Expression And Significance Of CYP1B1 In Human Breast Fibroadenomas

ObjectiveTo investigate the expression of cytochrome P450(CYP1B1)in human breast fibroadenomas and its relationship with clinicopathological parameters,and to explore its possible mechanism to provide a theoretical reference for the prevention and treatment of breast fibroadenomas.MethodsFrom November 2016 to March 2017,80 patients with breast fibroadenomas were treated with breast surgery at the Fifth Affiliated Hospital of Zhengzhou University.The patients were all female.The preoperative imaging data was complete.All specimens were 10% formaldehyde.Fixed,paraffin embedded.The patients were 16 to 50 years old,with an average age of 31 years and a median age of 30 years.38 cases were ?29 years old,42 cases were >29 years old.No radiotherapy,chemotherapy or other medications were given before surgery.All patients have complete medical records.All matters in this study have been informed and agreed by the patient's family and hospital ethics committee.All specimens were fixed in 10% formaldehyde after excision,routinely paraffin-embedded,etc.,and the embedded specimens were serially sectioned at 4 ?m thick and placed on a slide after pretreatment with anti-offset agent.Add CYP1B1 antibody(diluted 1:200)to incubate overnight at 4 °C in a refrigerator;wash with PBS buffer,add reaction enhancer,enhance the enzyme-labeled goat anti-mouse/rabbit IgG complex,and show DAB color Color observation;hematoxylin counterstaining,clear water back to the blue after the neutral gum seal,observed under the microscope.The primary antibody was replaced with PBS buffer as a control.Two pathologists used double-blind method for reading and scoring.Eight high-power fields were randomly observed under light microscope,and CYP1B1 positive cells were counted.Among them: the percentage of positive cells ?25% is 1 point,> 25% to 50% is 2 points,> 50% ~ 75% is 3 points,> 75% is 4 points;according to the degree of staining: 0 points without dyeing The light yellow color is 1 point,the yellow or brown color is 2 points,and the brown one is 3 points.Finally,the sum of the two scores is the final score: 0 is negative,1 to 7 is positive,1 to 3 is low,and 4 to 7 is high.Color Doppler score: routine mode ultrasound to find the lesion,select the ultrasound elastography mode,obtain the imaging image,and use Luo Baoming's improved 5-point method to score.The data was processed and analyzed using the biometric software IBM SPSS 25.0.The KS method was used to test the normal distribution of the data.The quantitative data were expressed as mean±standard deviation(x±s).Quantitative data between the two groups were tested by t test;the count data was calculated by chi-square or fisher exact probability test;P<0.05 was considered statistically significant.ResultCYP1B1 was highly expressed in the cytoplasm of tumor tissues,and yellow or brownish yellow particulate matter could be seen.The dark staining was brown,as shown in Fig.1.There was a statistically significant difference in the expression of CYP1B1 in tumor tissues and normal peritumoral tissues(?2=36.304,P<0.05).The CYP1B1 strip optical density value measured for each specimen was compared with the internal reference GAPDH strip optical density value,and the ratio obtained was the CYP1B1 protein expression level in the specimen,as shown in Fig.9.The negative expression of CYP1B1 in normal breast tissue was 0.10±0.05,and that in tumor tissue was 1.56±0.13.The data were normalized by K-S method,z=0.495,P=0.967.The expression of CYP1B1 in tumor tissues was significantly higher than that in normal breast tissue,and the difference was statistically significant(t=21.045,P<0.005).The expression of CYP1B1 was correlated with ultrasound UE score and tumor diameter(P<0.05),and was not related to patient age and tumor location(P>0.05).Conclution1.CYP1B1 is highly expressed in human breast fibroadenoma tissue,with little or no expression in normal breast tissue.2.The expression level of CYP1B1 is not related to the location of the tumor and the age of the patient.It is associated with the diameter of the adenoma and the UE score of the breast,and the expression of CYP1B1 is higher in the larger tumor diameter and higher breast color UE score.