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目的:探讨光敏化促进姜黄素诱导人胃癌MGC-803细胞凋亡及其机制。方法:用MTT法检测光敏化姜黄素对胃癌MGC-803细胞株的增殖抑制率,Hoechst 33258荧光染色观察细胞核形态的变化,流式细胞术检测细胞的凋亡率、线粒体膜电位、细胞内活性氧和Ca2+;比色法检测caspase-3、8和9酶活性;Western blotting分析细胞色素C、Bcl-2、Bax和热休克蛋白70(HSP70)水平。结果:单纯姜黄素(5.0μmol/L)对MGC-803细胞增殖抑制率为(29.74±2.30)%,在光学显微镜下可见部分凋亡细胞,凋亡率为(12.54±1.75)%。而光敏化姜黄素组细胞增殖抑制率则为(44.93±3.61)%,在光学显微镜下能见明显细胞核形态改变,染色质凝集,凋亡小体形成,凋亡率为(26.58±2.67)%,细胞周期主要阻滞于G0/G1期。光敏化姜黄素显著降低线粒体膜电位,显著增加细胞色素C、细胞内活性氧和Ca2+以及caspase-3、8和9酶活性,与单纯姜黄素组比较,差异显著(P<0.01)。Western blotting结果显示光敏化姜黄素同时显著抑制Bcl-2和HSP70蛋白表达水平。结论:光敏化姜黄素通过Bcl-2和线粒体途径增强其诱导胃癌MGC-803细胞凋亡的作用。
Objective: To investigate the mechanism of photosensitization promoting apoptosis of gastric cancer cell line MGC-803 induced by curcumin. Methods: MTT assay was used to detect the inhibitory effect of photosensitized curcumin on gastric cancer cell line MGC-803. Hoechst 33258 staining was used to observe the change of cell nuclear morphology. Flow cytometry was used to detect the apoptosis rate, mitochondrial membrane potential and intracellular activity Oxygen and Ca2 +. The activities of caspase-3, 8 and 9 were detected by colorimetric assay. The levels of cytochrome C, Bcl-2, Bax and HSP70 were analyzed by Western blotting. Results: The inhibitory rate of curcumin (5.0μmol / L) on proliferation of MGC-803 cells was (29.74 ± 2.30)%. Some apoptotic cells were observed under light microscope. The apoptosis rate was (12.54 ± 1.75)%. (44.93 ± 3.61)%, the apoptotic rate was (26.58 ± 2.67)% in the light-sensitive curcumin group (44.93 ± 3.61)%. The morphological changes of chromatin, chromatin condensation and apoptotic body formation were observed under light microscope. , The main cell cycle arrest in the G0 / G1 phase. Photo-sensitized curcumin significantly decreased mitochondrial membrane potential and significantly increased cytochrome C, intracellular reactive oxygen species, Ca2 + and caspase-3, 8 and 9 enzyme activities compared with curcumin alone group (P <0.01). Western blotting results showed that photosensitized curcumin significantly inhibited the expression of Bcl-2 and HSP70 at the same time. Conclusion: The photosensitized curcumin enhances the apoptosis of gastric cancer MGC-803 cells induced by Bcl-2 and mitochondria.