Expressions Of VEGF, EGFR And Ki-67 In Young Patients With Breast Cancer And Its Relationship With Clinicalpathology

Breast cancer is one of the most common malignant tumors of the wowen, threatening the physical and mental health seriously. In many counties,the incidence rate has the tendency to increase, which would be the second principal cause of cancer death in women worldwide.The patients of breast cancer are mosty of wowen in post-menopause in the west,but in our country are mosty in pro-menopause,which is younger than in the west. Young patients with breast cancer have diffent clinic fearures from old patients,which has easy lymph node metastasis, faster progression of disease and poor prognosis.The first probably reason is the young patients with breast cancer have a good ovaries function which making higher endogenous female hormone,it is the probably reason of early metabasis and poor prognosis.The second reason is the young patients with breast cancer are always be in progression, having higher malignancy degree.The third reason is tumor cells are always in s-dividing phase,which have higher invasion, easy metastasis and poor prognosis.The fourth reason is that young women are being in reproductive age, affiliating of pregnancy,suckling make aggravation,and the tumor is not easy to detection.The fifth reason is that young women have pyknoglandular organ,and is the predilection years of adenofibroma, hyperplasia of mammary glands, mammitis,whica make high rates of misdiagnosis and mistreatment.The sixth reason is that young women have few opportunity to receive clinical examination especially molybdenum target,which making easy missed diagnosis and late visit. VEGF is nowadays as far as known the strongest growth factor which making direct effects on endothelial cells in blood vessels, they are-playing an important role in the period of growth of tumor vessels;EGFR can accelerating the reproducing the tumor cells and revascularization, reducing apoptosis.They are relating the formation of tumor vessels und the divert of tumor; Ki-67 is the objective indicator that full disclosure the proliferative activity of the cell colony, its highly express is an important sign of proliferative active of the cells, it can reflect the proliferative activity of the cell colony. Hence the convey of VEGF, EGFR und Ki-67 in cancer tissues, the relationship from clinical stages of TNM Staging and ALNM, and furtherer discuss the characteristic of clinicopathologia of youth breast cancer are our selected analysis.OBJECTIVETo study the expressions of VEGF, EGFR and Ki-67 in young patients with breast cancer, and the correlation between their expressions and clinicopathological characteristics,so as to helping early diagnosis, specifing herapeutic regimen, thinking highly of healing;improving their survival rate.METHEDSWe retrieved 477 cases with breast cancer who were operated in Nangfang Hospital from January,2005 to December,2008 using the medical record database. We study the cases who were 35 years or under 35 years,and the cases can be operated,and all cases are pathological diagnosis as carcinoma;and who were not treated with chemotherapy,radiotherapy and endocrine secretion before operation, and the cases whose clinical data and pathologic diagnosis data was complete;and all cases were Han people;and had numbers of dissected axillary lymph nodes more than 10.Delete the cases who were recurred or had small number of dissected axillary lymph nodes (less than 10) or minority ethnic group or received with chemotherapy,radiotherapy and endocrine secretion before operation,We got the corresponding paraffin blocks from the department of pathology, then made the immunohistochemical sections by the two-step way of shortcut immuno-chemistry. Firstly, deparaffinaged the paraffin section; secondly, repaired the antigen in citrate buffer solution by microwave 25 minutes;thirdly, used the solution of peroxidase to quench the activity of endogenous peroxidase about 3 minutes in microwave, then washed the sections three times separately by Double distilled water and PBS;fourthly, the solution of antibodies to VEGF, EGFR and Ki-67 was applied and the sections were incubated at 37℃in Electric constant temperature incubator about 1 hours, then washed the sections three times by PBS; fifthly, the solution of Goat anti-rabbit HRP polymer LgG was applied and the sections were incubated at 37℃in Electric constant temperature incubator about about 30 minutes, then washed the sections three times by PBS;sixthly, used the solution of DAB to colourate about 5minutes;At last, mild Staininged with Hematoxylin after washed with water, Hydrochloride-Alcohol differentiation about 6 seconds, after dehydration by alcohol and transparent by xylene,mounted by neutral gum.We kept the solution and reaction condition identical,to ensure the unity and comparability. If the color is doubtful, we would carry out the experiment again.We got PBS instead of antibody for negative control and got the known positive reaction sections for positive control.According to the contrast coloring from negative and positive,on the assumption without false positive and false negative the tinctorial VEGF mammary cancer cell endochylemas are sepia; the tinctorial EGFR mammary cancer cell membranes and (or) endochylemas;the tinctorial Ki-67 mammary cancer cell nucleus, they have buffy pellets as positive cells, all buffy colored endochylemas are not account as positive cells, but negative cells.Select the compact district of positive cells under low power lens and count tumor cells under high power lens. The cytolymph of VEGF negative colored<25%,less control (fine particles); 25%-50% cytolymph colored, less control(fine particles with thick particles) marking(+);51%-75% cytolymph colored, middle control (thick particles) marking(++);>75% cytolymph colored, strong control (full)marking(+++).EGFR negative is control without positive or nonspecific colored;cell membrane colored is nonsequence(+) when more than 10% area has positive control;cell membrane colored is sequence without completely cell membrane is middle(++)when more than 10% area has positive control;cell membrane has sequence strong positive control with completely cell membrane is(+++)when more than 10% area has positive control.Ki-67 is negative, when positive cancer cells counts<10%,≥10% and≤25% is,,+",>25% and <50% is,,++",>50% is,,+++",>10%is positive control, und≥25% is over control.Data analyses were carried out with SPSS13.0 statistical software package. Comparison of inter-group differences were analyzed with the fourfold table or list of chi-square test line;Spearman correlation analysis was used to analyzed the relationship between the two factors.P values<0.05 were considered significantRESULTS1.According to the sixth edition of AJCC on TNM Staging and pathology typing, the young patients were 11 cases in stageⅠ,39 cases in stageⅡ,and 17 cases in stageⅢandⅣ;the control group were 21cases,252cases,137cases.The proportion of young patients in stage I were significantly higher than the old-age-group(χ2=11.741,P<0.01),The proportion of young patients in stage II had no difference const to the stageⅢandⅣ(χ2=0.256,P>0.05;χ2=1.703,P> 0.05).In the group of young patients,There were 46 cases of infitrating ductal carcinoma,8 cases of infiltrating lobular carcinoma,3cases of carcinoma medullare,7cases of mucinous adenocarcinoma.3cases of Intraductal carcinoma; In the contrast,there were 353 cases of infitrating ductal carcinoma,21 cases of infiltrating lobular carcinoma,13cases of carcinoma medullare,13cases of mucinous adenocarcinoma,4cases of Pagets carcinoma of the breast,4cases of duct carcinoma,2cases of apocrine carcinoma.There had obviously difference in pathology typing of the youth and the older,there were more cases of infitrating ductal carcinoma than the youth(P=0.000),but there were more cases of infiltrating lobular carcinoma and odther types in the youth(P=0.030,P=0.008),which had statistics significance。The ALNM rate of the youth Were 68.66%(46/67),the older were 62.20 %(255/410).The one to three ALNM rate of the youth were 20.90%,more than four were 47.76%,and the older were 34.88%,27.31%.contrast to the ALNM of one to three and more than four groups,there had significantly difference between the youth and the older(χ2=5.099,P=0.024;χ2=11.421,P=0.001),According to the more than four ALNM,there are more cases in the youth than the older,and the latter had more cases in the less than three groups.2.By using immunohistochenical method, it showed that in the young patients VEGF positive rate was 61.19%(41/67),EGFR positive rate was 64.18%(43/67), Ki-67 positive rate was 73.13% (49/67).In the older patients VEGF positive rate was 41.46%(170/410),EGFR positive rate was 40.24%(165/410), Ki-67 positive rate was 58.78%(241/410).The positive rate of VEGF, EGFR, Ki-67 were higher than the older patients significantly(P<0.05)3.There was significantly positive correlation between TNM Staging and the VEGF positive rate in the young patients and older patients (r=0.392,P=0.001;r=0.227,P=0.000),and there was significantly positive correlation between TNM Staging and the EGFR positive rate in the young patients and older patients(r=0.444,P=0.000;r=0.274,P=0.000),there was also significantly positive correlation between TNM Staging and the Ki-67positive rate in the young patients and older patients(r=0.402,P0.001;r=0.116, P=0.019);with the increase of the TNM Staging, VEGF positive rate was significantly increased in the young group and older group, and there were significant differences among the three groups (χ2=11.914,P=0.003;χ2=21.207,P=0.000);EGFR positive rate was significantly increased in the young group and older group, and there were significant differences among the three groups(χ2=13.264,P=0.001;χ2=30.871,P=0.000); Ki-67 positive rate was also significantly increased in the young group and older group, and there were significant differences among the three groups(χ2=10.837,P=0.004;χ2=18.106,P=0.000).With the increase of TNM Staging, positive rate in three groups were also rising.4:There was significantly positive correlation between ALNM and the VEGF positive rate in the young patients and older patients(r=0.360,P=0.003;r=0.288, P=0.000),and there was significantly positive correlation between ALNM and the EGFR positive rate in the youth patients and older patients(r=0.276,P=0.024; r=0.199,P=0.000),there was also significantly positive correlation between ALNM and the Ki-67 positive rate in the youth patients and older patients (r=0.343, P=0.004;r=0.422,P=0.000);With the increase numbers of the ALNM,VEGF positive rate was significantly increased in the young group and older group, and there were significant differences among the three groups(χ2=8.678,P=0.013;χ2=34.815,P=0.000);EGFR positive rate was significantly increased in the young group and older group, and there were significant differences among the three groups (χ2=6.103,P=0.047;χ2=17.052,P=0.000);Ki-67 positive rate was also significantly increased in the young group and older group, and there were significant differences among the three groups(χ2=7.985,P=0.018;χ2=83.035, P=0.000).With the numbers of the ALNM increase, positive rate in three groups were also rising.CONCLUSION1.According to distribution of the biologic characteristic young cancer patients, they have the specialties of stronger invasiveness, faster progression, earlier recrudescence diverting, and poorer prognosis.2.For its stronger invasiveness and weaker prognosis, the distinctive manifesting of youth breast cancer, which are probably concerned of the highly express of VEGF, EGFR and Ki-67 in cancer tissues.3.The measurements, which made by the convey of VEGF,EGFR und Ki-67 in cancer tissues, have certain references for the prognosis and directed therapy of breast cancer.The distribution of the biologic characteristic young cancer patients has some regular signs, which can be used as basis as multidisciplinary treatment protocols that has certain significance of predicating prognosis.The young patients, who have bad characteristic of pathobiology and biology, should have earlier diagnosis, reasonable treatment prescriptions, and systemic therapies by the clinical work, which can make better survival rate.In the researching of case of youth breast cancer, it will be more meaningful for the appraising prognosis and directing treatment, when it is not only focus on researching for ER, PR, HER-2 etc, but also the further researching of VEGF, EGFR and Ki-67.