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目的:探究经典型霍奇金淋巴瘤细胞系L428中大小细胞亚系生物学特性差异及其意义。方法:应用有限稀释法分离出L428中大小细胞亚系并进行传代培养,分别命名为L428-big和L428-small细胞。免疫细胞化学实验检测L428中大小细胞亚系CD15和CD30表达差异,MTT实验检测L428中大小细胞亚系增殖能力的差异;流式细胞术检测L428中大小细胞亚系的凋亡差异。结果:成功分离出L428细胞株中大小细胞亚系并进行传代培养;免疫组化显示L428-big细胞的CD15及CD30表达明显强于L428-small细胞;MTT实验显示,L428-big细胞和L428-small细胞之间增殖能力差异有统计学意义,F=565.273,P<0.001,L428-small细胞增殖能力高于L428-big细胞,P<0.001;流式细胞术显示,L428-big细胞的凋亡率为(3.3±0.1)%,明显多于L428-small细胞(0.4±0.1)%,P<0.001。结论:L428-small细胞增殖能力强,凋亡少,为经典型霍奇金淋巴瘤细胞起源的进一步研究提供一定的实验基础。
Objective: To investigate the differences of biological characteristics of large and small cell sublines in classical Hodgkin lymphoma cell line L428 and its significance. Methods: The large and small cell sublines of L428 were isolated and subcultured by limiting dilution method, and named L428-big and L428-small cells respectively. Differences in the expression of CD15 and CD30 in large and small cell sublines of L428 were detected by immunocytochemistry. The proliferation of large and small sublineages in L428 was detected by MTT assay. The apoptosis of large and small cell sublines in L428 was detected by flow cytometry. Results: The large and small cell sublines of L428 cell line were successfully isolated and subcultured. Immunohistochemistry showed that the expression of CD15 and CD30 in L428-big cells was significantly stronger than that of L428-small cells. MTT assay showed that L428-big cells and L428- The proliferation of L428-big cells was significantly higher than that of L428-big cells (P <0.001), F = 565.273, P <0.001. The results of flow cytometry showed that the apoptosis of L428-big cells The rate was (3.3 ± 0.1)%, significantly higher than that of L428-small cells (0.4 ± 0.1)%, P <0.001. CONCLUSION: L428-small cells have strong proliferative ability and less apoptosis, which provide some experimental basis for the further study of the origin of classical Hodgkin’s lymphoma.