热刺激宫颈癌Hela细胞的量效关系研究

Hyperthermia treatment of dose-response relationship study in human cervical carcinoma Hela cells

目的探讨热疗温度及加热时间对宫颈癌Hela细胞治疗的量效关系。方法将Hela细胞分为9个温度组（37～70℃），每组又分为8个时间亚组（1～30min）。分别进行水浴热疗后，倒置显微镜观察细胞形态，噻唑蓝比色法（MTT法）测定细胞增殖率，流式细胞仪测定细胞凋亡率，免疫组织化学方法测定增殖细胞核抗原（PCNA）的表达水平。结果相同温度下，随热疗时间的延长，以及在相同时间下，随着热疗温度的增加，细胞的增殖率降低，存活率降低，PCNA的表达均降低。其中，41℃、43℃组细胞无明显形态学改变，出现少量凋亡，开始对细胞有抑制和杀伤作用。46℃组，细胞出现大量圆缩，以凋亡为主要作用。50℃、55℃组坏死为主要作用。结论随着热疗温度的增高，热疗时间的延长，其对Hela细胞的治疗效果逐渐增强。热疗温度与热疗时间之问的量效关系有机结合，可达到最佳治疗效果。

Objective To investigate the dose-response relationship of the treatment temperatures and heating time on human cervical carcinoma hela cells, aiming at providing experimental evidences for clinical hyperthermia. Methods Hela cells were heated at 37 - 70℃ in temperature- controlled water baths,the temperature was divided into nine groups, each time was divided into eight subgroups （ 1 - 30 min ）. The morphology changes of cells after hyperthermia were detected by inverted microscope. Proliferation rates were measured by MTT colorimetric assay. The apoptosis rates were determined by flow cytometric analyse. The levels of proliferating cell nuclear antigen （PCNA） were measured with immunohistochemistry. Results Increaseing the heating time at the same temperature, or increaseing heating temperature at the same time, the cell proliferation, survival rates and PCNA expression decreased. There was no significant morphological change about cells ,but have small amount of apoptosis and a direct role of the suppression and destruction at 41℃ and 43 ℃ group. A large number of cells shrinked to round and a major role for apoptosis at 46℃ group. Cell necrosis was major role at 50 ℃ and 55 ℃ group. More than 55 ℃ for necrotic cells. Conclusion With the increase of heating temperature and heating time, its treatment of Hela cells gradually enhance. So combining doseeffect relationship of hyperthermia temperature and time can reach the best therapeutic effects.