CAJ | 학술논문

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、50.0 μmol/L)CD20单抗处理12、24、48和72 h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(50 μmol/L CD20单抗处理48 h)及对照组,使用流式细胞术测定Raji和Namalwa细胞株的凋亡率(Annexin V~+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72 h后测定细胞增殖抑制率,以50.0 μmol/L CD20单抗处理48 h后最高,为(71.91±2.05)%;采用此浓度处理48 h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.
목적 연구CD20단항대Burkitt림파류세포Bcl-2화p38MAPK단백표체적영향,탐토기억제세포증식、유도세포조망적작용궤제.방법 선용CD20(+)적Raji화Namalwa세포주,사용불동농도(12.5、25.0、50.0 μmol/L)CD20단항처리12、24、48화72 h후,용MTT법검측세포증식억제솔,이선택괄당적약물농도급작용시간;설치실험조(50 μmol/L CD20단항처리48 h)급대조조,사용류식세포술측정Raji화Namalwa세포주적조망솔(Annexin V~+화PI~-세포비례),이명학CD20단항대기조망적영향;사용Western blot법검측Raji화Namalwa세포주중 Bcl-2화p38MAPK단백표체적변화.결과 불동농도CD20단항처리Raji세포주12、24、48화72 h후측정세포증식억제솔,이50.0 μmol/L CD20단항처리48 h후최고,위(71.91±2.05)%;채용차농도처리48 h후,류식세포술검측현시,Raji세포주적세포조망솔유대조조적(2.03±0.94)%증가지(31.52±2.48)%(P＜0.05);Namalwa세포주적세포조망솔유대조조적(1.49±0.87)%증가지(21.84±1.69)%(P＜0.05).재CD20단항처리후적Raji세포주중,Bcl-2단백표체유(1.48±0.05)하강위(0.42±0.02)(P＜0.05);p38MAPK단백표체유(1.76±0.08)하강위(0.37±0.05)(P＜0.05).결론 CD20단항능직접억제림파류세포증식,통과하조Bcl-2화p38MAPK단백적표체,유도CD20(+)적Raji화Namalwa림파류세포주조망.
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(12.5,25,50 umol/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 50 uml/L Rituximab for 48h.There was statistically significant differences(P＜0.05) of each different doses and experiment 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 PI~-) 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 protein of Raji cell by Western blot shows that:compared to the control group,the expression of Bcl-2 protein was decreased markedly to(0.42±0.02) from(1.48±0.05);p38MAPK protein was decreased significantly to(0.37±0.05) from(1.76±0.08).The effect was statistically significant different(P＜0.05).Conclusion The research showed that Rituximab,an monoclonal antibody of CD20,had the effect of inhibiting the proliferation of Raji cells and inducing apoptosis in cell lines Raji and Namalwa,seem to due to down-regulation of the expression of Bcl-2 protein and p38MAPK protein.