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目的观察霍奇金淋巴瘤(HRS)中IκBα蛋白、mRNA的表达特征并探讨其分子病理学机制。方法应用免疫组织化学技术观察22例HRS中IκBα蛋白的表达;应用原位杂交技术检测IκBα的mRNA表达。应用RT-PCR和直接测序法了解HRS细胞株L428、HDLM2的IκBα基因突变情况,并用Western blot测定IκBα蛋白的表达。结果(1)IκBα蛋白表达主要定位于胞质,阳性率为36.3%。所有HRS细胞IκBα蛋白表达的阳性强度较背景反应性淋巴细胞弱。(2)HRS中HRS细胞IκBα mRNA呈胞质阳性信号,阳性率为77%,杂交信号的强度高于背景反应性淋巴细胞。(3)两株HRS细胞株中,L428存在IκBα基因突变,突变位于第五外显子(C-T,DNA pos2846,cDNA pos893),形成终止密码;Western blot显示,该突变造成IκBα蛋白质羧基端截短改变。结论HRS IκBα蛋白可能存在降解水平的异常升高或结构异常。HRS来源的细胞株L428 IκBα的基因存在突变,IκBα基因突变可导致其蛋白结构异常,可能造成功能异常。
Objective To observe the expression of IκBα protein and mRNA in Hodgkin’s lymphoma (HRS) and to explore its molecular pathological mechanism. Methods Immunohistochemistry was used to observe the expression of IκBα protein in 22 cases of HRS. The mRNA expression of IκBα was detected by in situ hybridization. The mutations of IκBα gene in HRS cell line L428 and HDLM2 were detected by RT-PCR and direct sequencing. The expression of IκBα protein was detected by Western blot. Results (1) The expression of IκBα mainly located in the cytoplasm, the positive rate was 36.3%. The positive intensity of IκBα protein expression in all HRS cells was weaker than background reactive lymphocytes. (2) HRS cells HRS cells IκBα mRNA was positive for cytoplasmic signal, the positive rate was 77%, the intensity of hybridization signal was higher than background reactive lymphocytes. (3) In the two HRS cell lines, IκBα gene mutation exists in L428, and the mutation is located in the fifth exon (CT, DNA pos2846, cDNA pos893), forming a stop codon. Western blot showed that the mutation caused IκBα protein carboxy-terminal truncation change. Conclusion HRS IκBα protein may have abnormally elevated or structural abnormalities. HRS-derived cell line L428 IκBα gene mutation, IκBα gene mutation can lead to its protein structure abnormalities, may cause dysfunction.