| Part One Clinical significance of peripheral blood micro-metastasis in colorectal cancerObjective Peripheral blood micro-metastasis is one of the important factors for failure of surgical resection in colorectal cancer.Above 50%of the patients underwent surgical resection died of cancerous relapse or metastasis,indicating that micro-metastasis may have exited before the sugeries.CK20 mRNA has thought to be a specific biomarker for early diagnosis of micro-metastasis in colorectal cancer.To explore the role of determination of peripheral blood micro-metastasis in prognosis of colorectal cancer and to explore whether microvessel density(MVD) and lymphatic microvessel density(LMVD) have correlation with peripheral blood micro-metastasis.Methods Using CK20 mRNA as a specific biomarker of tumor cells in peripheral blood,we determined its levels before and after surgical resection with the RT-PCR technique,also we determined the MVD and LMVD levels by immunohistochemistry.Results Eighteen of the 51 patients were found positive CK20 mRNA expressions before surgical resection,of them,1(12.50%) was of the 8 patients with stageâ… ,4 (23.53%) was of the 17 patients with stageâ…¡,6(35.29%) was of the 17 patients with stageâ…¢,and 7(77.78%)was of the 9 patients with stageâ…£.These findings indicated that CK20 mRNA expression in peripheral blood correlated with the TNM classification in colorectal cancer.After surgical resection,3 more patients with rectum cancer were also shown positive CK20 mRNA expressions.CK20 mRNA expression had no relations to age,gender,primary location,tumor size,peripheral blood carcino-embryonic antigen level,and tumor differentiation and invasion,but had relations to lymphonode metastasis,distant metastasis and tumor TNM stage (P＜0.05).The MVD levels had no relations to age,gender,primary location,tumor size and the peripheral blood carcino-embryonic antigen level,but had relations to tumor differentiation and invasion,lymphonode metastasis,distant metastasis,tumor TNM stage(P＜0.05).Also the LMVD levels had no relations to age,gender,primary location,tumor size,the peripheral blood carcino-embryonic antigen level,and tumor differentiation and invasion,but had relations to lymphonode metastasis,distant metastasis,and tumor TNM stage(P＜0.05).The MVD and LMVD levels were 58.83±18.28 and 9.72±1.64 in the positive CK20 mRNA group,however they were 30.48±9.31 and 8.21±1.87 in the negative CK20 mRNA group,the difference was statistically significant(P＜0.01 ).Conclusions Peripheral blood micro-metastasis is related to lymphnode metastasis, distant metastatis and TNM stage.Surgical procedures may promote the genesis of periphral blood micro-metastasis.Determination of tissue vascularization and genesis of lymphatic vessel is helpful in judging genesis of peripheral blood micro-metastasis in colorectal cancer.Part Two Tiam1 and DcR3 expressions in colorectal cancer and their correlation with peripheral blood micro-metastasisObjective Invasion and metastasis are key characteristics of malignant tumors.It has been shown that T lymphoma invasion and metastasis factor 1(Tiam1) closely correlates with tumor invasion and metastasis.Decoy receptor 3(DcR3) is one member of the tumor necrosin receptor hyperfamily,and it is overexpressed in many human malignant tumors,also its overexpression closely relates to tumor lymphnode metastasis and pathologic staging.To determine the expressions of Tiam1 mRNA and DcR3 mRNA in cancerous tissues and non-cancerous tissues of colorectal cancer,and to explore their correlation with pathologic data and peripheral blood micro-metastasis.Methods Using the RT-PCR method,the CK20 mRNA expression was determined.Using fluorescence quantitative-RT-PCR(FQ-RT-PCR),the expressions of Tiam1 mRNA and DcR3 mRNA were determined in 51 cases of colorectal carcinoma.Results The Tiam1 mRNA expression level was(20.61±4.99)×10-4 in cancerous tissues and was(3.49±2.74)×10-4 in non-cancerous tissues(P＜0.01).Also it was (72.11±2.38)×10-4 in cancerous tissues with micro-metastasis and was(15.78±5.01)×10-4 in cancerous tissues without micro-metastasis,the difference was statistically significant(P＜0.05).Tiam1 mRNA expression had no relations to age,gender, primary location,tumor size and the peripheral blood carcino-embryonic antigen level,but had relations to tumor differentiation and invasion,lymphnode metastasis, distant metastasis and tumor TNM stage(P＜0.05).The DcR3 mRNA expression level was(42.40±2.83)×10-4 in cancerous tissues and was(12.59±2.55)×10-4 in non-cancerous tissues(P＜0.01).Also it was(61.53±2.60)×10-4 in cancerous tissues with micro-metastasis and was(34.60±2.84)×10-4 in cancerous tissues without micro-metastasis,but the difference was not statistically significant(P＞0.05).DcR3 mRNA expression had no relations to age,gender,primary location,tumor size and peripheral blood carcino-embryonic antigen level,but had relations to tumor differentiation and invasion,lymphnode metastasis,distant metastasis and tumor TNM stage(P＜0.05).Conclusions Overexpressions of Tiam1 mRNA and DcR3 mRNA correlate with genesis of colorectal cancer.Tiam1 mRNA overexpression is a high risk factor for peripheral blood micro-metastasis in colorectal cancer,but DcR3 mRNA can not be used as a high risk factor for peripheral blood micro-metastasis in colorectal cancer. Part Three Human KaUikrein6 expression in colorectal cancer and its correlation with peripheral blood micro-metastasisObjective Kallikreins(KLKs) have a similar genomic organization and show significant homology at both the nucleotide and the protein level.Human KLK gene family has participated in genesis and development of cancers.A growing body of evidence suggests that KLKs are involved in human malignancies and that many KLKs are promising biomarkers of prostate,ovarian,breast and testicular cancers.To determine the expression of KLK6 mRNA in colorectal cancer and to explore its relationship to clinicopathologic features and peripheral blood micro-metastasis in colorectal cancer.Methods Using the RT-PCR method,the CK20 mRNA expression was determined.Using the FQ-RT-PCR method,the KLK6 mRNA expression was determined in 51 cases of colorectal carcinoma.Results The KLK6 mRNA expression level was(33.11±2.34)×10-4 in cancerous tissues and was(12.88±2.51)×10-4 in non-cancerous tissues(P＜0.01).Also it was (40.64±2.05)×10-4 in cancerous tissues with micro-metastasis and was(30.06±2.45)×10-4 in cancerous tissues without micro-metastasis,the difference was not statistically significant(P＞0.05).KLK6 mRNA expression had no relations to age, gender,primary location,tumor size,peripheral blood carcino-embryonic antigen level,tumor differentiation and invasion,and distant metastasis,but had relations to lymphnode metastasis and tumor TNM stage(P＜0.05).Conclusions Overexpression of KLK6 mRNA correlates with genesis of colorectal cancer,however,it can not be used as a high risk factor of peripheral blood micro-metastasis in colorectal cancer.
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