摘要
目的观察亚高温热疗联合多西紫杉醇化疗是否具有协同抗肿瘤效应，并探讨其作用机制。方法首先采用MTT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，并筛选出有效浓度。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＋多西紫杉醇组，热疗＋多西紫杉醇组又根据热疗温度（包括39．0℃、39．5℃、40．0℃、40．5℃、41．0℃）不同细分为5个亚组。各组MCF-7细胞分别经相应干预后，利用流式细胞仪检测上述各组细胞凋亡情况及对细胞生长周期的影响。采用Westernblotting技术检测各组细胞丝裂原活化蛋白激酶（MAPK）蛋白、凋亡基因Bel-2、Bax蛋白、热休克蛋白70（HSP-70）及多药耐药基因（MDR）产物P糖蛋白（P-gP）表达情况。结果热疗联合多西紫杉醇干预可促进MCF-7细胞凋亡增加，如41．0℃热疗十多西紫杉醇组凋亡率[（37．12±6．73）％]显著高于多西紫杉醇组凋亡率[（19．16±3．42）％]；热疗联合多西紫杉醇干预可诱导MCF-7细胞生长周期停滞在G：／M期，如41．0℃热疗＋多西紫杉醇组G2／M期细胞比例[（38．18±4．72）％]显著高于多西紫杉醇组[（26．63±4．08）％]。热疗＋多西紫杉醇组MAPK通路蛋白表达较多西紫杉醇组显著增强，Bel-2蛋白表达则明显降低，而Bax蛋白表达有轻微升高。热疗十多西紫杉醇组细胞热休克蛋白70（HSP-70）、P—gP蛋白表达均较多西紫杉醇组明显增强。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（ERK）、c—Jun氨基末端激酶（JNK）、p38蛋白有关，另外亚高温热疗联合多西紫杉醇干预能增强MCF-7细胞HSP-70及P—gp蛋白表达，这可能会导致肿瘤细胞化疗耐药性产生。
Objective To confirm whether taxotere combined with mild hyperthermia will have synergistic effects, and to explore their joint mechanism of action. Methods Firstly the effective concentration of taxotere was determined by using MTT method to observe the effect of docetaxel on proliferation of human breast cancer cell line MCF-7. Then three samples of in vitro cultured human breast carcinoma MCF-7 cells were prepared, termed the the taxotere group, the taxotere plus mild hyperthermia group and the control group, and treated with effective concentration of taxotere exclusively or in combination with mild hyperthermia, or left without any special treatment. The taxotere plus mild hyperthermia group was subdivided into 5 subgroups according to the temperatures used （39.0 ℃ , 39.5℃ , 40.0℃ , 40.5 ℃ , 41.0℃ ） MTT assays were used to measure the proliferation and invasive capacity of the cells and their effective concentrations. Flow cytometry was used to detect cell apoptosis rates and any cell cycle changes in the control group. Western blotting was used to detect any changes in the expression of mitogen-activated protein kinases （MAPKs） , Bel-2/Bax, heat shock protein-70 （HSP-70） and P-gp. Results The taxotere with mild hyperthermia group demonstrated a significantly higher rate of apoptosis than that in the taxotere and control groups. There were also more cells in the GJM phase observed. Combining taxotere with mild hyperthermia was found after 24 h to have significantly increased p-ERK, p-JNK, p38, HSP-70 and P-gp protein levels and to have significantly decreased Bcl-2 protein expression in contrast with the other two groups. Conclusions Combining taxo-tere with mild hyperthermia showed synergistic effects in vitro. It seemed to be limiting the accumulation of MCF-7 cells in the G2/M phase and activating the signal pathways of MAPKs while inhibiting the activation of Bcl-2/Bax signal pathways. Combining taxotere and mild hyperthermia can accelerate the expression of HSP70 and P-gp in MCF-7
出处
《中华物理医学与康复杂志》
CAS
CSCD
北大核心
2014年第8期577-582,共6页
Chinese Journal of Physical Medicine and Rehabilitation
