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Objective: To analyze the mutations of BRCA1 in 9 Chinese familiar breast cancer patients. Methods: Peripheral blood samples were obtained from 9 patients enrolled from 9 breast cancer families, one normal control, 32 sporadic breast cancer patients and 33 normal donors. DNA extracted from lymphocytes was amplified by polymerase chain reaction (PCR). The 22 exons and partial introns of BRCA1 were screened by PCR denaturing high performance liquid chromatography (PCR-DHPLC) and confirmed by direct sequencing. Results: Among these 9 familiar breast cancer patients, a deleterious mutation was detected in one case in exon 11 (3870delTGTC) which was a 4 base deletion and caused a frameshift in turn. One novel and unique amino acid substitution (E867R) was detected in one case. Eight patients were detected to have a known variation in intron 18 (IVS18+65G→A), and the ratio of this variation detected was 88.9%(8/9). The ratio of this variation was 37.5%(12/32) in sporadic breast cancer patients or 33.3%(11/33) in normal control. This variation was found to be accompanied all the time with a known missense variation in exon 11 (P871L) and a polymorphism in intron 9 (IVS8 57delT). Those three variants were also detected in homozygous in one case, which implies the linkage of the 3 sites. The linkage had not been reported. Two patients had been found with a known polymorphism in exon 13 (S1436S). Another known polymorphism was found in one case (L771L). In addition, intronic variants (IVS2+48C→T, IVS2+133C→T, IVS12+112C→A) were detected. Conclusion: The mutations of BRCA1 in Chinese familiar breast cancer patients are different from the hot spots reported in Caucasian and Jewish. It is important that further study be conducted to seek for specific mutations of this gene or other possible relevant genes in Chinese familiar breast cancer patients.
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Objective To investigate the relationship of c erbB2 with endocrine therapy and prognosis, to investigate the rapid and effective method to detect c erbB2 alteration in breast carcinoma(BC).Metheds Semi quantitative PCR was used to detect the amplification of c erbB2, and immunohistochemistry was used to detect the expression of protein of c erbB2. 58 cases of BC were followed up .Results There was significant relationship between c erbB2 protein overexpression and gene amplification(P
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Objective To explore the role of Bcl-2 and Caspase-3 in modulating apoptosis of ER-negative breast cancer cells induced by tamoxifen. Methods ER-negative breast cancer cell lines MDA-MB-231 were treated with 10.0?M tamoxifen for 12, 24, 36,48, 60 hours. The rate of cell apoptosis with or without caspase-3 inhibitor Ac-DEVD-CHO, and protein expression of Bcl-2,Bax were determined by flow cytometry, and the activity of Caspase-3 was examined with fluorophotometry. Results The expression of Bcl-2 was down-regulated, the activity of Caspase-3 and the rate of cell apoptosis were increased by TAM time-dependently, and the rate of apoptosis reached its peak at 48 hours. The expression of Bcl-2 was (negatively) correlated with activity of caspase-3. Tamoxifen, however, did not affect Bax protein expression. Ac-DEVD-CHO, a caspase-3 inhibitor, blocked the activation of caspase-3 and inhibited cell apoptosis (induced) by tamoxifen. Conclusions TAM could induce apoptosis in ER-negative breast cancer cells via (mitochondria) pathway by down-regulating Bcl-2 expression, and the activation of Caspase-3 might play an (important) role in the process of tamoxifen-induced apoptosis of ER-negative breast cancer cells.
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Objective To clone the transcriptional regulatory sequences (TRS) in human breast cancer related DF3 antigen, and to test the relationship between the activity of the TRS and the cell surface DF3 antigen. Methods Authors designed a pair of primers according to the registered 5′-flanking region of DF3 antigen. The 771 base pairs of DNA fragment were amplified from the genomic DNA of human MCF-7 breast carcinoma cells by PCR, and cloned to the pMD18-T vector. The results were tested by restrictive enzyme analysis and DNA sequencing. The DF3 TRS was cut by double enzyme: Mlu I、Hind III,and cloned to the pGL3 vector . The activity of the DF3 TRS was expressed by analyzing the relative luciferase activities. Results Restrictive endonuclease identification and DNA sequencing proved that the sequence authors got, was correct. The luciferase activity in MDA-MB-231 was hardly detected, whereas in MCF-7 the luciferase activity was about 200 times than in MDA-MB-231. Conclusions The DF3 TRS was cloned successfully. The DF3 activity has a distinct relationship with DF3 antigen. The study shows that DF3 TRS can be used in the gene therapy of breast carcinoma.
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0.05). The expression of VEGF121 mRNA and VEGF165mRNA in BC with lymph node metastasis was significantly higher than that in BC without lymph node metastasis(P=0.033,0.004).Conclusions The expression of VEGF121 mRNA and VEGF165mRNA in BC tissues is upregulated, and VEGF121mRNA is likely to play the major role. A high level expression of VEGF mRNA in BC patient indicates the likelihood of lymph node metastasis.
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Objective To evaluate the relationship between the clinicopathological factors, prognosis and the expression of p16 and c erbB 2 protein in primary breast cancer. Methods The expression of p16 and c erbB 2 by immunohistochemical method was observed in 50 patients with primary breast cancer and the detection of p16 by polymerase chain reaction(PCR) and the point mutation of p16 by PCR single strand conformational polymorphism(SSCP) were detected in 20 patients with breast cancer. Results Among the cancers, positive expression of p16 protein was found in 17(34.00%) cases, c erbB 2 protein positive expression in 24(48.00%) cases. No homozygous deletion in p16 gene was found. However, exon2 point mutation of p16 gene was found in 1 of 20 breast cancer. The results showed no relationship between p16 expression and clinicopathological factor or prognosis. Positive expressions of c erbB 2 protein were often found in breast cancer with lymph node metastasis(P=0.0237) with a poor 5 year survivalrate(P=0.0169). There was no consistent relationship between the expression of p16 and c erbB 2 protein. Neither p16 nor c erbB 2 protein expression could be as an independent prognostic factor. Conclusions The patients with breast cancer of positive expression of c erbB 2 protein has a high lymph node metastasis rate and a poor survival rate. The point mutation rate of p16 gene is lower in primary breast cancer, and it can be a molecular events in advanced primary breast cancer.
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Objective To explore the relationship of bone marrow micrometastases(BMM) with nm23 expression of breast cancer(BC) in patients with stage Ⅰ BC. Methods BMM and nm23 expression of carcinoma tissue in 52 cases of stage Ⅰ BC were examined by immunohistochemical technique with monoclonal anti epithelial membrane antigen(anti EMA) and nm23 H1. Results BMM was observed in 10 of 52 patients(19.2%). In the group of poor differentiated cancer, the positive rate of BMM was significantly higher than that in well differentiated cancer(P
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Objective To explore the expression and biological significance of resistance gene of GST Pi,ToPoII and PgP in breast carcinoma. Method To test and analyse the resistance gene for GST Pi,ToPoII and PgP of 52 cases of breast carcinoma was made by using instant microwave heating immunohistochemistry.Results (1) Positive expression rates of GST Pi,ToPoII and PgP in 52 cases of breast carcinoma were 50.0% (26/52), 17.3% (9/52) and 11.5%(6/52),respectively. (2) Invasive ductal carcinoma had positive expression of three resistance genes at different levels,and the resistance genes in invasive ductal carcinoma were more widely than those in other types breast cancer. (3) There was no obvious relationship between positive expression of GST Pi,ToPoII and PgP in breast carcinoma with patients' age. However,the positive expression of three resistance genes in patients aged 40 to 49 and aged 60 to 69 were higher than other ages.Conclusions (1) The multidrug resistance genes existing in some patients of breast carcinoma or the primary cancer cell has resistance to GST Pi,ToPoII and PgP antineoplastics. (2) Before chemotherapy to patients with breast carcinoma, the test of resistance gene should be done, which will help doctors select a resonable method of combination of drugs so as to improve curative effect and to prolong patients' life.
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Objective To study the expression of fascin, an actin bundling protein associated with cell motility, and the relationship with ER negative breast carcinoma.Methods Comparasion of the expression of fascin, estrogen receptor (ER) and proliferative cell nuclear antigen (PCNA) determined by immunohistological method was made on 84 specimens of primary breast carcinomas. Results The diffuse staining of fascin in cancerous cell cytoplasm was noted. Only 3 of 53 breast carcinomas with ER positive showed fascin positive expression (5.67%); in contrast, 21 of 31 carcinomas with ER-negative showed positive expression(67.7 %) (P
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Objective To assess the clinical significance of the expression of p53, p21 protein in breast cancer (BC). Method The expression levels of p53, p21 proteins were examined by immunohistochemical (IHC) SP techniques in 69 BC specimen and 20 paracancinoma breast tissue. Result p53 and p21 proteins were failed to be observed in the paracancer tissue. p53 protein was found in 47.8% of BC, while p21 protein in 43.5% of BC. Along with the declined of BC cell differentiation degree, p53 expression increased significantly and p21 expression declined obviously. Expression of p21 was lower in patients with lymph node metastasis than that in patients without lymph node metastasis (P