Significance Of Expression Of CD133and BRAF In Colorectal Cancer

[objective]1.To assessment of the feasibility of microarray technology combined immunohistochemical techniques in the large-scale retrospective trials.2.To explore the CD133protein level by using immunohistochemistry (IHC) technology and the relevance to the clinicopathological characteristics as well as prognosis of colorectal cancer patients.3.To evaluate the BRAF protein and BRAF gene level by immunohistochemistry (IHC) and the relevance to the clinicopathological characteristics as well as prognosis of colorectal cancer patients.[methods]1. Part1:collection2004-2010, fuzhou general hospital of nanjing military region surgical removal of the complete specimens of colorectal cancer diagnosed by pathological and clinical data of400cases, build the organization chip,-using tissue microarray immunohistochemical technique detect colorectal BRAF/CD133protein expression in colorectal cancer tissue;Tissue microarray and conventional pathological comparative research.2. Part2:EliVisionTM plus immunohistochemical staining methods CD133and mutated BRAF protein protein was detected in specimens of400cases of colorectal cancer tissue microarray expression, analyze its correlation with clinical pathological indicators.3. Part3:randomly selected126cases of BRAF immunohistochemical positive and20cases were negative, the mutated BRAF gene mutation situation with direct sequencing method and analyze its relationship with the prognosis of patients with clinical pathological indexes and genes to the clinicopathological characteristics as well as prognosis of colorectal cancer patients.[Results]immunohistochemical results:1. Array wax block complete, no cracking phenomenon, organization wax core order, no shift. The microscope observation does not exist invalid organization, the colored cells for claybank, background without coloring. For CD133and BRAF respectively for regular section and tissue microarray is compared, and found that the coincidence rate reaches as high as93%, analyzes its both Kappa coefficient were0.714,0.700, shows both high coincidence rate, with statistical significance (P>0.05).2. High expression of CD133and tumor differentiation degree and lymph node metastasis had significant correlation (P<0.05), compared with patients age, gender, tumor location, size and differentiation degree, infiltration depth, lymph node metastasis and distant metastasis had no obvious correlation. Kaplan Meier-survival analysis showed that CD133low expression group3year survival rate was96.2%, the high expression of CD133group rates were82.6%at3years, which has obvious difference (P<0.01)3.376cases of colorectal cancer tissue (60.1%) of the BRAF is positive, the mutated BRAF highly expressed positive expression rate of38.8%. With the correlation degree of differentiation in patients with sex, and distant metastasis (P<0.05); But with age, tumor location, size, infiltration depth, lymph node metastasis has no correlation (P>0.05); BRAF survival rate low negative positive patients, but there was no significant difference statistically, its expression level also and survival in patients with no significant correlation (P>0.05); By Spearman correlation analysis was carried out on the protein CD133and mutated BRAF protein, is found that both have certain correlation (r=0.505)gene mutation test results:Total mutated BRAF gene mutation in colorectal cancer was8.9%(13/146), the mutation was associated with a significant staging and survival in patients with colon cancer, with statistical significance (P<0.05).There was associated relationship between BRAF gene mutation and survival time.[Conclusions]1. The TMA technique provides a reliable method to investigate marker expression by offering a rapid, economic and accurate screening of tissue specimens on a large scale.2. CD133expression in colorectal cancer was significantly higher, and is closely related to the low tumor differentiation, lymph node metastasis; CD133and can be used as judgement index of prognosis of patients with colorectal cancer, the expression level is higher, the worse prognosis.3.Though BRAF strongly positive and gene amplification are concordance in some degree, IHC high expression cases should be further detected by direct sequencing to assessBRAF gene status in colorectal cancer. And patients with BRAF amplification also might constitute potential candidates for targeted therapy with humanized monoclonal antibodies.