CAJ | 학술논문

目的：探讨亚高温热疗联合多西紫杉醇对人乳腺癌增殖的影响及其相关机制。方法采取四甲基偶氮唑盐法观察多西紫杉醇对人乳腺癌细胞株 MCF‐7细胞增殖的影响，且筛选出有效浓度。将体外培养的 MCF‐7细胞分为空白对照组、多西紫杉醇组及热疗与多西紫杉醇联合组，热疗与多西紫杉醇联合组依据不同温度（39．0℃、39．5℃、40．0℃、40．5℃、41．0℃）分为5个亚组。对各组细胞予以相应干预后，采取流式细胞仪检测各组细胞凋亡状况及细胞生长周期变化。结果随着热疗温度上升，细胞凋亡比例渐渐增高，41．0℃热疗时凋亡率[（38．90±6．88）％]最高，显著高于单一应用多西紫杉醇与其他各亚组；c‐Jun 氨基末端激酶在热疗温度为41．0℃时达到最高值，而细胞外调节蛋白激酶1／2、p‐p38蛋白表达在热疗温度为40．0℃达到最高值，同其余亚组相比，差异均有统计学意义（P＜0．05）；热疗与多西紫杉醇联合等亚组 G2／M 期细胞比例明显多于单一使用多西紫杉醇组，抑制 MCF‐7细胞周期进程的效应明显强于多西紫杉醇，且随着热疗温度增加，G2／M 期细胞比例增高，差异有统计学意义（P＜0．05）。热疗与多西紫杉醇联合组丝裂原活化蛋白激酶通路蛋白表达较多西紫杉醇组显著增强，Bcl‐2蛋白表达则明显降低，而 Bax 蛋白表达有轻微升高。结论亚高温热疗联合多西紫杉醇干预能抑制人乳腺癌细胞增殖，诱导细胞凋亡，具有协同抗肿瘤效应，其治疗机制可能与激活细胞外调节蛋白激酶、c‐Jun 氨基末端激酶、p38蛋白有关。
목적：탐토아고온열료연합다서자삼순대인유선암증식적영향급기상관궤제。방법채취사갑기우담서염법관찰다서자삼순대인유선암세포주 MCF‐7세포증식적영향，차사선출유효농도。장체외배양적 MCF‐7세포분위공백대조조、다서자삼순조급열료여다서자삼순연합조，열료여다서자삼순연합조의거불동온도（39．0℃、39．5℃、40．0℃、40．5℃、41．0℃）분위5개아조。대각조세포여이상응간예후，채취류식세포의검측각조세포조망상황급세포생장주기변화。결과수착열료온도상승，세포조망비례점점증고，41．0℃열료시조망솔[（38．90±6．88）％]최고，현저고우단일응용다서자삼순여기타각아조；c‐Jun 안기말단격매재열료온도위41．0℃시체도최고치，이세포외조절단백격매1／2、p‐p38단백표체재열료온도위40．0℃체도최고치，동기여아조상비，차이균유통계학의의（P＜0．05）；열료여다서자삼순연합등아조 G2／M 기세포비례명현다우단일사용다서자삼순조，억제 MCF‐7세포주기진정적효응명현강우다서자삼순，차수착열료온도증가，G2／M 기세포비례증고，차이유통계학의의（P＜0．05）。열료여다서자삼순연합조사렬원활화단백격매통로단백표체교다서자삼순조현저증강，Bcl‐2단백표체칙명현강저，이 Bax 단백표체유경미승고。결론아고온열료연합다서자삼순간예능억제인유선암세포증식，유도세포조망，구유협동항종류효응，기치료궤제가능여격활세포외조절단백격매、c‐Jun 안기말단격매、p38단백유관。
Objective To explore the synergistic effect of docetaxel plus mild hyperthermia on the proliferation of human breast carcinoma cells and its mechanism .Methods M T T assay was used to test the inhibitory effect of do‐cetaxel on the proliferation of human breast carcinoma MCF‐7 cells ,and the effective concentration of docetaxel was screened .The human breast carcinoma MCF‐7 cells cultured in vitro were divided into blank control group ,docetaxel group and docetaxel plus hyperthermia group .The docetaxel plus hyperthermia group was divided into 5 subgroups (39 .0 ℃ group ,39 .5 ℃ group ,40 .0 ℃ group ,40 .5 ℃ group and 41 .0 ℃ group) ,according to the different temper‐atures of hyperthermia .Flow cytometry was used to detect the cell apoptosis and the change of cell growth cycle in every group after intervention .Results The apoptosis rate of cells increased with the temperature of hyperthermia . 41 .0 ℃ group had the highest apoptosis rate of cells [(38 .90 ± 6 .88)% ] ,which was significantly higher than do‐cetaxel group and other subgroups of docetaxel plus hyperthermia group .The activity of c‐Jun amino‐terminal kinase reached maximum when the hyperthermia temperature was 41 .0 ℃ ,while the activities of ERK1/2 and p‐p38 reached maximum when the hyperthermia temperature was 40 .0 ℃ ,which was significantly different from other subgroups of docetaxel plus hyperthermia group(P< 0 .05) .The proportions of cells at G2 /M phase in the subgroups of docetaxel plus hyperthermia group were significantly higher than docetaxel group .The inhibitory effect of docetaxel plus hyper‐thermia on the MCF‐7 cell cycle was stronger than docetaxel alone .The proportion of cells at G2 /M phase increased with the hyperthermia temperature ,with statistical difference(P< 0 .05) .Compared with docetaxel group ,the expres‐sion level of mitogen activated protein kinase pathway protein in docetaxel plus hyperthermia group was significantly higher ,the expression level of Bcl‐2 significantly reduced ,and the expression level of Bax slightly increased .Conclu‐sion Docetaxel plus hyperthermia could inhibit the proliferation of human breast carcinoma cells and induce the ap‐optosis ,which showed a synergistic antitumor effect .The mechanism might relate with the activation of extracellular regulated protein kinase ,c‐Jun amino‐terminal kinase and p38 .