应用RNA原位核酸杂交检测乳腺癌石蜡切片中ER mRNA、PR mRNA表达

DETECTION OF ER mRNA AND PR mRNA EXPRESSION IN PARAFFIN-EMBEDDED SECTIONS OF BREAST CARCINOMA BY RNA NUCLEIC ACID HYBRIDIZATION IN SITU

目的 制备地高辛标记的寡核苷酸探针 ,检测乳腺癌石蜡切片中雌激素受体 (ER) m RNA、孕激素受体(PR) m RNA表达。方法 应用 RNA原位核酸杂交 (RISH)和免疫组化 (IHC)检测技术。结果 (1) 2 78例乳腺癌石蜡切片 ER m RNA、 PR m RNA阳性率分别为 6 7.3%和 6 1.5 % ,阴性率分别为 32 .7%和 38.5 %。 ER m RNA、 PR m RNA双阳性、双阴性分别为 5 4.3%和 2 5 .6 % ,ER m RNA阳性 PR m RNA阴性或 ER m RNA阴性 PR m RNA阳性率为 2 0 .1%。 (2 )RISH法的 ER m RNA、 PR m RNA阳性率分别为 6 7.3%和 6 1.5 %均高于 IHC法 ER、 PR的 48.9%和 41.2 % (P均 <0 .0 1)。 (3) ER m RNA与 ER、 PR m RNA与 PR的阳性和阴性符合率分别为 77%和 75 .9%。 (4 )杂交信号在乳腺癌细胞中的分布可分为 ,柱状上皮的胞质的上下方、胞质内均匀或近胞膜、核周偏位和核内外分布。 (5 ) ER m RNA与 ER、 PR m RNA阳性率与组织学级别有关 , 级组中 ER m RNA的阳性率为 83.5 %分别高于 组中 6 2 .4%和 组中 5 4.5 % (P<0 .0 1) , 、 级组中 PR m RNA的阳性率为 6 6 .7%、 6 7.3% ,均高于 组中 5 0 .6 % (P<0 .0 5 )。结论 地高辛标记的 ER、 PR寡核苷酸探针和 RISH是一种较为理想的检测乳腺癌组织中 ER m RNA和 PR m RNA的分子?

? Objective Digoxingenin labelled oligonucleotide probes were prepared by direct chemical synthesis to detect the expression of estrogen receptor (ER) mRNA and progestrone receptor (PR) mRNA in paraffin embedded sections of breast carcinomas. Methods 278 cases were examined by RNA nucleic acid hybridization in situ (RISH) and immunohistochemistry assay (IHC). Results (1)The positive rates of RISH for ER mRNA and PR mRNA were 67 3% and 61 5%, and the negative rates 32 7% and 38 5%. Its phenotype: the positive rate of RISH for both ER mRNA and PR mRNA was 54 3%, the negative rate 25 6%, the ER mRNA positive and PR mRNA negative or ER mRNA negative and PR mRNA positive 20 1%. (2) The positive rates of ER and PR mRNA by RISH were higher ( P <0 01) than those of ER and PR protein by IHC which were 48 9% and 41 2% respectively.(3)The positive and negative coincidence rate of RISH and IHC for ER mRNA and PR mRNA were 77% and 75 9%. (4)Hybridization signals in breast carcinoma cells were distributed in the cytoplasma upper and lower of the columnar epithelium, but almost void in the both sides of the nuclei; identical to the distribution in breast lobuli glandulae acinar epithelial cells; mainly located in the side with more cytoplasm; in nuclei or protruded into the nuclei. (5) The positive rates of ER and PR mRNA by RISH had a close relationship to the differentiation of breast carcinomas and tissue types. The ER mRNA positive rate in group Ⅱ was 83 5%, higher ( P <0 01) than that of group Ⅰ and Ⅲ, which was 62 4% and 54 5%. The PR mRNA positive rates in group Ⅰand Ⅱ were 66 7% and 67 3%, higher than the 50 6% of group Ⅲ ( P <0 05). Conclusion Digoxingenin labelled ER, PR oligonucleotide probes and RISH could be an excellent molecular detection technique of ER mRNA and PR mRNA in paraffin embedded sections of breast carcinomas.\;〔