: 10389822 : 1
: TNM TNM 2
p27 kip1 MMP-7 MMP-9 Cyclin E TGF-β1 p53 114 1999 2004 SCI >1.4 6 p27 kip1 MMP-7 MMP-9 Cyclin E TGF-β1 p53 1995-1999 114 114 Kaplan-Meier Log-rank 6 p27 kip1 MMP-7 PearsonX 2 p27 kip1 MMP-7 COX p27 kip1 MMP-7 PM PM TNM PM 16 3
p27 kip1 Western blotting 25 p27 kip1 p27 kip1 mrna RT PCR 16 p27 kip1 mrna p27 kip1 p27 kip1 p27 kip1 : 1. p27 kip1 MMP-7 114 p27 kip1 p27 kip1 88.40 vs 52.94 P<0.01 MMP-7 MMP-7 89.27% vs 70.59% P<0.05 Cyclin E MMP-9 TGF-β1 p53 2. p27 kip1 MMP-7 p27 kip1 TNM MMP-7 3. COX MMP-7 p27 kip1 4. p27 kip1 MMP-7 PM A B C COX PM p27 kip1 MMP-7 5. PM 114 4
PM 7.69 PM 21.43% PM 71.43 P<0.05 6. PM TNM PM PM 7. P PM P-PM 114 P-PM 8. p27 kip1 9. RT-PCR p27 kip1 mrna p27 kip1 mrna 93.75 p27 kip1 p27 kip1 RT-PCR : 1. p27 kip1 MMP-7 2. p27 kip1 TNM MMP-7 TNM P 0.076 3. PM p27 kip1 MMP-7 4. PM 5
5. P PM P-PM P-PM PM 6. 7. p27 kip1 mrna p27 kip1 RT-PCR : p27 kip1 MMP-7 6
The relevance of biomarkers panel in stage colorectal cancer and M.D candidate: Supervisor: Leng Jiahua Prof. Gu Jin Prof. Li Jiyou Abstract Background: Colorectal carcinoma is one of the most common tumors which have poor prognosis. Although pathologic stage is currently the main prognostic indicators for patients with CRC, mounting evidence suggests that, in the current form, it is insufficient to predict clinical outcome. Patients with same TNM stage and same differentiation would have markedly different prognosis. This condition directly make it difficult for doctors to plan individual adjuvant therapies for each postoperative patient. Thus over-treatment and insufficient-treatment become inevitable. Examination with biomarkers panel may be possible to identify aggressive tumor phenotypes, improve clinical staging and provide useful information for the application of individual therapeutic strategies. Objective: The objective of the study is to investigate the prognostic significance of multiple biomarkers in stage and CRC. Screen out an optimal biomarkers panel, and evaluate it s prognostic significance in different subgroups of stage and CRC. Further, try to build up a new prognostic classification system which combined with clinical pathological factors and biomarkers panel. It might lead to a more reliable prognostic estimation. Methods: In this study we retrospectively analyze The expression of p27 kip1 MMP-7 MMP-9 Cyclin E TGF-β1 p53 by immunohistochemistry in 7
paraffin embedded specimens from 114 consecutive patients with stage and CRC who underwent resection for colorectal carcinoma between 1995 and 1999 in our hospital. Multivariate analysis with COX model is applied to indicate the independent prognostic biomarkers of colorectal cancer. The set of biomarkers are studied in each stage of colorectal carcinoma. Analyze the feasibility on building up a colorectal carcinoma prognostic classification system combined with clinical pathological factors and biomarkers panel. Repeated immunohistochemistry investigation and Western blotting examination were applied to confirm the repeatability and reliability of the IHC result. Reverse transcription polymerase chain reaction were also done in 16 fresh tissue sample to detect their mrna and protein expression in contrast to IHC. Results: 1. Expression of p27 kip1 and MMP-7 are significantly associated with prognosis of stage and CRC in univariate analysis. Over-expression of p27 kip1 is a protective factor for patient s outcome, on the contrary over-expression of MMP-7 is a risk factor. P 0.01, P 0.05 respectively. 2. Expression of p27 kip1 is associated with TNM stage and histologic grade. Expression of MMP-7 is correlated with the gross configuration of tumor. 3. Multivariate analysis indicated that p27 kip1, MMP-7 and histologic grade were independent predictors of prognosis in stage and CRC. P 0.01, P 0.05, P 0.05 respectively. 4. The coexpression panel of p27 kip1 and MMP-7 have significant prognostic value in stage and CRC patients. Patients with two protective factors in PM panel had a good-prognosis, while patient with two risk faors in PM panel had a poor-prognosis, p 0.01, the mean (±SE) 5-year disease free survival was 92.26±3.33 and 28.57± 17.07percent respectively. 5. Multivariate analysis indicated that The biomarkers panel PM(p27 kip1, MMP-7) was also independent prognosis predictor in stage and CRC. P 0.001. 6. If PM have two risk factors, then the possibility of developing recurrece and metastasis was 71.43%. While, when PM have two protective factors the risk was only 7.69%. 7. PM could show it s prognostic value even in the presence of traditional clinical and pathological criteria such as histologic grade and TNM stage in stage and CRC. 8. This result also indicates that both sets of the prognostic criteria misclassify a significant number of 8
patients. While the new system which was combined with clinical pathological factors and biomarkers panel lead to a more reliable prognostic estimation. 9. p27 kip1 mrna have a stable expression. Positive expression was observed in 15 out of 16 stage and CRC tumor sample. 10. Useing IHC to evaluate biomarker s expression level have markedly repeatability and reliability. Conclusions 1. Based on the evidence of this study, p27 kip1 and MMP-7 seem suitable candidates for a selective panel of biomarkers designed to provide significant additional information with respect to the current clinical pathological staging system in stage and colorectal cancer. 2. The PM panel is a more powerful predictor of the outcome of disease in stage and CRC than traditional systems based on clinical and pathological criteria. 3.Combined clinical pathological factors with biomarkers panel PM could provide prognosis estimation in a more accurate way, and thus might be helpful for further individualized treatment. Key words: biomarkers panel, stage and colorectal cancer, p27 kip1, MMP-7, prognosis. 9
Biomarker p27 kip1 MMP-7 MMP-9 Cyclin E TGF-β1 CDK CKI ECM TIMP CRC IHC PM -7-9 E β1 p27 kip1 + MMP-7 10
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1,023,000 529,000 (1) 2005 145,290 56,290 (2) 80 10/10 90 21-22/10 (3) (4) 92 84 (5) 50 (6) 20 70 (7) 5 45 (8) (7) (9) (10) 40 (11) 60 12
TNM (5) TNM TNM TNM (12) TNM (13) (14-16) 13
6 114 CRC 6 114 CRC Kaplan-Meier COX p27 kip1 MMP-7 CRC PM p27 kip1 MMP-7 RT PCR mrna P PM P-PM CRC 14
SCI 1999 2004 impact factor >1.4 80 TNM AJCC (17) 15
p27 kip1 MMP-7 MMP-9 Cyclin E TGF- 1 p53 p27 kip1 MMP-7 CRC PM p27 kip1 MMP-7 CRC PM PM PM PM P PM P-PM 16