Diagnostic And Differential Value Of Elevated Serum And Bile MMP-7Levels In Cholangiocarcinoma

Objective1.To determine the serum MMP-7levels in patients and investigate theirdiagnostic value.2.To determine the bile MMP-7levels in patients and investigate their diagnosticvalue.Materials and Methods1.Collected from December2011to June2012in the Eastern HepatobiliarySurgery Hospital Endoscopy Department29patients with cholangiocarcinoma(CCA)and20benign patients serum and bile samples.2.MMP-7and clinical parameters testing(1). Extraction of fasting venous blood in patients with3ml, allow samples toclot for3-4hours at room temperature. Centrifuge at approximately3000rpm for15minutes.Remove serum and store at-80℃.(2). Before the injection of contrast agent collect bile and above all is due toobstruction of bile or narrow segment. Centrifuge at approximately3000rpm for15minutes.Remove the supernate and store at-80℃.(3).ELISA was used to measure the serum and bile level of MMP-7. Secure thedesired numbers of coated wells in the holder then add100μL of standards or theappropriate well in the antibody pre-coated Microtitier Plate. Add100μL of PBS inthe blank control well. Add50μL of conjugate to each well. Mix well. Cover andincubate the plate for1hour at37℃. Wash plate five times with diluted wash solutinusing auto washer.After washing, dry the plate. It is recommended that the washer beset for a soaking time of10seconds and shaking time of5seconds between eachwash. Add50μL substrate A and substrate B to each well including blank controlwell,subsequently. Cover and incubate for10-15minutes at20-25℃. Add50μL ofstop solution to each well including blank control well.(4).The sensitivity and specificity of MMP-7for the diagnosis of CCA weredetermined, and the relationship between its levels and clinical parameters wereanalyzed.Results 1.The serum MMP-7level in CCA group (M5.47ng/ml,IQR4.26-6.81; x±SD5.42±0.30ng/ml) was significantly higher than benign group (M4.22ng/ml,IQR3.82-4.82; x±SD4.28±0.19ng/ml, P=0.009).Using4.88ng/ml(AUC0.722，95%CI0.580-0.865) as the cutoff point, the sensitivity and specificity of MMP-7fordiagnosis of CCA was65.5%and80.0%.2.The bile MMP-7level in CCA group (M9.01ng/ml,IQR8.24-10.78; x±SD10.05±0.69ng/ml) was significantly higher than benign group (M7.64ng/ml,IQR7.02-8.70; x±SD8.26±0.443ng/ml, P=0.012). Using8.32ng/ml(（AUC0.713，95%CI0.564-0.862) as the cutoff point, the sensitivity and specificity of MMP-7fordiagnosis of CCA was75.9%and65.0%.3.The serum and bile level of MMP-7was not associated with clinicalparameters.ConclusionThe serum and bile MMP-7might be useful markers of CCA，and it play a rolein the differential diagnosis between CCA and benign disease.