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目的:利用量子点(quantum dots,QDs)荧光成像技术对氯化钴(CoCl2)诱导的缺氧环境下人舌鳞癌Tca8113细胞内Heat Shock Protein 70(HSP70)蛋白进行长时间动态连续观测。方法:对人舌鳞癌Tca8113细胞分别进行常氧培养和Co Cl2模拟缺氧培养,利用QDs对细胞内HSP70蛋白进行特异性荧光标记,在激光共聚焦显微镜下观测HSP70表达及细胞内分布情况。用Image-Pro Plus软件测量缺氧0 h、6 h、12 h、24 h、36 h、48 h后HSP70蛋白的荧光信号强度变化。结果:激光共聚焦显微镜下可见人舌癌Tca8113细胞内HSP70蛋白明显表达,表现为绿色荧光。缺氧6 h后Tca8113细胞进入有丝分裂期的细胞明显增多,细胞内hsp70的表达开始增强,约在24 h时到达快速增强期,36 h组、48 h组则未见明显的进一步增强。结论:缺氧环境下Tca8113细胞有丝分裂活动明显增强,HSP70蛋白呈现出高表达,缺氧24 h时HSP70蛋白的表达达到快速增强期,随着缺氧时间的进一步延长,进入缓慢增强的平台期。
OBJECTIVE: To observe the heat shock protein 70 (HSP70) protein in human tongue squamous cell carcinoma cell line Tca8113 induced by cobalt chloride (CoCl2) under long-term dynamic observation using quantum dots (QDs) fluorescence imaging. Methods: Human tongue squamous cell carcinoma Tca8113 cells were cultured under normoxia and Co Cl2 hypoxia respectively. QDs were used to specifically fluorescently label HSP70 protein. The expression of HSP70 and the distribution of HSP70 were observed under laser scanning confocal microscope. The changes of the fluorescence intensity of HSP70 protein at 0 h, 6 h, 12 h, 24 h, 36 h, 48 h after hypoxia were measured by Image-Pro Plus software. Results: The expression of HSP70 protein in Tca8113 human tongue cancer cells was observed under confocal laser scanning microscope, showing green fluorescence. After 6 h of hypoxia, the number of Tca8113 cells entering mitosis significantly increased, the expression of hsp70 began to increase in cells, and reached a rapid increasing stage at about 24 h. No significant enhancement was observed in 36 h and 48 h groups. CONCLUSION: The mitotic activity of Tca8113 cells was significantly increased under hypoxic conditions, and the expression of HSP70 protein was highly expressed. The expression of HSP70 protein rapidly increased after 24 h of hypoxia, and reached a plateau with a slowly increasing hypoxia time.