A Preliminary Study On STAT3 And Relevant CeRNA Expression And Regulation Mechanism In ICC

PART 1 A preliminary Study on STAT3 and relevant ce RNA expression and regulation mechanism in ICC ObjectiveThe purpose of this study was to verify Signal Transducer and Activator of Transcription3(STAT3)expression in intrahepatic cholangiocarcinoma tumor,adjacent mucosaand and lymph gland,to verify the different expression level of them.We use ce RNA data base to search candidate for Competitive endogenous RNA of STAT3,RT-PCR detect ce RNA affect expression level of STAT3 and explore regulatory mechanism.BackgroundSignal Transducer and Activator of Transcription3 is a kind of cancer genes which widelyexpressed in various tissues and cells.Kinase stimu Lating STAT3 by phosphorylation,it will express itsbiology message to cell nucleus by cytokines and growth factors,in that,it will restrain effect transcription of target gene,such as promote cell proliferation and promote or inhibit microvascu Lar generation and cell adhesion.Signal Transducer and Activator of Transcription3 is a part of signal transduction pathway of JAK-STAT.The hypothesis of competing endogenous RNAs expresses a theory that RNAs from different kind can effect others’ expression by combine others’ MREs through the same micro RNA.This competition between RNAs makes a great influence of RNAs’ regu Lation and control and it also makes a big NET of regu Lation and control between ce RNAs.Morbidity of Intrahepatic cholangiocarcinoma is10%-15% account for malignant primary hepatic tumor,it is the second place after hepatocellu Lar carcinoma.Intrahepatic cholangiocarcinoma(ICC)stem from the epithelial cell of hepatic secondary bile duct or branch of secondary bile duct.ICC included many kinds of pathology,most of ICC is adenocarcinoma with diverse differentiation.Cells of ICC infiltrative growth in hepatic portal areas,those cells invasivese blood vessels and nerves.This kind of invasive growth make ICC easier invasivese adjacent organs and lymph,and it makes cure rate and resection rate low than other tumor.Up to now,it is widely accepted by all over the world the most effective therapy method is surgical operation(R0),but surgical operation do not prolong patients’ life in 5 years or prolong patients’ progression free survival.And unfortunately,most of ICC patients have no chance to accept surgical operation,because the type of invasive growth of ICC and have no obvious symptoms in early stage of ICC.In recent research find the expression of Signal Transducer and Activator of Transcription3 is proved effected prognosis of many types of tumor,such as acute myelogeinous leukemia and cutaneum carcinoma and carcinoma of non-small cell lung.As above the research of the pathogenesis tumors show that micro RNA can be direct or indirect participation STAT3 activation regulation in tumor.Hypothesis,according to the competitive endogenous RNA,ce RNA through the common mircro RNA binding sites combine with its competitive,this reminder us micro RNA can connect ce RNA with STAT3 as an intermediary.Base on consult literature,we found that ce RNA regulate the expression of STAT3 in intrahepatic bile duct carcinoma undefined,which laid a theoretical basis for our research.Methodpathological scoresWe use immunohistochemical to dyeing tumor tissue and para-carcinoma tissue and lymph gland,according to consequence of dyeing we grade expression of Signal Transducer and Activator of Transcription3(STAT3)level.functional test of STAT3We use pc DNA3.1 as carrier to build Signal Transducer and Activator of Transcription3(STAT3)overexpression plasmid,also we put pc DNA3.1-STAT3 overexpression plasmid and pc DNA3.1 carrier into RBE cell line.After that,scarification test and transwell test to confirm capacity of invasiveness.Using ce RNAs database to find candidate of STAT3’s ce RNAs,like ACSL4 and IFNAR2 and MAP3K5 and MAP3K9 and SMAD5 and SOD2 and TNFRSF10 B and XIAP and Lnc RNA-NEAT1,RT-PCR is tested the relationship between those ce RNA candidates and Signal Transducer and Activator of Transcription3(STAT3)overexpression.We use Lnc RNA-NEAT1 as candidates to explore regulatory mechanism of ce RNA regulate STAT3 expression in ICC.ConclusionSTAT3 in ICC tumor express high-level than adjacent mucosaand of ICC.Signal Transducer and Activator of Transcription3(STAT3)can promote tumor metastasis.RT-PCR reveal Lnc RNA-NEAT1 have significant positive correlation with expression level of STAT3.Lnc RNA-NEAT1 as competitive endogenous RNA can regulate and control STAT3 in ICC.PART2 Capture and detection technology of CTC in HCC patients peripheral blood ObjectiveCircu Lating tumor cell is kind of malignant cell which diameter bigger than normal cells in blood.We use membrane with olfactory foramina to capture CTCs by characteristic of circu Lating tumor cells.Counting CTCs after eliminate interruptions of normal blood cells,comprehensive analysis the number of CTCs with clinical date(date of laboratory examination and pathogenic data)to identify CTCs affect tumour formation and pathological examination in HCC patients.BackgroundFirst announcing the concept of circu Lating tumor cell was in 140 years ago,it means malignant tumor cells in peripheral blood.In last 10 years,with progress of identification and capture method of circu Lating tumor cell is explored by a series of systematic study.Recently study have proved circu Lating tumor cells play a critical role of process in recurrence and metastasis of the tumors.Circu Lating tumor cell as a tumor biological marker detection is widely used in breast cancer,lung cancer,prostatic cancer,colon cancer,we use CTCs to inspect chemotherapeutics mu Ltidrug resistance in those cancers.Circu Lating tumor microemboli(CTM)is consists of 3 or more single circu Lating tumor cells,CTM has be confirm played a critical role in metastasis of prostatic cancer and pancreatic cancer of end-stage.People with HBV infection history,middle aged male,menopausal females,it is high risk of HCC prevalence rate.HCC patients have a unfavourable prognosis because of have no obvious symptom in early stage cause low operative resection rate.The morbidity of HCC is first place in primary hepatic carcinoma,and morbidity of HCC in eastern coastal area of China is higher than other China area.According to statistics,male is easier to get HCC than female(morbidity of HCC is 4.15:1)in mainland of china,in 2012 according to the date of WHO China’s morbidity of HCC is 52.7% to global rate.In last decades years,morbidity of HCC is getting lower by abdominal u Ltrasonic testing and injection of HBV vaccine in newborn.But total mortality of HCC till high,cause characteristic of HCC and other variety of objective factors.Main study point of carcinoma in global medical field is mechanism of tumor’s occurrence and development,metastasis and recurrence,with the development of biological technique,like high-throughput sequencing popular direction,the study of metastasis and recurrence has became popu Lar direction gradually.We use CTCs to study mechanism of metastasis and recurrence of it by characteristic of HCC.MethodOur study use YZY-CTC-BIOPSY made by Wu Han YZY biotech company as experimental platform,distinguishing CTCs and normal cells by pathomorphism and fluorescence staining.Counting CTCs after eliminate interruptions of normal blood cells,comprehensive analysis the number of CTCs with clinical date(date of laboratory examination and pathogenic date)to identify CTCs affect tumour formation and pathological examination in HCC patients.We use WGA ampli-Kit made by Taiwan Shi Xiang biotech company to amplify CTDNAs,in that,preparing for high-throughput sequencing.ConclusionIn our study we use YZY-CTC-BIOPSY instrument to detect circu Lating tumor cell and circu Lating tumor microemboli(CTM),the result reveal that CTC and CTM can be captured by our apparatus.Immunofluorescent staining can identify CTC and WBC that eliminate interference.We use laser beam cutting instrument to obtain CTC on the membrane.After statistical analysis,we find tumor capsule and tumor vascular invasion affect CTC in peripheral blood.We use QC Ampli Kit can magnify whole gene in CTC.