用生物素标记的寡核苷酸探针检测入淋巴组织和淋巴瘤内免疫球蛋白轻链mRNA

DETECTION OF IMMUNOGLOBULIN LIGHT CHAIN mRNA IN HUMAN LYMPHOID TISSUES WITH BIOTINYLATED OLIGONUCLEOTIDE PROBES

本文用生物素标记的κ和λ寡核苷酸探针,在原位杂交的基础上,检测了人淋巴组织,恶性淋巴瘤和浆细胞瘤内轻链 mRNA。结果显示扁桃体和淋巴结内κ和λ轻链 mRNA 主要分布在次级淋巴滤泡的生发中心和浆细胞内,帽状区阴性。初级淋巴滤泡虽然尚无明显生发中心,但也有 mRNA阳性细胞。κ和λ轻链 mRNA 阳性细胞在淋巴组织内混合存在。相反地,淋巴瘤和浆细胞瘤内 mRNA表达则是单一型的。瘤细胞内的 mRNA 含量较多,可能与 mRNA 复制速度增快有关。实践证明经过Formalin 固定和石蜡包埋的组织切片,mRNA 仍被保存,用检测系统四步法能够捕捉到被检 mRNA,而且是特异的。

Immunoglobulin light chain mRNA in lymphoid tissues,lymphoma and plasmacy toma. was detected by using in situ hybridization with biotinylated kappa and lambda ocligonu- clotide probes.The results showed that the positive distribution of light chain mRNA in ton- sil and lymph nodes is predominantly found in germinal center of secondary follicles as well as in plasma cells,while in the mantle zone,it is completely absente The primary follicles al- so showed reactivity with Kappa and Lambda oligonucleotide probes,although they are lack of germinal centers.Both κ and λ mRNA positive cells in lymphoid tissues are concurrent and mixed.In contrast to lymphoid tissue,mRNA expression in lymphoma or plasmacytoma is restricted to monotypec mRNA.All tumor cells are stained deeply,this is probably relative to the higher copy number of light chain mRNA in tumor cells.Our results confirmed that the method is specific to localization of Kappa and Lambda light chain mRNA and showed that the specific mRNA can be detected from routine formalin-fixed and paraffinembedded section,which should be followed by four-stages method to capture the mRNA in cells.