促肝细胞生长素诱导人肝癌细胞（BEL－7402）凋亡

Hepatocyte Growth-promoting Factor(pHGF) Induce Apoptosis in Proliferating Human Hepatocellualar Carcinoma Cells in Vitro

本文从细胞学、ＤＮＡ凝胶电泳、流式细胞术三方面研究促肝细胞生长素（ｐＨＧＦ）在体外对肝癌细胞增殖活性的影响。结果表明：ｐＨＧＦ对肝癌ＢＥＬ－７４０２细胞增殖有抑制作用，并存在剂量和时间相关性。其４８ｈ的半数抑制浓度（ＩＤ５０）为０．３７ｍｇ／ｍｌ±０．０４ｍｇ／ｍｌ。而３７℃灭活的ｐＨＧＦ对ＢＥＬ－７４０２细胞增殖在１５ｈ无抑制作用，在２４和４８ｈ抑制作用很弱（ＩＤ５０＞１．５ｍｇ／ｍｌ）。ＤＮＡ凝胶电泳结果表明，ｐＨＧＦ可诱导ＢＥＬ７４０２细胞产生细胞凋亡（Ａｐｏｐｔｏｓｉｓ）。流式细胞术（ＦＣＭ）结果显示：ｐＨＧＦ抑制ＢＥＬ－７４０２细胞增殖过程是先使细胞停留在Ｇ０／Ｇ１期，继而诱导细胞产生凋亡。后两项结果均显示ｐＨＧＦ对人肝癌细胞凋亡的诱导里时间和剂量相关性。

Hepatocyte growth-promoting factor(pHGF), a drug for the treatment of severe and chronic active hepatitisis in China, has been reported to accelerate hepatocyte growth in vitro and in vivo. But effect of pHGF on human hepatocelluar carcinoma cell is unknown. In the present study, we conducted an investigation to determine the anti-tumor effect of pHGF in vitro. The result showed that pHGF suppressed the proliferation of BEL-7402 cells. Inhibitory dose 50(ID50) at 48 hours of exposure to pHGF was 0.37mg/ml±0.04mg/ml. However, almost no suppressive effects were detected at 15 hours of exposure to the heat-inactived pHGF. At 24 hours and 48 hours,the ID50 of the heat-inactived pHGF were > 1.5mg/ml, less than the active pHGF(P<0.05). In agarose gel, DNA fragmentation and ladder form were demonstrated. Cell cycle analysis revealed that at the early period of exposure, pHGF induced arrest of the cells on the G0/G1 phase and at the late period, it induced apoptosis.