| Objective: We analysis the protein molecular weight in blood serum frompatients of colorectal cancer and colorectal cancer-hepatic metastasis and normalcontrol by Surface enhanced laser desorption ionization-time of flight-massspectrometry (SELDI-TOF-MS),in order to screen relative protein from specificityserum of the patients, and search for specifical Biological Markers of colorectalcancer hepatic metastasis. Meanwhile, by use of computer software(CiphergenProtein Chip),we construct a diagnosis model of colorectal cancer hepatic metastasis.It's very important for the clinical application in valuation of prognostic colorectalcancer hepatic metastasis and early diagnosis of colorectal cancer hepatic metastasis.Methods: 128 serum specimens were collected, including 48 normal control, 44colorectal cancer and 36 colorectal cancer hepatic metastasis. Among the group ofcolorectal cancer, there were 28/44 rectal cancer patients, 16/44 colon carcinomacancer patients. They were all diagnosed by the pathologic assay after operation. Inthe group of colorectal cancer hepatic metastasis all the 36 patients were diagnosed adefinitely through the imageology exam. By the IMAC3 chip of American'sCiphergen company and surface-enhanced laser desorption/ionization time-of-flightmass spectrometry, the protein contents of serum of all the patients and normal control were detected. It was set that the protein molecular weight ranging from 1,500to 20,000 Da for all the detected serum specimens. By the PBS2 protein chip reader,the IMAC3 chip was detected then the protein would be transformed to spectrum. Allthe 3 groups patients were divided into 5 comparison groups and all the 5 comparisongroups are analyzed by the SELDI-TOF-MS. The 5 comparison groups were①.colorectal cancer,colorectal cancer hepatic metastasis.②. colorectal cancer,control.③.colorectaI cancer,colorectal cancer hepatic metastasis and control,.④.Dukes B ofcolorectal cancer, colorectal cancer hepatic metastasis.⑤. Dukes C of colorectalcancer, colorectal cancer hepatic metastasis. By use of "Biomarker Wizard", thedifferent protein content and M/Z in each of the 5 groups were compared. The database was built by using the protein content of these groups which had significantdifference (P＜0.05). And Biomarker Pattern statistic analysis was used to choosecondition accordingly to statistic in group. Therefore the specifical BiologicalMarkers were selected and the diagnostic cast of colorectal cancer hepatic metastasiswas constructed. Also, It was compared the diagnosis model above in colorectalcancer hepatic metastasis with CEA assay. Another group of 27 colorectal cancerhepatic metastasis patients were set, which all be made by definite diagnosis throughthe imageology, and they all experienced the post operation of pathologicdiagnosis,and the group of case had no other relative sickness that influence theprotein content of serum. The Test Data of Biomarker Pattern software was used todetect the same group itself. Meanwhile the CEA serum detection was used ascontrol.Results:.①. Compared colorectal cancer with colorectal cancer hepaticmetastasis: The contents of 25 proteins in the two groups were significantly different(P＜0.05), 9 proteins express lower in colorectal cancer hepatic metastasis. Adiagnosis model was made up of the protein with the relative molecular weight at M5909,M5341,M2685,M2871,M3928,M6635,M8933 respectively.Among them38 of 44 colorectal cancer patient were divided into groups correctly., 36 colorectalcancer hepatic metastasis patient are identified correctly, and the accurate rate is92.5%(74/80). sensitivity 100%(36/36), specificity 86.36%(38/44)②. Comparedcolorectal cancer with control:18 protein content of M/Z showed statisticalsignificance (P＜0.05). 3 of M/Z was choiced as Biological Markers to construct thebest diagnosis model of colorectal cancer hepatic metastasis. The accurate rate is97.8%(90/92), sensitivity 95.5%(42/44), specificity 100%(48/48).③.colorectalcancer,colorectal cancer hepatic metastasis and control: 22 protein content of M/Zshowed statistical significance (P＜0.05). 12 of M/Z was choiced as BiologicalMarkers to construct the best diagnosis model of colorectal cancer hepatic metastasis.The accurate rate is 96.9%(124/128), sensitivity 91.6%(33/36), specificity98.9%(91/92).④. Compared Dukes B of colorectal cancer with colorectal cancerhepatic metastasis:11 protein content of M/Z showed statistical significance(P＜0.05). 11 of M/Z was choiced as Biological Markers to construct the bestdiagnosis model of colorectal cancer hepatic metastasis. The accurate rate is90.32%(56/62), sensitivity 88.89%(32/36), specificity 92.31%(24/26).⑤.Compared Dukes C of colorectal cancer with colorectal cancer hepatic metastasis: 11protein content of M/Z showed statistical significance(P＜0.05).3 of M/Z was choicedas Biological Markers to construct the best diagnosis model of colorectal cancerhepatic metastasis. The accurate rate is 100%(48/48), sensitivity 100%(36/36),specificity 100%(12/12).⑥,The sensitivity and specificity to diagnose patients withcolorectal cancer hepatic metastasis by the way of SELDI-TOF mass spectrometrywere 85.19% (23/27), 70.37% (19/27) separately. The statistics method of fourfoldtable X~2 test was used to analyze the difference between the two patterns (CEA serumdetection/SELDI-TOF mass spectrometry analysis). The result was X~2=1.714, v=1, P=0.190(both side),which indicate no statistically significant difference.Conclusion: This research was to discover the proteome configuration used fordiagnosis of colorectal cancer hepatic metastasis through the analysis of serumprotein. The SELDI-TOF-MS technology was used to analyze the serum proteincontent of normal people, colorectal cancer patient and colorectal cancer hepaticmetastasis patient, and the specifically Biological Markers was screened as earlydiagnosis of colorectal cancer hepatic metastasis in clinic. Compared with traditionaltumor maker CEA detection, this method was easier, operative, faster, more sensitive,and have higher specificity. Biological Markers screened by this method coulddiagnose accurately colorectal cancer hepatic metastasis. Although the two ways indetection patients with colorectal cancer hepatic metastasis did not show statisticallysignificant difference, the lack of samples might be the main reason. It's veryimportant for the clinical application in valuation of prognostic colorectal cancerhepatic metastasis and early diagnosis of colorectal cancer hepatic metastasis. Withthe enlargement of specimen quantity, this technology can be treated as a fasthighly-efficient and sensitive way in early clinical diagnosis of colorectal cancerhepatic metastasis.
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