干扰素γ与抗CD20单克隆抗体对骨髓瘤细胞增殖影响的体外研究

Study of hrγ-interferon and anti-CD20 monoclonal antibody in inhibition of myeloma cells growth in vitro

目的 :体外研究干扰素γ能否增加多发性骨髓瘤 (MM)瘤细胞表面CD2 0的表达、增强抗CD2 0单克隆抗体美罗华抗骨髓瘤的作用。方法 :将浓度为 (0～ 80 0 )U ml的重组人干扰素γ(hrγ IFN) ,联合浓度为 (0～ 2 0 )U ml的美罗华 ,分别与 14例初治 (初治组 )和 16例复发难治 (难治组 )的MM患者瘤细胞在半固体甲基纤维素培养体系中培养 ,观察它们对瘤细胞集落形成的影响 ,并用流式细胞仪测定上述不同浓度hrγ IFN处理前后瘤细胞表面CD2 0的表达。结果 :hrγ IFN浓度分别≥ 5 0U ml或≥ 10 0U ml可明显增加初治组或难治组瘤细胞表面CD2 0的表达 ;单用≥ 5 0U ml或≥ 10 0U ml的hrγ IFN对初治组或难治组瘤细胞集落形成有抑制作用 ,而单用≥ 16U ml的美罗华只对初治组瘤细胞集落形成有轻度抑制 ;≥ 5 0U ml的hrγ IFN联合≥12U ml的美罗华 ,或≥ 10 0U ml的hrγ IFN联合≥ 16U ml的美罗华 ,对初治组或难治组瘤细胞集落形成的抑制大于单用同浓度的hrγ IFN或美罗华。结论 :体外hrγ IFN能抑制MM瘤细胞集落形成 ,并可通过增加瘤细胞表面CD2 0的表达而增强美罗华对瘤细胞集落形成的抑制作用。

Objective:To observe whether γ-interferonhas could heighten level of expression of CD20 on multiple myeloma (MM) cell membrane and promote effect of anti-CD20 monoclonalantibody MabThera in suppressing myeloma cell in vitro.Methods:Colony growth of 14 untreated(UT group) and 16 relapsed or refractory(RR group) MM patients'myeloma cells were observed in the methylcellulose semisolid medium adding human recombinant γ-interferon (hrγ-IFN) of 0～800 μg/ml or MabThera of 0～20 μg/ml.Change of CD20 expression on the myeloma cells were measured after and before myeloma cells treated by hrγ-IFN.Results:CD20 expression of myeloma cells in UT group and RR group were increased after treated by hrγ-IFN when hrγ-IFN's doses were higher than 50 U/ml and 100 U/ml ; Colony growth of myeloma cells in UT group and RR group were inhibited when hrγ-IFN's doses were higher than 50 U/ml and 100 U/ml respectively, Colony growth of myeloma cells in UT group were inhibited when MabThera's doses were higher than 16 μg/ml;Suppressive effects to colony growth of myeloma cells in UT group and RR group were increased obviously when hrγ-IFN of higher than 50 U/ml or 100 U/ml combined with MabThera of 12 μg/ml or 16 μg/ml compared with alone hrγ-IFN or alone MabThera in the same doses respectively.Conclusion:Hrγ-IFN has antiproliferative effects and can evaluates CD20 expression of myeloma cell to promote MabThera antiproliferative effects to myeloma cells in vitro.