应用多重标记技术对霍奇金淋巴瘤H/RS细胞属性及生物学行为的研究

目的　探索细胞凋亡相关基因、蛋白与经典型霍奇金淋巴瘤 (CHL)的相关关系及H/RS细胞的属性。方法　选用山西省肿瘤医院 6 2例存档CHL。用免疫组化ABC法检测bcl 2、CD3、CD2 0、CD30、CD15和CD10 ;免疫组化双标记技术检测RS细胞表达P5 3;ABC和DNA末端标记双标记技术检测细胞凋亡 ;免疫组化和原位杂交双标记技术检测H/RS细胞表达κRNA和λRNA ;多重标记检测背景中非肿瘤性T、B细胞的分布特点。结果　 6 2例CHL中 14例 (2 2 .5 8% )表达P5 3蛋白 ,35例 (5 6 .4 5 % )表达bcl 2蛋白。 6 2例背景非肿瘤细胞均有细胞凋亡 ,仅有 10例H/RS细胞出现凋亡 ,表达bcl 2的H/RS细胞凋亡与其bcl 2的表达呈负相关 (P =0 .0 2 )。H/RS细胞CD30全部阳性 ,4 1例表达CD15 ,8例表达CD2 0 ,6 2例均不表达CD3、MPO、bcl 6、CD10、κRNA和λRNA。非肿瘤性T细胞围绕H/RS细胞呈玫瑰花样 ,B细胞分布在其外围。结论　CHL中H/RS细胞对B系细胞标志有较大的变异。T、B淋巴细胞和H/RS细胞的分布特点可作为诊断的参考指标。多重标记可突出研究的目标细胞 ,从而有针对性的对H/RS细胞进行研究。P5 3的异常表达在CHL可能不起主要作用。bcl 2过度表达可能抑制细胞凋亡

Study on the origin of H/RS cell and their biological behavior in Hodgkin lymphoma by using multiple mark techniques

OBJECTIVE: To investigate the apoptosis-related genes and protein expression patterns in relation to classical Hodgkin lymphomas (CHL) and the origin of H/RS cell. METHODS: Sixty-two cases of CHL were retrieved from Shanxi Tumor Hospital files. An ABC method was used to detect the expression of bcl-2, CD3, CD20, CD30, CD15 and CD10, a double immunohistochemical method to study the H/RS cells P53 expression, a double immunohistochemical ABC-DNA end labeling technique to detect the apoptosis, a double immunohistochemical ABC- in situ hybridization technique to detect the expression of kappamRNA and lambdamRNA, and a multiple mark techniques to detect the distribution of background non-neoplastic T and B cells. RESULT: Of 62 CHL, 14 (22.58%) were p53 positive and 35 (56.45%) bcl-2 positive. Apoptosis was found in the background non-neoplastic cells in all of the cases, but in H/RS cells in only 10 of 62 cases. There was a significant reverse correlation between bcl-2 expression and apoptosis in H/RS cells (P = 0.02). CD30 positive H/RS cells were observed in all cases, whereas CD15 positive in only 41 cases, and CD20 positive in 8 cases. None was positive for CD3, MPO, bcl-6, CD10, kappaRNA and lambdaRNA in H/RS cells. The H/RS cells were surrounded by non-neoplastic T cells looked like a rosette and the outer periphery was B cells. CONCLUSIONS: The H/RS cell of classical Hodgkin lymphoma has a great variety of B lineage markers. The characteristic distributions of T, B and H/RS cells may serve as a reference for the diagnosis. Multiple marker technique is able to highlight the critical cells, and facilitate the study of H/RS cells. Abnormal expression of P53 may not play a major role in CHL. Over expression of bcl-2 may be linked to blockage of apoptosis in CHL.