【摘要】 目的:研究应用顺铂联合姜黄素对诱导胃癌细胞HGC27凋亡的影响及机制。方法:用CCK8法检测单独或联合使用不同剂量顺铂或/和姜黄素对胃癌细胞HGC27增殖的影响;用Hochest33258染色检测联合应用顺铂和姜黄素诱导胃癌细胞HGC27凋亡的发生率。用Western blot检测细胞凋亡相关分子PARP1蛋白剪切体和DNA损伤蛋白p-γH2AX的表达变化。结果:单用顺铂可以呈剂量依赖性地促进HGC27细胞凋亡。5μmol·L-1姜黄素对HGC27细胞增殖无明显作用,10μmol·L-1姜黄素反而轻微促进HGC27细胞增殖。20、40、80μmol·L-1姜黄素对HGC27细胞的增殖抑制率分别为10.97%、15.15%、32.93%。分析联合用药组:5μmol·L-1姜黄素联合顺铂组反而促进HGC27细胞生长;10μmol·L-1姜黄素联合不同剂量顺铂和单用顺铂组比较,组间抑制率没有统计学差异(P>0.05)。20μmol·L-1姜黄素联合4、6μmol·L-1顺铂有明显的促进细胞的凋亡作用,抑制率分别为70.68%、87.30%。Hochest33258染色结果显示,联合用药组细胞凋亡小体和坏死细胞明显增多。Western blot结果显示,在联合用药组HGC27细胞PARP1剪切体蛋白和p-γH2AX蛋白表达明显增加。结论:顺铂联合姜黄素可以促进胃癌细胞HGC27的凋亡,机制是姜黄素可以加重顺铂引起的细胞DNA双链损伤。更多还原

【Abstract】 Objective: To investigate the role of curcumin in cisplatin induced apoptosis of human gastric cancer HGC27 cells. Methods: Cytotoxicity of cisplatin and curcumin were detected by CCK-8assay. The amount of apoptotic cells was detected by hochest 33258 staining assay. The DNA damage biomarker p-γH2AX and cell apoptosis biomarker cleaved-PARP1 were detected by Western blot. Results:Cisplatin induced HGC27 cells death in a dose-dependent manner. The proliferation of HGC27 cells was not affected by curcumin at 5 μmol·L-1, lightly enhanced at 10 μmol·L-1, while inhibited at 20, 40 and 80μmol·L-1for 10.97%, 15.15 % and 32.93 %, respectively. The combination of 5 or 10 μmol·L-1curcumin with different concentration of cisplatin could not induce HGC27 appoptosis effectively(P>0.05). However,significantly increased cell death was observed in HGC27 cells with the combination of 20 μmol·L-1curcumin and 4 or 6 μmol ·L-1cisplatin. Hoechst33258 staining showed that apoptotic cells were significantly increased in the combined group. Western blot results also showed that the levels of DNA damage marker p-γH2AX and apoptosis marker cleaved-PARP1 were significantly increased in the combined group. Conclusion: Curcumin enhances cisplatin induced apoptosis by inhibiting DNA repair in human gastric cancer HGC27 cells.更多还原