| Background: Gastric cancer is one of the most common malignant tumor of digestive tract,according to the world health report only the latest,the incidence of gastric cancer among malignant tumors worldwide incidence of fifth,the mortality rate is ranked third,second only to lung cancer and liver cancer,he Jie reported by Professor of cancer incidence and mortality of the new me the rate of the display Chinese every year the number of new cases of gastric cancer in 400 thousand cases,accounting for the number of new cases and 42% deaths of about 300 thousand.Gastric cancer is still a major disease that endangers the health of our people.Compared with the low incidence rate in Europe and America and the higher rate of early gastric cancer diagnosis in Japan and South Korea,China still faces severe situation in combating gastric cancer.At present,the main clinical treatment of gastric cancer is resection combined with chemotherapy and radiation therapy means.Chemotherapy drugs chest(5-FU)are commonly used,in addition to paclitaxel,cisplatin and their combination.Surgical treatment,chemotherapy and radiation therapy will give patients a lot of pain,with unimaginable toxic side effects in the long term.Therefore,to find a non-toxic side effects of targeted drugs and reveal the target of drug action has a very important clinical significance for the clinical treatment of gastric cancer,improving the prognosis of patients and improving the quality of life of patients.Resveratrol is a natural polyphenolic compound,widely found in grapes,peanuts,Polygonum cuspidatum,mulberry and veratrum and other plants and food.A large number of studies have shown that resveratrol has anti-inflammatory,anti-oxidation,anti Cardiovascular,anti-aging,anti-tumor and other aspects of pharmacological effects.Therefore,as an anti-tumor drug,resveratrol has a very important and unique medicinal value.Proteomics is widely used in tumor research.The application of proteomics in the treatment of gastric cancer is mainly focused on the discovery of anti-gastric cancer drug targets and the pharmacological mechanism of certain anticancer drugs.Many studies have proved that the application of proteomics is conducive to reveal the law of life activities and also help to promote the individualized cancer patients targeted therapy,having broad prospects for development.Previous studies of resveratrol on gastric carcinoma,mainly focuses on the influence on proliferation and apoptosis of gastric cancer cells,cell cycle,study on gastric cancer cell migration and invasion mechanism,the relevant research has not been reported to study the effect of resveratrol on gastric cancer susceptibility and resveratrol on gastric cancer cell group protein interaction.Therefore,from the point of view of proteomics,the effect of resveratrol on the proteome of gastric cancer is discussed.It is necessary to find a new and effective drug target for the treatment of gastric cancer.Objective: This study is designed(1)to investigate the effect of resveratrol on the proteome of human gastric cancer cell lines with different sensitivities;(2)to explore the possible mechanism of resveratrol and the possible causes of differences in cell lines(3)to find possible anti-tumor target or tumor resistance mechanism,in order to provide the basis for carrying out research in this field,achieving drug clinical transformation and individualized treatment of cancer.Methods:(1)Cell line screening and drug treatment: select human gastric cancer cell lines with different sensitivities to resveratrol,BGC-823 and AGS,respectively culture them in the mediums with 10% homemade fetal bovine serum,1% antibiotics(penicillin and streptomycin)in DMEM and HAM’S / F12.The cells in the drug treatment group are treated with 100μM resveratrol for 48 h,and the cells and cell climbing tablets are collected.(2)Proteomics: extract proteins from the cells before and after resveratrol treatment and subject them to two-dimensional gel electrophoresis.Identify differential protein spots by mass spectrometry,and select potential anti-tumor targets.(3)Detect the expression of CK18 and E-cadherin in BGC-823 and AGS cells before and after resveratrol treatment by Real-time PCR,ICC and Western-blotting.Detected the expression of CK18 and E-cadherin in gastric tissue array by IHC and perform the correlation analysis.Results:(1)Two-dimensional electrophoresis and mass spectrometry showed that the expression of CK18 was increased after resveratrol treatment of BGC-823 and AGS cells,and the expression of CK18 was up to 7.6 times after treatment with 100μM resveratrol for 48 h.(2)The expression of CK18 in gastric cancer cell line BGC-823 and AGS was significantly up-regulated by 100μM resveratrol.The expression of E-cadherin in BGC-823 and AGS was up-regulated 48 hours after resveratrol treatment.The results of Real-time PCR showed that CK18 was slightly up-regulated in BGC-823 and slightly down-regulated in AGS after treatment with 100μM resveratrol.The expressions of E-cadherin in BGC-823 ws up and slightly down-regulated in AGS.The results of Western-bloting showed that the expression of CK18 was up-regulated in BGC-823 and AGS cell lines after 48 hours of treatment with 100μM resveratrol,and E-cadherin did not show the target band.IHC results showed that the expression of CK18 and E-cadherin in gastric tissue and pulp were statistically different and correlated with the classification of gastric tissue.CK18 and E-cadherin expressions in gastric tissue analysis and statistical analysis were significantly different and positively correlated.Conclusion:(1)After the treatment of resveratrol,the two cell lines with different sensitivities,only 8%(3/42)of the changes occurred,and said that resveratrol could affect the expression of sensitive and insensitive cell lines differently(2)The analysis results from the current protein,protein mainly concentrated in the cytoskeleton,cell adhesion,signal transduction,transcriptional regulation and metabolism related protein,suggesting that resveratrol has anti tumor biological effects of multi and multi target feature.However,the up regulation proteins of AGS are mainly concentrated in intermediate fibers,and 823 are mainly concentrated in the regulation of cell cycle.(3)Human gastric cancer cell lines BGC-823 and AGS by 100 M resveratrol treated epithelial markers CK18 and E-cadherin expression and the expression in AGS was significantly increased,suggesting that associated with resveratrol sensitivity.(4)CK18 and E-cadherin expression in gastric tissue array membrane was significantly positive correlation,negative expression of CK18 in poorly differentiated carcinoma tissues and the expression rate of E-cadherin was significantly increased,and increased with the malignant degree,the negative expression rate increased.These results suggest that the decrease in membrane expression of the two is closely related to the occurrence and development of the tumor.(5)Poorly differentiated gastric cancer cell lines BGC-823 and AGS,expression of CK18 and E-cadherin after treatment with resveratrol,suggesting that resveratrol may have to epithelial mesenchymal transformation induced by gastric cancer,and CK18 may be as resveratrol acts on the target of anti-tumor effect of gastric cancer. |
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