CAJ | 학술논문

目的：研究不同乳腺癌细胞抗原负载的树突状细胞（DC）与细胞因子诱导的杀伤细胞（CIK）联合培养后对乳腺癌荷瘤鼠体内肿瘤的杀伤效果。方法分离乳腺癌细胞系MCF-7/ADR中的乳腺癌干细胞并制备冻融抗原，以此冲击由正常人外周血提取的单个核细胞诱导培养的DC和CIK，注入乳腺癌荷瘤鼠模型中，以此为实验组（BCSC-AP-DC+CIK组），并与普通抗原冲击的DC联合CIK组（AP-DC+CIK组）、DC+CIK组、CIK组及生理盐水组（NS组）建立对照实验。观察各组中小鼠肿瘤生长情况，采用免疫组化法检测bcl-2、bax表达，原位末端标记法（TUNEL）检测各组肿瘤组织凋亡。结果各组小鼠肿瘤体积在治疗前后及各组小鼠肿瘤体积在治疗后差异均有统计学意义（P<0.05），NS组小鼠治疗后肿瘤体积最大（3.625±0.093）cm3，BCSC-AP-DC+CIK组肿瘤体积最小（1.234±0.131）cm3。BCSC-AP-DC+CIK组bax强阳性表达，较其他组强；bcl-2弱阳性或不表达，较其他组弱。凋亡指数BCSC-AP-DC+CIK组>AP-DC+CIK组>DC+CIK组>CIK组>NS组。结论经乳腺癌干细胞抗原冲击的DC与CIK作用后诱导同种乳腺癌细胞凋亡强于经普通乳腺癌细胞抗原冲击的DC联合CIK、单纯DC与CIK共同作用后及单独CIK的治疗效果。
목적：연구불동유선암세포항원부재적수돌상세포（DC）여세포인자유도적살상세포（CIK）연합배양후대유선암하류서체내종류적살상효과。방법분리유선암세포계MCF-7/ADR중적유선암간세포병제비동융항원，이차충격유정상인외주혈제취적단개핵세포유도배양적DC화CIK，주입유선암하류서모형중，이차위실험조（BCSC-AP-DC+CIK조），병여보통항원충격적DC연합CIK조（AP-DC+CIK조）、DC+CIK조、CIK조급생리염수조（NS조）건립대조실험。관찰각조중소서종류생장정황，채용면역조화법검측bcl-2、bax표체，원위말단표기법（TUNEL）검측각조종류조직조망。결과각조소서종류체적재치료전후급각조소서종류체적재치료후차이균유통계학의의（P<0.05），NS조소서치료후종류체적최대（3.625±0.093）cm3，BCSC-AP-DC+CIK조종류체적최소（1.234±0.131）cm3。BCSC-AP-DC+CIK조bax강양성표체，교기타조강；bcl-2약양성혹불표체，교기타조약。조망지수BCSC-AP-DC+CIK조>AP-DC+CIK조>DC+CIK조>CIK조>NS조。결론경유선암간세포항원충격적DC여CIK작용후유도동충유선암세포조망강우경보통유선암세포항원충격적DC연합CIK、단순DC여CIK공동작용후급단독CIK적치료효과。
Objective To investigate the tumor-inhibitory effect of cytokine-induced killer cells(CIK)co-cul-tured with dendritic cells (DC)pulsed by breast cancer stem cell antigen on the same tumor-bearing mice. Methods Breast cancer stem cells were isolated from the cell line of MCF-7/ADR and extract lyses antigen of the stem cell was saved. DC and CIK derived from peripheral blood mononuclear cells of healthy individuals were co-cultured and pulsed or un-pulsed by the above antigen lyses. This DC+CIK were injected to breast tumorbearing mice (BCSC-AP-DC+CIK group), and were used to compared with the common breast cancer cell antigen (rather than breast cancer stem cell antigen) pulsed DC+CIK group(AP-DC+CIK group), DC+CIK group, CIK CIK group and normal saline group(NS group). The tumor-inhibitory effect were evaluated and compared among all 5 groups through the tumor size, TdT-mediated dUTP nick end labeling test (TUNEL), examining expression level of bcl-2 and bax by immunohistochemistry. Results The tumor size in each group before and after therapy and the tumor size after therapy between each group was of significant difference(P<0.05). The maximum size is NS group(3.625±0.093)cm3 and BCSC-AP-DC+CIK group is minimum,which is (1.234±0.131)cm3. BC-SC-AP-DC+CIK group is of highest expression of bax and apoptotic index value, lowest bcl-2 expression in all 5 groups. Conclusion The CIK co-cultured with DC pulsed breast cancer stem cell antigen was more effective to induce apoptosis of breast cancer cells than those of CIK cells co-cultured with DC pulsed breast cancer cell antigen,CIK cells co-cultured with DC and CIK cells.