CD20单抗对Burkitt淋巴瘤细胞Bcl-2和p38MAPK蛋白表达的影响  

A Study on Mechanism of Expression of Bcl-2 and p38MAPK Protein Decreased by Rituximab in Burkitt Lymphoma Cell Line

作　　者：秦文娇[1] 顾静文[1] 许小平[1] 陈洁[1] 陈波斌[1] 林果为[1] 

机构地区：[1]复旦大学附属华山医院,上海市200040

出　　处：《中国全科医学》2010年第3期251-253,共3页Chinese General Practice

基　　金：卫生部卫生技术评估重点实验室开放基金

摘　　要：目的研究CD20单抗对Burkitt淋巴瘤细胞Bcl-2和p38MAPK蛋白表达的影响,探讨其抑制细胞增殖、诱导细胞凋亡的作用机制。方法选用CD20(+)的Raji和Namalwa细胞株,使用不同浓度(12.5、25.0、50.0μmol/L)CD20单抗处理12、24、48和72h后,用MTT法检测细胞增殖抑制率,以选择适当的药物浓度及作用时间;设置实验组(50μmol/LCD20单抗处理48h)及对照组,使用流式细胞术测定Raji和Namalwa细胞株的凋亡率(AnnexinV+和PI-细胞比例),以明确CD20单抗对其凋亡的影响;使用Western blot法检测Raji和Namalwa细胞株中Bcl-2和p38MAPK蛋白表达的变化。结果不同浓度CD20单抗处理Raji细胞株12、24、48和72h后测定细胞增殖抑制率,以50.0μmol/LCD20单抗处理48h后最高,为(71.91±2.05)%;采用此浓度处理48h后,流式细胞术检测显示,Raji细胞株的细胞凋亡率由对照组的(2.03±0.94)%增加至(31.52±2.48)%(P<0.05);Namalwa细胞株的细胞凋亡率由对照组的(1.49±0.87)%增加至(21.84±1.69)%(P<0.05)。在CD20单抗处理后的Raji细胞株中,Bcl-2蛋白表达由(1.48±0.05)下降为(0.42±0.02)(P<0.05);p38MAPK蛋白表达由(1.76±0.08)下降为(0.37±0.05)(P<0.05)。结论CD20单抗能直接抑制淋巴瘤细胞增殖,通过下调Bcl-2和p38MAPK蛋白的表达,诱导CD20(+)的Raji和Namalwa淋巴瘤细胞株凋亡。Objective To study the inhibitory effect of Rituximab on Burkitt lymphoma cell line and the apoptosis - inducing activity by decreasing Bcl -2 and p38MAPK protein. Methods selecting human Burkitt lymphoma cell lines Raji and Namalwa, which were CD20 positive, using Dimethyl Sulfoxide to detect the growth rate among different Rituximab concentration groups （12. 5, 25, 50umol/L） and different time groups （12, 24, 48, 72h）, in order to choose proper drug concentration and action time. According to above result, establish control and experimental group （50 umol/L for 48h）. Apoptosis rates of cell lines Raji and Namalwa were measured with flow cytometry （FCM）. Expression of the level of Bcl -2 and p38MAPK protein of both Raji and Namalwa lymphoma cells were detected by Western blot. Results The growth rate results ： After treated with Rituximab for 12h, 24h, 48h to 72h, the best inhibitory rate of Raji lymphoma cells was （71.91 -± 2. 05）% when treatment with 50uml/L Rituximab for 48h. There was statistically significant differences （ P 〈 0.05 ） of each different doses and experi- ment time shows that Rituximab could inhibit the proliferation of Raji cells in a dose - and time - dependent manner. After treat with 50umol/L Rituximab for 48h, the number of earlier apoptotic Burkitt lymphoma cells （ Annexin V ± and Pf - ） was increased gradually compared to the control group. The apoptosis rate of Raji lymphoma cells was （31.52 ± 2. 48 ） %, increased gradually compared to the control group （2. 03 ± 0. 94） %. And the Namalwa lymphoma cells was （21.84 ± 1.69 ） % , also increased gradually compared to the control group （ 1.49 ± 0. 87 ） %. That was Rituximab could induce significant increasing in apoptosis rate of lymphoma cells. In addition, after treated with Rituximab, analysis on expression of Bcl -2 protein and p38MAPK pro- tein of Raji cell by Western blot shows that ： compared to the control group, the expression of Bcl - 2 protein was decreased markedly to （0.4

关 键 词：CD20单抗 凋亡 BCL-2蛋白 p38MAPK蛋白 

分 类 号：R733.4[医药卫生—肿瘤]