Expression Of HDAC1,HDAC2 And HDAC3 Protein In Intraductal Proliferative Lesions Of The Breast, Tumor Molecular Subtypes And Associations With Clinicopathological Feature

Background and Goals: The incidence of breast cancer was highly malignant threat to women,s health, in the recent years with the steroid hormone antagonists(tamoxifen etc.) and for the HER2 gene targeted therapy(trastuzumab) was used, so that part of the breast cancer patients from treatment, effectively prolong the survival of the patients, but for hormone receptor(ER,PR) test negative, no amplification of HER2 gene case, namely “three patient with negative” breast cancer, current treatment was still unsatisfactory. Histone acetylation and deacetylation as activation and inhibition makers of gene transcription, in recent years, was considered to play an important role in the development of the occurrence of tumor, with the histone deacetylase inhibitor(HDACIs) come out, It has been proposed so-called “transcription therapy” concept, providing a new therapeutic approach foe the treatment of malignant tumors, This study will explore HDACS in gene transcriptional level the possible mechanism to promote the development of breast cancer, And provide a theoretical basis for a new treatment method and choose the best treatment for breast cancer patients.Materials and Methodology:1、During the period from January 2010 to June 2014 in North Sichuan Medical College Hospital Department of pathology the breast malignant tumor paraffin pathology samples seized the archive in 531 cases with complete clinical data screening in invasive ductal carcinoma(IDC) 278 cases, Randomly selected 33 cases of breast ductal carcinoma in situ(DCIS), 5 cases of breast atypical ductal hyperplasia(ADH), 6 cases of breast usual ductal hyperplasia(UDH), And fibroadenoma(FA) of non-proliferative ductual as normal controls.2、HDAC1, HDAC2 and HDAC3 protein expression was detected using immunohistochemistry in intraductal proliferative lesions of the breast(UDH-ADH-DCIS-IDC) in different stages of the sample Molecular typing of tumor, Discuss the change rule and the clinical pathological significance. HDAC1, HDAC2 and HDAC3 protein expression was detected using Western blot method in fresh breast cancer, Combined with IHC IDC paraffin sample test results, To discusses the protein and clinical pathological indicators as well as the significance of tumor molecular subtype. Mean ± standard deviation relative to the average gray value of protein, protein expression between different groups using paired T-test and chi-square test; Kaplan-Meter survival analysis using Log-rank test method, using Cox proportional hazards regression model, univariate and multivariate analysis of prognostic factors. Disease-free survival in lesions imaging examination found that patients with recurrence or distant metastasis lesions to end time, to death for patients with overall survival event destination.Results:1、Clinical date: among 278 cases of breast cancer, 50 years old is the median age, 49 years is the average age(31-82years), 276 cases with breast mass(99.2%) as initial symptoms, 2 cases were nipple discharge or hemorrhage; 152 cases(54.7%) were the left breast, 126 cases(45.3%) were the right breast; 111 cases(39.9%) are the upper outer quadrant of the breast, it was the most common primary site, the average diameter of the tumor was 3.6cm(1.0-10.0); 174 cases(62.6%) with axillary lymph node metastasis, metastatic lymph nodes of 1-31 gold Unequal, 7 cases of distant metastases(2.5%), involving 2 cases with bone metastasis, 2 cases with lung metastasis, 3 cases with liver metastasis. Histological grade: 17 case are Grade I(6.1%), 194 case are grade II(69.8%), 67 cases are grade III(24.1%). 125 cases are followed up(45%),, follow-up 9 deaths(7.2%), 116 cases of living(92.8%)(5 cases for survival with tumor).2、Immunohistochemistry results: In FA, UDH, ADH, DCIS, the positive rate of HDAC1 protein were 45%, 100%, 100%, 90.9%; the positive rate of HDAC2 protein were 50%, 100%, 100%, 81.8%; the positive rate of HDAC3 protein were 90.0%, 83.3%, 80.0%, 66.7%. Among 278 cases of breast cancer, 247 cases(88.8%) are expression of HDAC1, 235 cases(84.5%) are expression of HDAC2, 201 cases(72.3%) are expression of HDAC3, three the average positive expression rate of tumor tissues are 66.7%,54.3%, 52.8%. 176 cases(63.3%) are expression of ER, 158 cases(56.8%) are expression of PR, 179 cases(64.4%) are expression of HER2, 204 cases(73.4%) are highly expression of ki67(≥14%).3、Western-blot results: The positive expression of HDAC1 in HR(ER /PR) positive group was higher than negative group(P<0.05). The positive expression of HDAC1 in Ki67 high expression group was higher than Ki67 low expression group(P<0.05). The positive expression of HDAC2 in HER2 positive group was higher than negative group, there was significant difference(P<0.05). The positive expression of HDAC3 in HER2(+) subtype was significantly higher than basal-like subtype(P<0.05).4、Molecular classification: 7 cases(2.5%) were of Luminal A subtype, 63 cases(22.7%) Luminal B subtype(HER2-), 113 cases(40.6%) Luminal B subtype(HER2+), 66 cases(23.7%) HER2(+) subtype, and 29 cases(10.4%) basal-like subtype.5、Statistical results5.1 The positive expression of HDAC1 and HDAC2 protein in UDH、ADH、DCIS、IDC was significantly higher than that of FA(P<0.05), While the positive expression of HDAC3 protein in DCIS、IDC was significantly lower that of FA(P<0.05), Between FA and UDH、ADH, The positive expression of HDAC3 protein was not significant difference. HDAC1/2/3 protein in the lesions of UDH-ADH-DCIS- IDC development process only between ADH and DCIS, the expression have significant difference(P<0.05), between UDH and ADH as well as DCIS and IDC, the expression of the three rate showed no significant difference(P>0.05).5.2 HDAC1/2/3 protein in Luminal A subtype was higher than other subtypes(P<0.05). The positive rate of HDAC3 protein in HER2(+) subtype was significantly higher than basal-like subtype(P<0.05). But the expression of HDAC1/2 protein was not significant difference(P>0.05). Between Luminal B subtype(HER2-) and Luminal B subtype(HER2+), the expression of HDAC1/2/3 protein was not significant difference(P>0.05). Between Luminal B subtype and HER2(+) subtype or basal-like subtype,the expression of HDAC1/2/3 protein was not significant difference(P>0.05)5.3 The expression of HDAC1 and the expression of ER、PR、Ki67 have a positive correlation. The expression of HDAC2 and the expression of HER2 have a positive correlation. The expression of HDAC1 in low-grade tumors was significantly higher than high-grade tumors, The expression of HDAC1 in high-grade tumors was significantly higher than low-grade tumors, But HDAC2 protein was not correlated with history grade of breast cancer.The expression of HDAC1、HDAC2 and HDAC3 protein was not significantly correlated with the breast cancer patient age, the tumor size, lymph node status..5.4 Survival analysis: the low expression of HDAC1 protein, the number of lymph node metastasis was negatively correlated with postoperative the disease-free survival and overall survival(P<0.05); Multi-factors Cox regression analysis showed: the number of lymph node metastasis was independent risk factors for prognosis of IDC(P<0.05).Conclusions:1、The expression of HDAC1/2/3 protein in breast hyperplastic lesions spectrum is higher than that of non proliferative ductal epithelium, in the expression of HDAC3 is lower than that in non proliferative ductal epithelium, indicating that HDAC1/2 has a certain role in the occurrence of breast cancer, HDAC3 has certain protective effect on breast.2 、 The full expression of HDAC1/2 in UDH, ADH disease, the expression was down-regulated in DCIS, IDC, the turning point may be the key to breast intraductal proliferative lesions of carcinogenesis, Suggest that changes in histone acetylation levels is an important early event in the development and progression of breast cancer.3、The expression of HDAC1 protein and ER、PR、Ki67 have a certain synergy in IDC, the high expression of HDAC1 indicates a good prognosis, postoperative the disease-free survival and overall survival extended, therefore, we speculate on the possible epigenetic mechanisms of breast cancer is: HDAC1 can up regulate the expression level of estrogen receptor and Ki67 protein, improving the sensitivity of postoperative hormone receptor antagonists and chemotherapy, so as to prolong the survival of postoperative.4、The expression of HDAC1/2/3 is correlated with the molecular subtypes of breast cancer, especially the highest expression being observed in luminal A subtype; ER and PR are negative expression in breast cancer, HDAC3 has play a more significant role in promoting the occurrence and development of the amplification of HER2 gene of breast cancer.