肝癌细胞胞内游离Ca~(2+)测定及其与间隙连接通讯功能的相关性研究
Concentrations of intracellular second masseger Ca~(2+)i in human hepatocellular carcinoma cells and the effect on gap junctional intercellular communication
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摘要: 目的 研究肝细胞肝癌和正常肝细胞胞内信号分子游离 Ｃａ２＋浓度变化，及其对间隙连接通讯功能的意义。方法 通过Ｆｌｕｏ－３ＡＭ负载和激光扫描共聚焦显微镜分析，定位及定量研究静息状态下肝癌细胞系ＨＨＣＣ，ＳＭＭＣ－７７２１和正常肝细胞系ＱＺＧ胞内游离Ｃａ２＋浓度。结合Ｌｕｃｉｆｅｒ Ｙｅｌｌｏｗ划痕标记荧光传输技术，分析对上述细胞间隙连接通讯功能的影响。结果 激光扫描共聚焦显微镜研究证实，定量分析静息状态下［Ｃａ２＋］ｉ在正常肝细胞ＱＺＧ和肝癌细胞ＨＨＣＣ，ＳＭＭＣ－７７２１中分别为１０８．３７ ｎｍｏｌ／Ｌ，３５．１３ｎｍｏｌ／Ｌ和 ４７．０８ ｎｍｏｌ／Ｌ，［Ｃａ２＋］ｉ在 ＨＨＣＣ，ＳＭＭＣ－７７２１细胞与正常肝细胞 ＱＺＧ比较显著降低。肝癌细胞ＨＨＣＣ，ＳＭＭＣ－７７２１的间隙连接通讯功能较正常肝细胞ＱＺＧ明显减弱。结论 肝癌细胞内第二信使游离Ｃａ２＋浓度的降低，与肝癌细胞的间隙连接通讯功能障碍相一致，可能与肝癌的发生密切相关。Abstract: Objective To investigate the intracellular second masseger Ca2+i in human hepatocellular carcinoma cell lines HHCC,SMMC-7721 and normal liver cell line QZG. Methods Concentrations of intra- cellular second masseger Ca2+i in cell lines were examined in laser scanning confoeal microscopy after loaded by Fluo-3 AM. Scrape loading and dye transfer showed that QZG cells hepatoma cell lines Lucifer Yellow scrape loading and dye transfer were used to detect function of gap j...
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