Correlation Between Expressing Of Sirt3?HIF-l? And Activity Of MnSOD In ICC

Objective:To detect the expression of Sirt3 in intraphepatic chol angiocarcinoma tissues and adjacent normal tissues by using Immun ohistochemistry,and to analyze the relationship between Sirt3 expres sion and clinical data and pathological data;The expression of Sirt 3 protein and HIF-1 ? protein was detected by Western blot in hum an cholangiocarcinoma tissues and adjacent normal tissues,WST-1 method was used to detect the MnSOD's enzyme activity of cholan giocarcinoma tissues and adjacent normal tissues.Then the data of Western-blotting result is used to detect the relevance of Sirt3 prote in,HIF-1 a protein a-nd enzyme activity of MnSOD.Method:Collect 33cases of ICC tissues and the adjacent normal tissues which were removed by surgery from January 2015 to October 2016 into the study.Pathological staging of the 33 cases were staged by the 7th TNM staging system of the American Joint Committee On Cancer.The ICC tumor tissue was used as the experimental group and the adjacent normal tissue as the control group.Immunohistochemical method was used to detect the Positive expression of Sirt3 in ICC tissues and adjacent normal tissues,the expression of Sirt3 and HIF-1 a protein in ICC tissues and adjacent normal tissues was quantitatively determined by the Western-blotting method,and the enzyme activity of MnSOD in ICC tissues and adjacent normal tissues was determined by the WST-1 method.Statistically analyzed the data of Immunohistochemical result to determine the correlation between Sirt3 positive expression and clinicopathological data,and to determine the correlation between expression of Sirt3 and HIF-1 a expression and MnSOD activity.Results:1.Immunohistochemical results:The positive expression rate of Sirt3 in ICC tissues was 45.5%(15/33),which is significantly lower than the rate of adjacent normal tissues(100%,33/33),(P<0.05).Sirt3 positive expressing rate is related to lymph node metastasis,differentiation and TNM stages of ICC tumors;that is not related to age?gender?size of tumor,this result is statistically significant(P<0.05).2.Western-blotting results:The relative expression of Sirt3 in ICC tissues is(0.349±0.194),which is significantly lesser than that in the adjacent normal tissues(0.816±0.287),(P<0.05);The relative expression of HIF-1 a in ICC tissues is(0.655±0.105),which is significantly higher than that in the adjacent normal tissues(0.241±0.227)(P<0.05).3.WST-1 results:The activity of MnSOD in the ICC tissues is(0.589±0.094)U/mg,which is significantly lesser than that in the adjacent normal tissues(1.086±0.508)U/mg,with statistically significant(P<0.05).4.Correlation results:The correlation between the expression of Sirt3 and HIF-1 ? is negative,r=-0.911(P<0.01);The correlation between the expression of Sirt3 and the activity of MnSOD is positive,r=0.796(P<0.01);The correlation between the expression of HIF-1 a and the activity of MnSOD is negative,r=-0.741(P<0.01).Conclusion:1.The expressing of Sirt3 is related to lymph node metastasis.The expressing of Sirt3 protein in ICC tissues is lower than adjacent normal tissues,the expressing of HIF-1 a in ICC tissues is higher than adjacent normal tissues,the activity of MnSOD in ICC tissues is lower than adjacent normal tissues.The correlation between the expressing of Sirt3 and the expressing of HIF-1 a is negative,the correlation between the expressing of Sirt3 and the activity of MnSOD in ICC tissues is positive,the correlation between the expressing of HIF-1 a and the activity of MnSOD in ICC tissues is negative.2.For ICC tumor,the low expressing of Sirt3 protein could related to Tumorigenesis,Hypoxic tumor microenvironment and the high level of ROS,it may promote ICC tumor by improving the HIF-1 a-dependent proliferation factors and ROS-related-oxidative damage.