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灯盏花素对非小细胞肺癌A549细胞的促凋亡作用

花匠小妙招
2025-04-24 10:19

摘要： 目的: 分析灯盏花素对非小细胞肺癌细胞的促凋亡作用以及对移植瘤生长的影响及初步机制探讨。方法: 检测25、50、100 μmol/L灯盏花素处理后A549细胞凋亡、Caspase-3活性及促凋亡蛋白Bax和抗凋亡蛋白Bcl-2表达的改变,用A549细胞构建裸鼠皮下移植瘤模型,每日腹腔注射10 mg/kg和20 mg/kg灯盏花素,每周观察肿瘤的大小、裸鼠体质量,第21天时处死小鼠,获取瘤体,Western blot检测Bax和Bcl-2的表达,分析Bax/Bcl-2比值变化;免疫组化法检测组织内Caspase-3表达变化,TUNEL染色检测肿瘤组织内细胞凋亡情况。结果: 灯盏花素增加A549细胞凋亡率;酶标仪检测结果显示灯盏花素处理后,Caspase-3活性较对照组显著增加;Western blot结果显示灯盏花素能够显著抑制抗凋亡蛋白Bcl-2表达,增加促凋亡蛋白Bax表达,致Bax/Bcl-2比值显著增加。体内实验中,第14和21天时,肿瘤大小较对照组明显减小;而裸鼠的体质量无明显降低。Western blot检测结果显示灯盏花素能够上调Bax的表达,下调Bcl-2的表达,而Caspase-3的表达明显增加。TUNEL染色结果显示灯盏花素处理后,细胞凋亡比例较对照组明显增加。结论: 灯盏花素能够在体内外诱导A549细胞凋亡,其机制可能是灯盏花素通过上调Bax的表达和抑制Bcl-2的表达,增加Bax/Bcl-2的比值,激活Caspase-3,达到促凋亡作用。

关键词: 灯盏花素, 非小细胞肺癌, Bax, Bcl-2, 凋亡

Abstract: AIM: To analyze the apoptosis promoting effect of breviscapine on NSCLC cells and the growth of transplanted tumor and investigate the mechanism. METHODS: The apoptosis, Caspase-3 activity, Bax and Bcl-2 expression of A549 cells treated with 25, 50 and 100 μmol/L breviscapine were detected. The subcutaneous transplanted tumor model was constructed with A549 cells. The mice were intraperitoneally injected with 10 mg/kg and 20 mg/kg breviscapine every day. The size and weight of the tumor were observed every week. On the 21st day, the mice were killed and the tumor was obtained. The expression of Bax and Bcl-2 was detected by Western blot. Bax/Bcl-2 ratio was analyzed. Caspase-3 expression was detected by immunohistochemistry and apoptosis in the tumor was detected by TUNEL staining. RESULTS: Breviscapine increased the apoptosis rate of A549 cells. The results of enzyme labeling showed that Caspase-3 activity increased significantly after breviscapine treatment compared with the control group. Western blot showed that breviscapine could significantly inhibit the expression of anti-apoptosis protein Bcl-2, increase the expression of pro-apoptosis protein Bax, and increase the ratio of Bax/Bcl-2. In vivo, on the 14th and 21st day, the tumor size of the treatment group was significantly smaller than that of the control group, while the weight of nude mice was not significantly reduced. Western blot showed that breviscapine could upregulate the expression of Bax and down regulate the expression of Bcl-2 in the transplanted tumor, while the expression of Caspase-3 was significantly increased. TUNEL staining showed that the proportion of apoptosis increased significantly compared with the control after breviscapine treatment. CONCLUSION: Breviscapine can induce the apoptosis of A549 cells in vitro and in vivo. The mechanism may be that breviscapine upregulates Bax expression and downregulates Bcl-2 expression, increases Bax/Bcl-2 ratio, activates Caspase-3, resulting in A549 cell apoptosis.

Key words: breviscapine, non-small cell lung cancer, Bax, Bcl-2, apoptosis

中图分类号:

R965.2

R734