三氮唑席夫碱衍生物诱导SMMC-7721细胞自相残

Triazole Schiff base derivative induces cannibalism of SMMC-7721 cells in vitro

目的分析三氮唑席夫碱衍生物(LH-37)诱导人肝癌细胞(SMMC-7721细胞)自相残的形态特征。方法1×104/ml个SMMC-7721细胞,培养在含1×10-5mol/LLH-37的培养液内24h、48h,巴氏、瑞氏染色分析细胞结构,免疫细胞化学方法观察激活型Caspase阳性细胞,透射电镜观察自相残细胞超微结构。结果常规显微镜下可见自相残的细胞表现为一个细胞被另一个细胞完全包裹,一些被邻近大细胞内在化的细胞由较大的囊泡包裹。LH-37作用SMMC-7721细胞48h,自相残细胞达5.23%,对照组为0.47%,大部分被包裹的小细胞表现为激活型Caspase-3阳性。LH-37作用24h,透射电镜下可见被大细胞包裹的小细胞呈现活细胞特征,至48h,被包裹的小细胞降解甚至只剩下残片。结论LH-37能够诱导人肝癌细胞自相残。

Objective To a nalyze the morphologic features of SMMC-7721 cannibalistic cells that induced by triazole Schiff base derivative（LH-37） in vitro. Methods The SMMC-7721 cells （1×10^4/ml）were cultured in the medium containing of 1×10^-5 mol/L LH-37 for 24h,48hThe character of cells was detected by Papanicolaou and Wright′s Staining. Immunohistochemical method was used to observe the cleaved Caspase-3 positive cells. The ultrastructure of cannibalism cells was observed by JEM 100CXII transmission electronic microscope. Results Microscopic analysis demonstrated the complete internalization of one cell within another. We noted that some cannibalistic cells in small aggregates appeared to be inside of large vacuoles, suggesting that they were internalized within a neighboring cell. The proportion of cannibalistic cells were increased after SMMC-7721 cells were cultured in the presence of LH-37 for 48 hours. The proportion of the cannibalistic cells in control and LH-37 group was 0.47% and 5.23% respectively. Many internalized cells were positive for cleaved Caspase-3 staining . Ultrastructural analysis of engulfed cells from 24 hours exhibited evidence of livecell internalization consistent with cannibalism, The most common fate for internalized cells was death after treatment with LH-37 for 48 hours, as evidenced by nuclear degradation and the eventual disappearance of some cells within the enveloping cell Conclusion The data presented indicate that LH-37 can lead to an increase of cannibalism in human hepatocarcinoma cell in vitro.