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丁香酒精粗提取物在39℃条件下对PANC-1细胞生物学行为的影响
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Abstract:
目的：探究丁香酒精粗提取物在39℃条件下是否会影响胰腺癌细胞的增殖、迁移、侵袭和凋亡。方法：1) CCK8法检测在39℃条件下丁香酒精提取物对胰腺癌细胞增殖活性的影响；2) 平板克隆形成和软琼脂克隆法检测在39℃条件下丁香酒精提取物对胰腺癌细胞生长和增殖的影响；3) 划痕实验检测在39℃条件下丁香酒精提取物对胰腺癌细胞迁移和侵袭能力的影响；4) Hoechst33258染色和流式细胞术检测在39℃条件下丁香酒精提取物对细胞形态的改变及凋亡的影响。结果：CCK8结果表明，丁香酒精粗提取物可以抑制PANC-1细胞的生长和增殖，抑制作用呈浓度依赖性，联合39℃效果更加明显，划痕实验实验结果表明，丁香酒精粗提取物可以抑制PANC-1细胞迁移，联合39℃效果更加明显，但是37℃和39℃对照没有明显变化，显微镜下观察加入丁香酒精粗提取物后，PANC-1细胞出现明显的变化，从规则的菱形变成了细长无边界感变圆形态发生了变化，Hoechst33258实验结果显示丁香酒精粗提取物可以引起PANC-1细胞的细胞核出现亮蓝色、凋亡小体和核碎裂等现象，流式细胞术也验证了丁香酒精粗提取物可以诱导PANC-1细胞的凋亡39℃条件下凋亡细胞数增加。结论：丁香酒精粗提取物可以抑制细胞生长增殖，抑制细胞的迁移侵袭并诱导细胞的凋亡，并联合39℃效果更加明显，从而从不同的机制来发挥抗肿瘤作用。
Objective: To explore whether the crude extract of clove alcohol at 39？C affects the proliferation, migration, invasion and apoptosis of pancreatic cancer cells. Methods: 1) CCK8 assay to detect the effect of crude extract of clove alcohol on the proliferation activity of pancreatic cancer cells at 39？C; 2) plate clone formation and soft agar cloning assay to detect the effect of crude extract of clove alcohol on the growth and proliferation of pancreatic cancer cells at 39？C; 3) scratch assay and Transwell assay to detect the effect of crude extract of clove alcohol on pancreatic cancer cells at 39？C; 4) Hoechst 33258 staining and flow cytometry to detect the effect of clove alcohol extract on cell morphological changes and apoptosis at 39？C. Results: The results of CCK8 showed that the crude extract of clove alcohol could inhibit the growth and proliferation of PANC-1 cells, and the inhibitory effect was concentration-dependent, and the effect was more obvious in combination with 39？C. The results of the scratch test showed that the crude extract of clove alcohol could inhibit the migration of PANC-1 cells, and the effect was more obvious in combination with 39°C, but there was no significant change between 37？C and 39？C control. The results of Hoechst33258 experiment showed that the crude extract of clove alcohol could cause the nucleus of PANC-1 cells to appear bright blue, apoptotic vesicles and nuclear fragmentation. The flow cytometry also verified that the crude extract of clove alcohol could induce apoptosis in PANC-1 cells with an increase in the number of apoptotic cells at 39？C. Conclusion: The crude extract of clove alcohol can inhibit cell growth and proliferation, inhibit cell migration and invasion and induce apoptosis, and the effect is more obvious in combination with 39？C, thus exerting anti-tumor effects from different mechanisms.
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