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The Expression of Adiponection,Adiponectin Receptors And Its Influences on HT-29Cells’ Proliferation and Migration

Author: SunHao
Tutor: TanZhiJun
School: Tianjin Medical University
Course: Surgery
Keywords: Colorectal cancer adiponectin adiponectin receptors MTTScratch-wound assay Flow cytometry immunohistochemical staining
CLC: R735.34
Type: Master's thesis
Year: 2013
Downloads: 4
Quote: 0
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Abstract


Adiponectin is adipose cytokines which is secreted by adipocyte,it was isolated firstly from the rat adipocytes by Scberer in1995,and then it was named as adiponectin by Arita in1999.Research on Adiponectin largely focused on obesity and diabetes disease.Studies were showed that Adiponectin concentration in plasma was related to abdominal fat content,changes of diponectin would be linked to abdominal obesity,insulin resistance and atberosclerosis lipid profile.Recently,marly researchers found that Adiponectin leveles were associated with some tumors.Clinical epidemiological studies indicted that the low Adiponectin levels increased the risk of colorectal cancer,breast cancer,stomach cancer and prostate cancer.As Adiponectin leveles were related to colorectal cancer,it is inferable to investigate the effects of recombinant adiponectin on the invasive and proliferation ability of human Colorectal cancer cells line HT-29in vitro,the effect of RA on proliferation of HT-29cells was measured by MTT assay;the invasive ability of HT-29cell were assayed by scratch-wound assay;the apoptosis of HT-29was measured by flow cytometric.The immunohistochemical staining was performed to detect the expressions of adiponectin receptors and adiponectin.Objective:To investigate the expression of adiponectin receptors and adiponectin in colorectal carcinoma and their relationship with clinical pathologic features;To investigate the effects of recombinant adiponectin on the invasive and proliferation ability of human Colorectal cancer cells line HT-29.Methods:HT-29cells were divided into control group and RA(2.5,5.0,10,15ug/ml) treatment groups.The effect of RA on proliferation of HT-29cells was measured by MTT assay;the invasive ability of HT-29cell were assayed by scratch-wound assay.The immunohistochemical staining was performed to detect the expressions of adiponectin receptors and adiponectin77cases of colorectal carcinoma.Results:Immunohistochemistry found that colorectal cancer tissue adiponectin receptors and adiponectin expression in normal colon tissue was significantly reduced,The expression of AdipoRl,AdipoR2and ADP was interrelated with the depth of invasion (P<0.05),the expression of AdipoRl and ADP was negative interrelated with the depth of invasion T stage (r=-0.237P=0.038, r=-0.469P=0.000); The expression of AdipoRl,AdipoR2and ADP of colorectal cancer was correlated with the lymphatic metastasis classification (P<0.05), the expression of AdipoRl and ADP was negative interrelated with the lymphatic metastasis (r=-0.237P=0.038, r=-0.231P=0.043); AdipoRl and ADP positive expression rate of T stage and lymph node metastasis were negatively correlated (r=-0.237P=0.038,r=-0.354P=0.002);The expression of AdipoRl was correlated with colorectal cancer lymph node metastasis(P<0.05); The AdipoR2and ADP expression and colorectal cancer lymph node metastasis was no significant correlation.The growth and invasive ability of HT-29cells were obviously inhibited when the concentration of RA was used(P<0.01),and this effect was dose-dependent using MTT test;Through scratch experiments,tumor cell migration was inhibited by adiponectin, which was positive correlated to adiponectin concentrations;Cell mitotic was arrested in G1phase by adiponectin,"limit point" was inhibited, and replication of cell A and chromosome was prevented.Inhibition of tumor cell proliferation by adiponectin was further confirmed;The expression of Adiponectin receptor in Colorectal cancer tissue was significant reduced,and the nuclear translocation of (3-catenin was increased significantly.Conclusion:Adiponectin receptors and adiponectin expression in colorectal cancer tissue was significantly reduced compared with normal colorectal tissue,and the low expression was significantly correlated with the invasion and metastasis of colorectal cancer,adiponectin and adiponectin receptor expression was positively correlated; adiponectin can inhibit colon cancer cell HT-29proliferation and migration.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Intestinal neoplasms > Colorectal tumors
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