| Canine neoplastic disease is a common disease.With the improvement of people’s living standards and the development of veterinary industry,more and more attention has been paid to canine cancer.The main types of tumor in dogs are breast tumor,skin and soft tissue tumor,perianal tumor,testicular tumor and vaginal tumor.Breast tumor is one of the most common tumors in clinic,which is more likely to occur as the bitch ages longer.To investigate the incidence of common tumors in canines,271 cases of dog tumors from some pet hospitals in Harbin were collected from 2014 to 2017.The basic information of each clinical tumor case was collected including,species,gender,age,tumor sites and diet,and the data were statically analyzed.In order to accurately determine the benign and malignant of these 271 canine cancer cases,histopathologyic examination were performed.Enzyme-linked immunosorbent assay(ELISA)were conducted to screen 12 serum tumor markers of CA15-3,CEA,SF,VEGF,CA199,CA125,SCCAg,NSE,HER-2,CYFRA21-1,CYFRA21-1 and HER-2 in 138 cases of breast tumor,aiming to find four effective early diagnostic serum tumor markers with high sensitivity and specificity for canine mammary gland tumor in joint detection.Accordingly,the expression of mRNA and protein of these four markers were also investigated in canine mammary tumor tissues using real-time fluorescent quantitative PCR(qRT-PCR)and Western blot.Results :(1)Among these 271 confirmed tumor cases,80 were malignant tumors and accounted for 29.5% of all tumors.In all the cases,mammary glands and skin were high-risk sites for canine tumors,accounting for 50.9% and 31.4% respectively.138 breast tumors cases were determined,in which 56 cases were malignant,accounting for 40.6%.Dogs’ age ranged from 1 to 17 years old.The average age was 9.6 years,while the tumor most occur in age 7 and older.Old and middle-aged dogs were still the main group,but the incidence of young dogs is increasing year by year.All varieties of dogs can have tumors,but mixed dogs had the highest incidence,followed by the poodle and Pekingese,and the type of tumor was mainly breast cancer.Diet is mainly leftovers and meat rice type accounted for 42.4% and 32.5% of the total number of dogs,respectively.Some tumor types were related to the dog’s sex and whether it was neuter.(2)The ELISA method was used to detect the tumor markers in the serum,and the difference among the breast cancer group with the benign breast disease group and the healthy control group were statistically significant(P<0.05).There was no significant difference in other markers between the benign breast disease group and the healthy control group(P>0.05)except that the three markers of SF,VEGF and CA199(P<0.05).(3)In all 12 kinds of tumor markers,4 markers with better sensitivity and specificity were CA15-3,CEA,SF and HER-2.(4)The sensitivity,specificity and accuracy of single test of CA15-3 were 51.8%,93.9% and 76.8%,respectively.The sensitivity,specificity and accuracy of CEA were 44.6%,84.1% and 68.1%,respectively.The sensitivity,specificity and accuracy of SF were 62.5%,85.4% and 76.1%,respectively.The sensitivity,specificity and accuracy of HER-2 were 46.4%,84.1% and 68.8%,respectively.The single detection of SF has the highest sensitivity and while the CA15-3 has the highest specificity.The sensitivity,specificity and accuracy of combined detection of 4 tumor markers were 85.7%,78.1% and 81.2%,respectively.Sensitivity and accuracy increased but the specificity decreased.(5)The relative mRNA expression of CA15-3,CEA,SF,NSE and HER-2 in breast tumor tissues of dogs was detected by qRT-PCR,and the malignant tumor tissues were significantly higher than those of benign breast tumors and normal mammary tissues(P<0.001).Besides the SF gene,there was no significant difference between the benign breast tumor and normal breast tissue(P>0.05)in other markers,which was similar to the result in serum.(6)The protein expression of CEA,SF,NSE and HER-2 in canine mammary gland tumors were detected by Western blot.The expression in malignant tumor tissues were significantly higher than benign breast tumors and normal mammary tissues(P<0.001).Except for SF protein,there was no significant difference between benign breast tumors and normal breast tissues in other marker(P>0.05).Conclusion:This study provides a reference for the epidemiology and diagnosis of canine tumors,and has reference value for the prevention and diagnosis of canine tumors.We screened four effective tumor markers CA15-3,CEA,SF and HER-2 for early diagnosis of dog breast tumors,in which the sensitivity and accuracy of joint detection of these four markers are higher than those of single detection. |
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