Clinic Correlation Between MiR-22/TET2 And The Development Of HCC Patients With Chronic Alcohol Consumption

Backgrounds:In recent years,cancer has become a major killer which threaten the health of human..And in worldwide,liver cancer is a malignant tumor with higher morbidity and mortality,besides China is the high-prevalence area.and for the past few years liver cancer showed a trend of high incidence and earlier age.But the mechanisms and the etiology of liver cancer are still unclear.we know liver cancer Is related with several causes and regulated by multipal mechanisms.The research show that the pathogenesis of liver cancer are associated with virus infection,cirrhosis,alcohol,aflatoxin,water pollution,nitrosamines substances and trace elements.what our team study is the relationship between alcohol and the development and metastasis of liver cancer.Not only alcohol can increase the risk of hypertension,apoplexy and heart disease,but also be a risk factor of liver cancer,colorectal cancer,esophageal cancer,breast cancer and other cancers.In the European and American countries,alcohol is the largest cause of liver cancer.In China,alcohol is second to the viral hepatitis which cause the liver cancer and has an increasing trend,Overall,light drinking was associated with a small but non-significant increased risk of cancer in both women and men,Increasing consumption of alcohol also increase the risk of cancer gradually.So heavy drinking is a dangerous cause for cancer.But the mechanism how different ahlcohol consumption affect HCC progression and angiogenesis is little known.Our team has discovered that chronic alcohol consumption can promote the progression and metastasis through NF-?B signaling.then we hypothesis other signaling may participate in the regulation.Gene microarray show us that mi R-22 increased evidently.The literature luciferase reporter gene display TET2 is mi R-22 of downstream target genes.My research involved if TET2 affect progression and metastasis of the solid tumor HCC.TET2 comes from TET family which involve three member TET1,TET2,TET3,TET proteins play an important role in the regulation of DNA methylation,which participate in DNA demethylation by iteratively oxidizing 5-methylcytosine(5m C)to 5-hydroxymethylcytosine(5hm C),5-formylcytosine(5f C),and 5-carboxylcytosine(5ca C)Research has proved thataberrations in DNA methylation patterns are observed from the early stages of carcinogenesis.Recently change of TET expression is related to not only cell pluripotency,but also the occurrence of cancer TET2 is situated in the 4q24 region The study found that the TET2 gene mutations are widely exist in the blood malignant tumor(such as AML CMML,MPS),recent years scientists devote to study if the mutation occurs in the solid tumor and found TET2 mutation in some solid cancer(breast cancer and lung cancer)is chronic alcohol consumption a risk factor for the HCC?c an alcohol change the expression of TET2?what is the relationship between TET2 and the progression and metastasis.To test this hypothesis,Firstly we collected clinical data from hospital and the analysis found that the HCC patients with chronic alcohol consumption have a high rate of vessel metastasis and a low rate of survival comparing to patients without alcohol consumption.It confirmed that alcohol can promote the development and metastasis.Then through immunohistochemical we found TET2 expression in HCC with chronic alcohol organizations is significantly lower than that without alcohol consumption To validate TET2 effects in liver cancer in vitro further.We transferlsi RNATET2 into liver cancer cells to knock down TET2 expression,,and then observe the proliferation,cell cycle and migration of liver cancer cell.Research contents:1We collected the stastistis of HCC and divided the patients into two groups,the HCC with chronic alcohol consumption and the HCC without alcohol consumption.Then we discovered the difference in prognosis and vessal metastasis between the two groups.2Collect the pathological tissue section and detect the TET2 expression in HCC tissues via IHC.Then we compare the TET2 expression of two groups3 transfer si RNA-TET2 into liver cancer cells to knock down TET2 expression,and get a cell line with lower TET2 expression.4Study the cell biology function of cells after transferring si RNAA detect Hep G2 and 7721 cells proliferation via MTT and colony formation assayB detect Hep G2 and 7721 cells migration and invasion abilities via transwell assay.C observe the capacity of anchor-independent growth.via Soft agar semisolid colony forming assayRearch results1 From the stastistis analysized by SPSS we found that HCC with alcohol consumption have a higher rate of vessel metastasis and a lower rate of survival comparing to patients without alcohol consumption2 IHC detected that TET2 expression in HCC with alcohol consumption is lower than patients without alcohol consumption.3 Influence of TET2 on cell biology of liver cancer cells.A the influence of TET2 on the Hep G2 and 7721 cells growth: MTT and colony formation assay results detected that the proliferation capability of Hep G2 and 7721 cells was increased after knocking down the expression of TET2B the influence of TET2 on the Hep G2 and 7721 cells migration and invasion abilities:transwell assay results detected that the migration and invasion capability of Hep G2and 7721 cells was increased after knocking down the expression of TET2C the influence of TET2 on the Hep G2 and 7721 cells anchor-independent growth: Soft agar clone forming assay results displayed that the ability of anchor-independent growth of of Hep G2 and 7721 cells was increased after knocking down the expression of TET2Conclusion1that HCC with alcohol consumption have a higher rate of vessel metastasis and a lower rate of survival comparing to patients without alcohol consumption2 the alcohol can affect the expression of TET23 Inhibition of TET2 increases proliferation,migration,and invasion of liver cancer cells in vitro.TET2 plays the antioncogene role in liver cancer cells.