 After the cells adhere to the wall, remove the culture medium and add drugs. Add 2 mL of 20, 40, 80, and 160 μmol / LEGCG culture solution respectively. After culturing for 48 hours, trypsinization without EDTA was used to collect the cells by centrifugation, and the cells were washed twice with cold PBS. HCT-116 cells were collected, lysed, and total RNA was extracted, cDNA was synthesized by reverse transcription with the kit, and then amplified by PCR. Use a spectrophotometer to measure the amplified RNA, and take 2 μg of total RNA for agarose gel electrophoresis

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Abstract

The abnormal adjustment and control of an Stat3 (signal transducer and activator of transcription 3) signal transduction system generally exist in canceration cell strains. The invention provides a compound EGCG (epigallocatechin gallate) which is an effective Stat 3 inhibitor. The Stat 3 inhibitor can be applied to preparation of cancer therapy medicines containing endogenous activated Stat3 signal paths.

Description

technical field [0001] The invention relates to the application of a signal transduction system inhibitor compound in the preparation of a drug for treating colon cancer. Background technique [0002] According to WHO statistics, more than 11 million cases of malignant tumors occur every year in the world, and more than 8 million people die from them. The annual incidence of tumors in developed countries is higher than 300 / 100,000. In the early days of the founding of New China, malignant tumors only accounted for the 9th to 10th cause of death, and rose to the third place in the 1970s. According to national health data statistics, in the past two years, malignant tumors accounted for the first cause of death among urban residents in my country, and there were about 2.2 million new patients every year. WHO released the latest "World Cancer Report" in Geneva not long ago, saying that by 2020, the global cancer incidence may increase by more than 50% compared with the present...

Claims

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Application Information

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IPC IPC(8): A61K31/353A61P35/00A61P1/00
Inventor 徐学军魏洁麟
Owner SHANGHAI MCARY BIOMEDICINE TECH
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