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第一军医大学细胞生物与医学遗传学教研室，广东 广州 ５１０５１５ 摘要目的 探讨大肠癌细胞（ＬｏＶｏ株）热休克蛋白７０（Ｈｅａｔ ｓｈｏｃｋ ｐｒｏｔｅｉｎ ７０，ＨＳＰ７０）热应激时表达规律，为进一步研究ＨＳＰ７０在ＴＮＦ－α诱导大肠癌细胞凋亡中的作用提供了实验依据。方法培养大肠癌细胞 （ＬｏＶｏ），热应激（４２℃， ３０ｍｉｎ）后分别培养２、 ５、 ８、１１ ｈ，然后运用免疫组化、Ｗｅｓｔｅｒｎ ｂｌｏｔ方法分析应激后不同时间细胞内ＨＳＰ７０的表达和分布。结果热应激２ｈ后ＨＳＰ７０合成量增加，与未应激组比有显著差异（Ｐ＜０．０５），５～８ｈ后合成量最高（Ｐ＜０．０１），１１ｈ后明显减少，与２ｈ染色情况基本相同。并观察到热应激细胞中胞核比胞质染色深，ＨＳＰ７０有核迁移现象。少量热应激死亡细胞的特异性染色更深。结论 热应激前ＨＳＰ７０在大肠癌细胞中合成较少，热应激后合成量增加，在 ５～８ ｈ达峰值后降低。
Department of Cell Biology and Medical Genetics, First Military Medical University, Guangzhou 510515, Guangdong, China Abstract Objective To investigate the expression pattern of heat shock protein 70 (Heat shock protein 70) in colorectal cancer cells (LoVo strain) -αinduced apoptosis in colorectal cancer cells provided the experimental evidence. Methods Cultured colorectal cancer cells (LoVo) were cultured in heat stress (42 ℃, 30 min) and then cultured for 2, 5, 8 and 11 h respectively. Then the expression of HSP70 in different time after stress was analyzed by immunohistochemistry and Western blot And distribution. Results The amount of HSP70 increased after 2 h of heat stress, which was significantly different from that of the untreated group (P <0.05), and reached the highest level after 5 ~ 8 h (P <0.01) Dyeing is basically the same. And observed heat-stressed cells in the cytoplasm staining deeper than the cytoplasm, HSP70 nuclear migration phenomenon. A small amount of heat-stress killed cells were more specifically stained. Conclusion HSP70 is less synthesized in colorectal cancer cells before heat stress, but increases after heat stress, reaching the peak value at 5 ~ 8 h and then decreasing.