CAJ | 학술논문

目的：探讨PI3Kδ抑制剂CAL-101对弥漫大B细胞淋巴瘤细胞系SUDHL-10和Burkitt淋巴瘤细胞系Raji的作用及其相关机制，为这类疾病的治疗提供新的思路。方法：用不同浓度的CAL-101处理Burkitt淋巴瘤细胞系Raji和弥漫大B细胞淋巴瘤细胞系SUDHL-10，以MTT法检测CAL-101对两种细胞系的增殖抑制作用；以Annexin V/PI流式细胞术和DAPI染色法检测细胞的凋亡情况；迁移实验检测淋巴瘤细胞向淋巴瘤基质细胞系HK的迁移比例；Western blot法检测CAL-101处理后淋巴瘤细胞表达磷酸化ERK的变化；MTT法结合CalcuSyn software软件分析检测CAL-101是否可协同硼替佐米抑制淋巴瘤细胞的增殖。结果：5μmol/L及更高浓度的CAL-101对Raji细胞和SUDHL-10细胞的增殖有明显的抑制作用，且呈剂量依赖性。5、10、15、20μmol/L CAL-101作用于Raji细胞48 h，细胞增殖抑制率分别为（29.17±1.23）%、（38.15±1.51）%、（46.46±1.78）%、（55.8±2.01）%，空白对照组为（1.15±0.02）%（P<0.05）。作用于SUDHL-1048 h，细胞增殖抑制率也逐渐增加（P<0.05）。CAL-101可诱导淋巴瘤细胞的凋亡，10、20μmol/L CAL-101作用于Raji细胞24 h，AnnexinV-FITC/PI双标法显示其凋亡率分别为（22.69±3.83）%和（49.96±7.36）%，均高于对照组（5.23±2.04）%（P<0.05）；作用于SUDHL-10细胞同样得到相似的结果（P<0.05）。CAL-101可显著降低淋巴瘤细胞向基质细胞的迁移，且对Raji细胞和SUDHL-10细胞的迁移率的抑制作用也呈浓度依赖性，差异均具有统计学意义（P<0.05）；Western blot检测发现CAL-101处理细胞后磷酸化ERK的表达明显降低；CAL-101与硼替佐具有协同作用，可显著抑制淋巴瘤细胞的增殖，CI<1。结论：PI3Kδ抑制剂CAL-101可抑制淋巴瘤细胞Raji和SUDHL-10的增殖，诱导凋亡，抑制其向淋巴瘤基质细胞的迁移，其机制可能通过阻断ERK信号途径而实现；CAL-101有望为侵袭性淋巴瘤的治疗带来希望。目的：探討PI3Kδ抑製劑CAL-101對瀰漫大B細胞淋巴瘤細胞繫SUDHL-10和Burkitt淋巴瘤細胞繫Raji的作用及其相關機製，為這類疾病的治療提供新的思路。方法：用不同濃度的CAL-101處理Burkitt淋巴瘤細胞繫Raji和瀰漫大B細胞淋巴瘤細胞繫SUDHL-10，以MTT法檢測CAL-101對兩種細胞繫的增殖抑製作用；以Annexin V/PI流式細胞術和DAPI染色法檢測細胞的凋亡情況；遷移實驗檢測淋巴瘤細胞嚮淋巴瘤基質細胞繫HK的遷移比例；Western blot法檢測CAL-101處理後淋巴瘤細胞錶達燐痠化ERK的變化；MTT法結閤CalcuSyn software軟件分析檢測CAL-101是否可協同硼替佐米抑製淋巴瘤細胞的增殖。結果：5μmol/L及更高濃度的CAL-101對Raji細胞和SUDHL-10細胞的增殖有明顯的抑製作用，且呈劑量依賴性。5、10、15、20μmol/L CAL-101作用于Raji細胞48 h，細胞增殖抑製率分彆為（29.17±1.23）%、（38.15±1.51）%、（46.46±1.78）%、（55.8±2.01）%，空白對照組為（1.15±0.02）%（P<0.05）。作用于SUDHL-1048 h，細胞增殖抑製率也逐漸增加（P<0.05）。CAL-101可誘導淋巴瘤細胞的凋亡，10、20μmol/L CAL-101作用于Raji細胞24 h，AnnexinV-FITC/PI雙標法顯示其凋亡率分彆為（22.69±3.83）%和（49.96±7.36）%，均高于對照組（5.23±2.04）%（P<0.05）；作用于SUDHL-10細胞同樣得到相似的結果（P<0.05）。CAL-101可顯著降低淋巴瘤細胞嚮基質細胞的遷移，且對Raji細胞和SUDHL-10細胞的遷移率的抑製作用也呈濃度依賴性，差異均具有統計學意義（P<0.05）；Western blot檢測髮現CAL-101處理細胞後燐痠化ERK的錶達明顯降低；CAL-101與硼替佐具有協同作用，可顯著抑製淋巴瘤細胞的增殖，CI<1。結論：PI3Kδ抑製劑CAL-101可抑製淋巴瘤細胞Raji和SUDHL-10的增殖，誘導凋亡，抑製其嚮淋巴瘤基質細胞的遷移，其機製可能通過阻斷ERK信號途徑而實現；CAL-101有望為侵襲性淋巴瘤的治療帶來希望。
목적：탐토PI3Kδ억제제CAL-101대미만대B세포림파류세포계SUDHL-10화Burkitt림파류세포계Raji적작용급기상관궤제，위저류질병적치료제공신적사로。방법：용불동농도적CAL-101처리Burkitt림파류세포계Raji화미만대B세포림파류세포계SUDHL-10，이MTT법검측CAL-101대량충세포계적증식억제작용；이Annexin V/PI류식세포술화DAPI염색법검측세포적조망정황；천이실험검측림파류세포향림파류기질세포계HK적천이비례；Western blot법검측CAL-101처리후림파류세포표체린산화ERK적변화；MTT법결합CalcuSyn software연건분석검측CAL-101시부가협동붕체좌미억제림파류세포적증식。결과：5μmol/L급경고농도적CAL-101대Raji세포화SUDHL-10세포적증식유명현적억제작용，차정제량의뢰성。5、10、15、20μmol/L CAL-101작용우Raji세포48 h，세포증식억제솔분별위（29.17±1.23）%、（38.15±1.51）%、（46.46±1.78）%、（55.8±2.01）%，공백대조조위（1.15±0.02）%（P<0.05）。작용우SUDHL-1048 h，세포증식억제솔야축점증가（P<0.05）。CAL-101가유도림파류세포적조망，10、20μmol/L CAL-101작용우Raji세포24 h，AnnexinV-FITC/PI쌍표법현시기조망솔분별위（22.69±3.83）%화（49.96±7.36）%，균고우대조조（5.23±2.04）%（P<0.05）；작용우SUDHL-10세포동양득도상사적결과（P<0.05）。CAL-101가현저강저림파류세포향기질세포적천이，차대Raji세포화SUDHL-10세포적천이솔적억제작용야정농도의뢰성，차이균구유통계학의의（P<0.05）；Western blot검측발현CAL-101처리세포후린산화ERK적표체명현강저；CAL-101여붕체좌구유협동작용，가현저억제림파류세포적증식，CI<1。결론：PI3Kδ억제제CAL-101가억제림파류세포Raji화SUDHL-10적증식，유도조망，억제기향림파류기질세포적천이，기궤제가능통과조단ERK신호도경이실현；CAL-101유망위침습성림파류적치료대래희망。
Objective:To detect the inhibitory effects of CAL-101, a selective inhibitor of phosphoinostitide-3'-kinase delta (PI3Kδ), on Burkitt's lymphoma cell line Raji and diffused large B-cell lymphoma cell line SUDHL-10 and elucidate its relative mechanism. Methods:Raji and SUDHL-10 cells were treated with various concentrations of CAL-101. Methyl thiazolyl tetrazolium (MTT) assay was performed to determine the inhibitory effect of CAL-101 on lymphoma cells, and cell apoptosis was measured by Annexin V/PI and DAPI staining. Migration assays were performed with transwell to detect the migration of lymphoma cells derived from the stromal cell line HK. Western blot was used to detect the phosphorylation status of the ERK pathway. MTT and CalcuSyn software analyses were preformed to detect whether or not combining CAL-101 with bortezomib induces synergistic cytoxicity. Results:CAL-101 at con-centrations of 5, 10, 15, and 20μmol/L inhibited cell proliferation in a dose-dependent manner. The proliferation rates of the Raji cells treated with 5, 10, 15, and 20μmol/L for 48 h were 29.17%± 1.23%, 38.15%± 1.51%, 46.46%± 1.78%, and 55.8%± 2.01%, respec-tively, which were significantly higher (P<0.05) than that of the control group (1.15% ± 0.02%). Similar results were found in the SUDHL-10 cells after treatment with CAL-101 (P<0.05). CAL-101 also exerted an apoptotic effect on the lymphoma cells. The apop-totic rates of the Raji cells treated with CAL-101 for 21 h were 22.69%± 3.83%and 49.96%± 7.36%, respectively, which were signifi-cantly higher (P<0.05) than that of the control group (5.23%± 2.04%). Similar results were found in the SUDHL-10 cells (P<0.05). Treatment with 5 and 10 μmol/L CAL-101 dose-dependently inhibited the migration activity of lymphoma cells to stromal cells (P<0.05). Western blot analysis showed that the expression level of ERK phosphorylation protein was significantly downregulated in the cells treated with CAL-101. A synergistic effect between CAL-101 and bortezomib was verified. That is, these two drugs can signifi-cantly inhibit the proliferation of lymphoma cells with CI values less than 1. Conclusion:The PI3Kδ-specific inhibitor CAL-101 sup-pressed the proliferation of Raji and SUDHL-10 cells, induced apoptosis, and inhibited stromal cell-derived migration. This inhibitory effect may be induced by blocking the ERK pathway. Overall, our study indicated that CAL-101 is a novel and potential agent in the therapeutic strategy against aggressive B-cell lymphoma.