The Value Of CHEK2 Gene Mutates And Assay Of TSGF, CA15-3, CA125 And CEA In The Diagnosis Of Breast Cancer.

Objective The cell cycle checkpoint kinase gene (CHEK2 gene) is known as a kind of anti-oncogene. This study was to explore CHEK2 gene mutation in human breast cancer, and assess the value of assay of Tumor Specific Growth Factor (TSGF), CA15-3,CA125 and CEA in the diagnosis of breast cancer. Methods (1). CHEK2 gene: Breast cancer tissues of 65 patients were collected as experiment group, and breast tissues of 21 cases of patients with benign breast disease and blood samples of 145 cases of health persons were randomly collected as control group. DNA of each sample was extracted by the phenol-chloroform method. To determine the concentration and purity of the DNA solution, the absorbance of ultraviolet light was measured in a spectrophotometer. Fragments of CHEK2 gene exon 1,10,11,13,14 were amplified by polymerase chain reaction(PCR). Mutations was studied by single-strand confirmation polymorphism analysis (SSCP), and the alteration were conformed by DNA sequencing. (2) Serum TSGF, CA15-3,CA125 and CEA: 69 patients with breast cancer were studied as experiment group, and 31 cases of patients with benign breast disease were randomly selected as control group. 3ml fast blood sample was taken from each patient, and serum was separated by centrifugation. Serum level of TSGF in each patient was determined with chromatometry method, and serum levels of CA15-3,CA125 and CEA were determined with chemoluminescence method. Statistical analysis was performed by using SPSS. Results (1),CHEK2 gene: 4 single nucleotide changes in breast cancer patients were identified, including 1 frame-shift mutation in exon 10(1100 del C) and 3 insertion mutations in exon 11(1336 ins G) of CHEK2 gene. In addition, we found 1 missense mutation in exon 1(252A>G) of CHEK2 in health persons. Mutations in exon13 and exon14 were not identified in our study. The mutational rate of CHEK2 gene was 6.15%(4/65)in breast cancer patients, and was 0.6%(1/166) in control group, and the difference between the two groups was significant (P<0.05). (2),The serum concentration of TSGF,CA15-3 and CEA in breast cancer group was higher than that in benign breast disease group, P<0.05.There was no difference in CA125 between the two groups, P>0.05.The positive rates of TSGF, CA15-3,CA125 and CEA in breast cancer were 21.7%,30.4%,13.0% and 15.9%,respectively,and the specificity rates were 100%,93.5%,83.8%,100%, and the accuracy rates were 46%,50%,35% and 42%,respectively.In combined detection of TSGF,CA15-3 and CEA, The positive rate was 62.3%, the specificity rate was 93.5%, and the accuracy rate was 72.0%。comparing with single assay, combined analysis of TSGF,CA153 and CEA has a higher diagnostic sensitivity. Conclusions Mutation in CHEK2 is closely related to breast cancer, and CHEK2 gene mutation may play an important role in the development of sporadic and hereditary breast cancer in human. The assay of TSGF, CA153 and CEA was significant in the diagnosis of breast cancer, combined analysis of TSGF, CA15-3 and CEA can obviously improve the diagnosis rate.