摘要
目的 探讨热诱导 Bca CD885颊癌细胞凋亡的动力学改变及热杀伤肿瘤细胞的机理。方法 水浴加温诱导Bca CD885细胞产生凋亡。通过细胞形态观察、原位末端标记、DNA电泳及流式细胞术 DNA测定 ,从形态学、生物化学、分子生物学以及细胞学水平确定凋亡细胞特征。结果 Bca CD885细胞经 43°C40 min加热 ,呈现细胞凋亡形态改变 ,TUNEL标记见明显的凋亡小体 ,DNA电泳呈现有一定间隔的梯状条带。同时细胞周期发生了明显的特异性改变 :G0 / G1 期百分比增加 ,S期百分比下降 ,G2 / M期百分比在加热后 3 h时也发生了明显的下降。 45°C 40 min加热则出现明显的坏死改变。结论 高温杀伤 Bca CD885细胞存在细胞坏死和细胞凋亡两种方式 ,治癌温度是通过诱导细胞凋亡发挥作用。高温诱导 Bca
Objective Apoptosis is an active process of cell suicide induced by external signals and is a major mode of cell death under physilogical conditions. Treating cancers with hyperthermia may act through induction of apoptosis. So we study the apoptosis induced by hyperthermia in BcaCD885 cells, and to study the characteristics of apoptosis cells on molecular biological levels. It is possible that hyperthemia combine with gene therapeutics can be used in clinic. Method After inducing apoptosis through hyperthermia (different temperature 37°C, 43°C, 45°C and 40 min) 12 hrs, we studied the characteristics of apoptostic BcaCD885 cells on morpholgical, biochemicial, molecular biological and cellular levels with morphological observative, DNA gel electrophoresis, TUNEL staining and flow cytometry analysis. Results 1. The morphological characters of BcaCD885 cells were tested by microscopy and electron microscopy. The group of 37°C, cells growth well, the organelle in cytoplasm is intact. The goup of 43°C, cells shrink and apoptotic boby appears, endoplasm concentrain, nucleus disappear. The group of 45°C, cytonecrosis, cellorgan rupture, cytomembrance destruction. 2. TUNEL staining and DNA gel eletrophoresis. The group of 37°C, only little cell can be stained and DNA spectrum is not clear. There are a lot of staining cells and many apoptotic body in the group of 43°C. In 43°C groupp, DNA typical ladder type line spectrum was appeared. In the group of 45°C, can see mass undertint staining cells. The character is cytonecrosis. DNA spectrum is widerspreadd and patchy. 3. Flow cytometry analysis. DAN content was determined after 43°C, 40 min in 0 h, 3 h, 6 h, 12 h, 24 h, 48 h. 37°C、 40 min group was control group. By t test we found that in 0 h group DNA content was close to control group ( P >0.05), and we found there were statistical significance ( P <0.05) between others groups and control group. The population in the G 0/G 1 phase of the cell cycle significantly increased and the population in the S phase significantly decreased with a concomitant increase in apoptotic cells. Conclusion Necrosis and apoptosis are two manners to kill BcaCD885 cells. In 43°C cells dided by apoptosis mode and in 45°C by necrosis model. It is interesting that the temperature in clinic local hypoerthermic treatment cancer is 43°C. So we think that hypperthemia kill cancinoma cells by apoptotic manner. In this experment, DNA content analysis indicate that it is active death progress of carcinoma cells in hyperthermic treatment at 43°C.
出处
《口腔颌面外科杂志》
CAS
2000年第3期239-242,共4页
Journal of Oral and Maxillofacial Surgery
基金
国家自然科学基金资助 !( 396 70 788)
云南省自然科学基金资助 !( 1999. C0 0 0 7)
