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

Effect of Crude Extract of Clove Alcohol on the Biological Behaviour of PANC-1 Cells at 39?C

摘要: 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。

Abstract: 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.