原位杂交检测microRNA-205在乳腺癌中的表达  

In situ hybridization analysis for microRNA-205 expression in breast cancer

作　　者：范瑞琦[1] 王晟[1,2] 揭克敏[1] 熊小亮[1] 肖影群[3] 贺晓菊[4] 刘卓琦[1] 罗达亚[1] 

机构地区：[1]南昌大学基础医学院生物化学与分子生物学教研室,南昌330006 [2]杭州电子科技大学研究生院,浙江310018 [3]南昌大学附属感染医院病理科,江西330002 [4]江西省妇幼保健院,江西330005

出　　处：《中国组织化学与细胞化学杂志》2014年第1期15-19,共5页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金资助(30860319;81060054;81160248;81360313);南昌大学科研基金资助(JC200835)

摘　　要：目的探讨microRNA-205表达与乳腺恶性病变的关系。方法乳腺疾病及癌组织芯片原位杂交分析microRNA-205的表达;实时定量RT-PCR方法检测正常乳腺细胞株、恶性程度不同的乳腺癌细胞株中microRNA-205的表达。结果原位杂交分析显示,36例正常与良性乳腺病变中,33例(91.67%)表达阳性;36例乳腺癌中,23例(63.89%)表达阳性。microRNA-205的表达在乳腺正常与良性病变中的表达较恶性病变中高且有统计学差异(P=0.011),但与乳腺癌TNM分期、临床分期无关(P>0.05)。实时定量RT-PCR结果显示,四个高度恶性乳腺癌细胞株(MDA-MB-231、HS578T、BT549和SUM159PT)中microRNA-205的表达较永生化正常乳腺上皮细胞株MCF10A和四个低度恶性细胞株(MDA-MB-468、T-47D、ZR-75-1和SKBR3)中为低(P<0.05)。结论原位杂交适用于microRNA-205的表达分析;组织芯片标本原位杂交与乳腺细胞株实时定量RT-PCR分析结果提示,microRNA-205可能参与了乳腺癌的发生、发展,并随着乳腺癌的演进呈下调趋势。Objective T0 investigate the relationship between microRNA205 expression and malignant breast diseases. Methods The microRNA 205 expression assays were performed on breast diseases related tissue array with in situ hybridization （ISH）. Real time quantitative RTPCR （qRT PCR） was used to analyze the ex pression of microRA205 in normal, les-aggressive and aggressive breast cell lines. Results ISH on tissue array showed that microRNA205 was positively expressed in 33 out of 35 （91.57%） normal tissues and benign tumors and 23 out of 36 （63. 89%） malignant breast cancer, respectively. The microRNA205 expression was negatively correlated with malignancy of breast diseases （P=0. 011）, but had no association with TNM classification and clinical stages of breast cancer （P〉0.05）. The qRT PCR showed that the expressions of microRNA205 in the 4 aggressive breast cancer cell lines （MDA MB-231, HS578T, BT549 and SUM159PT） were lower than those in the normal breast epithelial cell line MCF10A and less-aggressive cell lines （MDA MB-468, T 47D, ZR75 1 and SKBR3） （P〈0.05）. Conclusion Application of ISH provides convenient conditions for microRNA205 expression detection; ISH on breast diseases related tissue array and qRT PCR analysis for breast cell lines suggest that mi croRNA205 expression is downregulated during the development and progression of breast cancer.

关 键 词：原位杂交 microRNA-205 乳腺癌 

分 类 号：R36[医药卫生—病理学]