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目的:探究BCL-6在经典型霍奇金淋巴瘤(classical Hodgkin’s lymphoma,cHL)细胞株LA28和过表达CD99的LA28(L428-CD99~+)中的表达差异及其意义。方法:应用免疫细胞化学和免疫荧光共聚焦检测BCL-6和CD99在cHL细胞株LA28和LA28-CD99~+中的表达;运用实时荧光定量PCR和Western blotting检测细胞株LA28和LA28-CD99~+中BCL-6和CD99的表达水平;MTT实验检测LA28和L428-CD99~+细胞增殖能力的差异;流式细胞术检测L428和LA28-CD99~+细胞的凋亡差异。结果:免疫细胞化学和免疫荧光共聚焦显示CD99在LA28细胞中为阴性表达,在L428-CD99~+细胞中为阳性表达,并定位于细胞膜;BCL-6在LA28细胞为阴性表达,在LA28-CD99~+细胞株中为阳性表达,并定位于细胞核。Western blotting显示CD99和BCL-6在LA28细胞为阴性表达,在LA28-CD99~+细胞为阳性表达。实时荧光定量PCR结果显示,与L428细胞相比,CD99和BCL-6 mRNA在LA28-CD99~+细胞中的表达量增高(P<0.01)。MTT显示LA28细胞增殖能力强于L428-CD99~+细胞(P<0.01);流式细胞术显示L428-CD99~+细胞凋亡较LA28细胞增多(P<0.01)。结论:过表达CD99诱发BCL-6表达增高,可导致L428细胞增殖能力下降及凋亡增加。
Objective: To investigate the difference and significance of BCL-6 expression in classical Hodgkin’s lymphoma (cHL) cell line LA28 and LA28 (L428-CD99 ~ +) overexpressing CD99. Methods: The expression of BCL-6 and CD99 in cLL cell line LA28 and LA28-CD99 ~ + was detected by immunocytochemistry and immunofluorescence confocal. The cell lines LA28 and LA28-CD99 ~ + were detected by real-time fluorescence quantitative PCR and Western blotting. The expression of BCL-6 and CD99 were detected by MTT assay. The proliferation of LA28 and L428-CD99 ~ + cells was detected by MTT assay. The apoptosis of L428 and LA28-CD99 ~ + cells was detected by flow cytometry. Results: The results of immunocytochemistry and immunofluorescence confocal showed that CD99 was negatively expressed in LA28 cells and positive in L428-CD99 ~ + cells, and localized in the cell membrane. BCL-6 was negative in LA28 cells, CD99 ~ + cell lines for positive expression, and located in the nucleus. Western blotting showed that CD99 and BCL-6 were negatively expressed in LA28 cells and positively expressed in LA28-CD99 + cells. Real-time PCR results showed that the expression of CD99 and BCL-6 mRNA in LA28-CD99 ~ + cells was increased compared with L428 cells (P <0.01). MTT showed that the proliferation of LA28 cells was stronger than that of L428-CD99 + cells (P <0.01). Flow cytometry showed that the apoptosis of L428-CD99 + cells was more than that of LA28 cells (P <0.01). Conclusion: The overexpression of CD99 induces the increase of BCL-6 expression, which leads to the decrease of proliferation and apoptosis of L428 cells.