摘要
目的探讨青蒿琥酯(ART)能否增强顺铂(CDDP)对人结肠癌LOVO细胞的杀伤作用及可能机制。方法通过噻唑蓝法(MTT)计算ART和CDDP对LOVO细胞的IC50值后,分为对照组(等体积PBS)、ART组(10 mol/L)、CDDP组(1 mol/L)和ART(10 mol/L)+CDDP(1 mol/L)联合处理组,免疫荧光检测LOVO细胞β-catenin的表达,荧光素酶报告基因检测TCF-1活性,MTT法及克隆形成实验检测细胞的增殖情况;流式细胞术、比色法和免疫印迹检测细胞凋亡情况。结果 CDDP(1μmol/L)作用48 h后,LOVO细胞β-catenin表达升高且入核,且核内TCF-1活性增强。该现象可被联合使用的ART(10μmol/L)所逆转,同时ART可增强CDDP对LOVO细胞的生长抑制和凋亡诱导。与CDDP单独作用相比,联合使用ART后细胞活力由(89.74±12.13)%降至(47.56±4.15)%,凋亡率(9.25±2.16)%升高至(37.64±9.06)%,Caspase3活性由(130±28)%升高至(310±52)%,差异有统计学意义(P<0.01)。结论 ART能够增强CDDP对LOVO细胞的杀伤作用,其机制可能与抑制Wnt/β-catenin通路及诱导细胞凋亡有关。
Objective To investigate whether Artesunate （ART） could enhance the killing effect of Cisplatin （CDDP） on human colon cancer cell line LOVO and the potential mechanism. Methods The IC50 values of ART and CDDP against LOVO cells were calculated by Methyl thiazolyl tetrazolium （MTY） method, and LOVO cells were divided into blank control group, ART（lOmol/L） group, CDDP （lmol/L） group and ART （10mol/L） ±CDDP （lmol/L） combination group. The expression of β-catenin in LOVO ceils was detected by immunofluorescence （IF）, and the activity of TCF-1 was measured with the dual-luciferase reporter assay system. The cell proliferation was detected by MTY and colony formation assay, and the apoptosis was detected by flow cytometry, colorimetric assay and Western blot analysis. Results The results showed that β- catenin expression increased and accumulated in the nucleus, and the activity of TCF- 1 in the nucleus was enhanced after 48 h treatment with lmol/L CDDP. The results above indicated that the Wnt/β-catenin pathway was activated, However, the results above could be reversed by combined treatment with ART （10mol/L）. Moreover, ART could enhance the growth inhibition and apoptosis induced by CDDP. Compared with the treatment of CDDP alone, the cell viability reduced from （89.74±12.13）% to （47.56±4.15）% （P〈0.01）, the apoptosis rate raised from （9.25±2.16） % to （37.64±9.06） % （P〈0.01）, and the caspase3 activity raised from （130±28） % to （ 310 ±52） % （P〈0.01） after the combination treatment. Conclusions The results of this study conclude that ART could enhance the killing effect of CDDP on LOVO ceils, and the potential mechanism may be related to the apoptosis induced by the inhibition of Wnt/β-catenin pathway.
作者
魏培
黄洁鑫
张矣桐
李娟
刘安定
王志勇
WEI Pei(Key Laboratory of molecular diagnostics in Guangdong Medical University, Guangzhou, Guangdong, 523808, Chin)
出处
《齐齐哈尔医学院学报》
2017年第15期1743-1746,共4页
Journal of Qiqihar Medical University
基金
国家自然科学基金(81503104)
广东省医学科研基金(A2015328
A2016457
A2016158)
广东医科大学大学生创新项目(No.2016ZZDC001)
