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目的 :检测鬼臼乙叉甙 (VP 16 )对人结肠癌 (HIC/S)细胞增殖和凋亡的影响 ,并探讨其可能作用机理。方法 :应用MTT比色法检测VP 16对体外培养HIC/S细胞增殖的抑制作用 ,同时用DNA断片法观测VP 16对HIC/S细胞凋亡的影响。应用S P免疫组化技术对已报道的HIC/S细胞的Rb蛋白表达状态进行再证实 ,并采用Westernblot法对HIC/S细胞的Rb蛋白作半定量分析。结果 :VP 16 (0 0 45～ 2 8 12 5 μmol/L)孵育 48小时对HIC/S细胞增殖的相对抑制率为 5 0 5 2 %± 1 78%～ 91 75 %± 3 47% (P <0 0 5 ) ,IC50 为 0 0 42± 0 0 0 7μmol/L。VP 16处理HIC/S细胞 48小时能诱导细胞凋亡。S P免疫组化染色显示HIC/S细胞Rb染色为阳性 (pRb表达 ) ,VP 16有促进HIC/S细胞内Rb蛋白表达水平增高的作用。结论 :VP 16对HIC/S细胞的增殖具有抑制作用 ,并能有效地诱导HIC/S细胞凋亡。VP 16抑制HIC/S细胞增殖和诱导HIC/S细胞凋亡可能与增高细胞内Rb蛋白表达水平有关。
Objective : To detect the effect of VP 16 on the proliferation and apoptosis of human colon cancer cells (HIC/S) and to explore its possible mechanism. Methods : MTT colorimetric assay was used to detect the inhibitory effect of VP 16 on the proliferation of HIC/S cells cultured in vitro. At the same time, DNA fragmentation was used to observe the effect of VP 16 on the apoptosis of HIC/S cells. Sp immunohistochemistry technique was used to re-examine the reported Rb protein expression status of HIC/S cells, and semiquantitative analysis of Hb/S cell Rb protein was performed by Western blot. RESULTS: The relative inhibition of HIC/S cell proliferation after 48 hours of incubation with VP 16 (0 0 45 to 2 8 12 5 μmol/L) was 50 52% ± 1 78% - 91 75 % ± 3 47% (P <0 0 5 ), IC50 is 0 0 42 ± 0 0 0 7μmol/L. Treatment of HIC/S cells with VP 16 for 48 hours induced apoptosis. Sp immunohistochemical staining showed that RIC staining in HIC/S cells was positive (pRb expression), and VP 16 could promote the increase of Rb protein expression in HIC/S cells. Conclusion : VP 16 can inhibit the proliferation of HIC/S cells and can effectively induce the apoptosis of HIC/S cells. The inhibition of HIC/S cell proliferation and the induction of HIC/S cell apoptosis by VP 16 may be related to the increase of intracellular Rb protein expression.