CAJ | 학술논문

目的：通过对经初步筛选证明具有生物活性的查尔酮类似物A59可能涉及的信号通道进行检测，进一步明确其抗肿瘤活性及作用机制。方法：对于以查尔酮为先导物设计合成类似物，以MTT法检测其对于人结肠癌细胞（HCT-116）的杀伤作用，筛选出具有较好生物活性的化合物A59。通过FCM法检测A59诱导HCT-116细胞凋亡和抑制细胞周期的情况。Western blot法检测HCT-116细胞中A59对ATF4及CHOP表达的影响。利用siRNA技术敲除CHOP后，用免疫荧光和FCM法检测在A59对于细胞凋亡的影响。结果：通过对自行设计合成查尔酮类似物进行筛选，发现A59对于HCT-116具有较强的增殖抑制作用（IC50=7.6μmol/L）。通过FCM法检测发现A59是通过诱导HCT-116细胞发生凋亡并抑制细胞周期发挥其生物活性。通过Western blot法检测发现A59剂量依赖性激活内质网应激信号通路中的关键蛋白CHOP，并通过siRNA基因沉默技术进一步确证A59是通过刺激CHOP的过表达发挥促进细胞凋亡的作用。结论：查尔酮类似物A59通过激活内质网应激信号通道中的关键蛋白CHOP来发挥对CHT116的增殖抑制作用。
목적：통과대경초보사선증명구유생물활성적사이동유사물A59가능섭급적신호통도진행검측，진일보명학기항종류활성급작용궤제。방법：대우이사이동위선도물설계합성유사물，이MTT법검측기대우인결장암세포（HCT-116）적살상작용，사선출구유교호생물활성적화합물A59。통과FCM법검측A59유도HCT-116세포조망화억제세포주기적정황。Western blot법검측HCT-116세포중A59대ATF4급CHOP표체적영향。이용siRNA기술고제CHOP후，용면역형광화FCM법검측재A59대우세포조망적영향。결과：통과대자행설계합성사이동유사물진행사선，발현A59대우HCT-116구유교강적증식억제작용（IC50=7.6μmol/L）。통과FCM법검측발현A59시통과유도HCT-116세포발생조망병억제세포주기발휘기생물활성。통과Western blot법검측발현A59제량의뢰성격활내질망응격신호통로중적관건단백CHOP，병통과siRNA기인침묵기술진일보학증A59시통과자격CHOP적과표체발휘촉진세포조망적작용。결론：사이동유사물A59통과격활내질망응격신호통도중적관건단백CHOP래발휘대CHT116적증식억제작용。
Objective: To further explore its anti-tumor activity and mechanisms through investigation of the signal path of novel chalcone analogue-A59.Methods: Synthesize the chalcone analogues and further test their cytotoxicity on colon cancer cell line (HCT-116) through MTT method, and pick out the one own bet-ter biological activity named A59. The ability of A59 on inducing HCT-116’s apoptosis and inhibiting its cell cycle were assayed using FCM method. The effect of A59 on ATF4 and CHOP expression in HCT-116 were as-sayed with Western blot. After knockdown CHOP by siRNA technology, the inlfuence of A59 on cell’s apoptosis through immunolfuorescence (IF) and FCM was further investigated.Results:A59 showed inhibitory effects on the proliferation of HCT-116 (IC50=7.6 μm). This novel chalcone analogue exerted its biological activity to induce HCT-116’s apoptosis and inhibit its cell cycle through FCM testing, and the Western blot assay indicated that A59 showed a dose-dependent activation of key protein CHOP of endocytoplasmic reticulum stress (ERS) signal path. and the siRNA technology further conifrmed A59 induce apoptosis by stimulating the CHOP over-expression.Conclusion: Novel chalcone analogue-A59 shows inhibitory effects on the proliferation of HCT-116 by activating the ERS signal path’s key protein CHOP. Clariifes its mechanisms and shows the signiifcance on the new anti-colon cancer drug research and development.