Establishment Of Human Breast Cancer Tissue Resource Bank

Object:Breast cancer is one of the most frequently seen malignant tumors in female. In China, there is all increasing tendency of breast cancer among younger women, and the incidence of breast cancer is rising up. Along with research of molecular biology of breast cancer, the demand for tissue samples grows day by day, especially after the coming of individual medicine era. The formerly researchers collect tumor specimen only according to the need of their specific research topic, many valuable tumor specimens and related data are lost, resulting in the large lost of further study of breast cancer. This study aims to establish a human breast cancer tissue bank so that to provide high quality tissue specimens and patient data for the research of preclinical medicine,clinical medicine and molecular biology on breast cancer. Methods:(1) To develop an operation process chart for the construction of tumor bank and a serious of standard operating procedures (SOP) for each technology factor for quality control. (2) Breast cancer samples from operative patients were collected immediately after surgical removal and subdivided into aliquots. One was fixed, embedded, sliced and made into paraffin block step by step by pathologist, followed by pathology check such as hematoxylin and eosin (HE) examination and immunohistochemistrical assessment. The pathologist evaluated positive and negative rate of estrogen receptor (ER), progesterone receptor (PR), CerbB-2/HER 2, which are specific biochemical indicators of breast cancer. A paraffin-embedded breast cancer tissue bank was established by storing and evaluating these paraffin stocks. (3) The remaining tissue, approximately 0.5cm3 thickness, was placed into a plastic disposable vial labeled with the specimen number, person ID number and stored at-80℃freezer. In some patients 4-6 vials (aliquots) were collected per patient. One small piece of tissue was used for DNA, RNA and protein extraction and the quality were evaluated by spectrophotometric method and electrophoresis. Fresh breast cancer tissue bank and breast cancer biology molecular database were established by storing tumor tissue and DNA/RNA/Protein at-80℃freezer. (4) 5ml patient's blood was collected respectively and well frozen at-80℃. (5) By designing a series informational tables, we could collect a lot of information on patients, including clinical, pathological and basic research information (biomarker), and established breast cancer information database. On the other hand, we could collect the previous history and related informations of breast cancer patient by filling "Women Breast Health Survey Form", which would provide a reliable support for future large-scale epidemiological screening for breast cancer. Results:(1) A operation process chart for breast cancer tissue bank construction and a series of SOP, including standarded bar code label, DNA, RNA, protein extraction SOP, and blood sample collection SOP were made. (2) HE staining showed that the operational tissue was breast cancer, and immunohistochemical staining showed that ER, PR positive particles were in the cell nucleus, CerbB-2/HER2 positive particles were on cell membrane, all these biology moleculars were yellow, brown or yellow-brown particles. (3) The OD260/OD280 of DNA extracted from breast cancer tissue was 1.8-2.0, and the map of agarose gel electrophoresis of RNA showed two bands of 18s and 28s, the intensity of 28s RNA was 1.5-2 times of 18s RNA. All these data showed that high quality control of the breast cancer tissue. (4) The OD260/OD280 of DNA in blood cells of breast cancer patient were between 1.8-2.0, and the electrophoresis result of serum protein showed a series of clear protein bands, indicating that the quality of frozen blood samples was good. (5) Several information tables and "Women Breast Health Survey " were obtained. Conclusion:Framework of human breast cancer tissue bank construction was preliminary established. Breast cancer samples from 110 patients were collected. The cellular and molecular biology assessment indicates high quality control of the samples indicating that samples in this tumor bank can meet the requirments of use in basic, clinical, and/or biomarker cancer research studies.