CAJ | 학술논문

目的：研究汉防己甲素( tetrandrine, Tet)抑制结肠癌细胞增殖作用及其可能的分子机制。方法采用结晶紫染色和流式细胞术检测不同浓度Tet对结肠癌LoVo细胞增殖的抑制作用。采用流式细胞术分析Tet对LoVo细胞凋亡的促进作用。利用Western blot检测Tet对IGFBP-5的表达影响。采用荧光素酶报告质粒,IGFBP5以及IGFBP5 siRNA的重组腺病毒,研究Tet抑制结肠癌细胞增殖的分子机制。结果 Tet能明显抑制LoVo细胞增殖,并能诱导G1期阻滞,同时促进LoVo细胞凋亡。 Tet呈浓度依赖性降低LoVo细胞中IGFBP5的表达水平。与对照组相比, Tet明显抑制TOP-luc报告质粒的转录活性,外源性过表达IGFBP5能部分逆转这种抑制作用；Tet明显下调LoVo细胞中c-Myc表达水平,外源性过表达IGFBP5能部分逆转,但沉默IGFBP5表达不能逆转Tet对c-Myc表达水平的抑制作用。结论 Tet能抑制结肠癌细胞LoVo的增殖,并诱导凋亡,这种作用可能与Tet通过下调 IGFBP5表达进而抑制 Wnt/β-catenin 信号有关。
목적：연구한방기갑소( tetrandrine, Tet)억제결장암세포증식작용급기가능적분자궤제。방법채용결정자염색화류식세포술검측불동농도Tet대결장암LoVo세포증식적억제작용。채용류식세포술분석Tet대LoVo세포조망적촉진작용。이용Western blot검측Tet대IGFBP-5적표체영향。채용형광소매보고질립,IGFBP5이급IGFBP5 siRNA적중조선병독,연구Tet억제결장암세포증식적분자궤제。결과 Tet능명현억제LoVo세포증식,병능유도G1기조체,동시촉진LoVo세포조망。 Tet정농도의뢰성강저LoVo세포중IGFBP5적표체수평。여대조조상비, Tet명현억제TOP-luc보고질립적전록활성,외원성과표체IGFBP5능부분역전저충억제작용；Tet명현하조LoVo세포중c-Myc표체수평,외원성과표체IGFBP5능부분역전,단침묵IGFBP5표체불능역전Tet대c-Myc표체수평적억제작용。결론 Tet능억제결장암세포LoVo적증식,병유도조망,저충작용가능여Tet통과하조 IGFBP5표체진이억제 Wnt/β-catenin 신호유관。
Aim To investigate the anti-proliferating effect of tetrandrine ( Tet ) on colon cancer cells and its possible molecular mechanism. Methods We intro-duced crystal violet staining and flow cytometry to ana-lyze the effect of Tet on proliferation in LoVo cells. Flow cytometry was used to detect the effect of Tet on apoptosis in LoVo cells. Western blot assay was taken to analyze the effect of Tet on the expression of insulin-like growth factor binding protein 5 ( IGFBP5 ) . Final-ly, luciferase reporter assay, recombinant adenovirus mediated over-expression or silence of IGFBP5 were used to analyze the possible role of IGFBP5 in the anti-proliferating effect of Tet on colon cancer cells. Re-sults Crystal violet staining and flow cytometery anal-ysis results showed that Tet could exert an anti-prolifer-ating effect and induce apoptosis in LoVo cells. Tet de-creased the expression of IGFBP5 in a concentration-dependent manner. Tet inhibited the transcriptional ac-tivity of pTOP-luc reporter, which could be reversed by exogenous expression of IGFBP5 mostly. Similar results were found in the expression of c-Myc, but IGFPB5 knockdown couldn’ t reverse this effect. Conclusion Tet can inhibit the proliferation of colon cancer cells, and this effect may be mediated by down-regulating the expression of IGFBP5 to inhibit Wnt/β-catenin signa-ling transduction partly.