青藤碱通过调控NF-κB信号通路抑制胰腺癌Capan-1细胞增殖并诱导细胞凋亡

Sinomenine Inhibits Proliferation and Induces Apoptosis of Pancreatic Cancer Capan-1 Cells by Regulating NF-κB Signaling Pathway

目的探讨青藤碱对胰腺癌Capan-1细胞增殖、凋亡的影响及机制。方法CCK8法检测细胞活力,DAPI/TUNEL双染检测细胞凋亡,流式细胞术Annexin V-FITC/PI染色法检测细胞凋亡率,Western blotting检测cleaved caspase-3蛋白、细胞核NF-κB蛋白表达。结果CCK8结果表明青藤碱抑制胰腺癌Capan-1细胞增殖,并具有时间、浓度依赖性;DAPI/TUNEL染色结果表明,在青藤碱作用下,胰腺癌Capan-1细胞凋亡增多;流式细胞术Annexin V-FITC/PI结果表明,在青藤碱作用下,胰腺癌Capan-1细胞凋亡率增高;Westernblotting结果表明,在青藤碱作用下,胰腺癌Capan-1细胞cleavedcaspase-3表达升高,细胞核NF-κB表达降低。TNF-α降低青藤碱对胰腺癌Capan-1细胞增殖抑制作用、凋亡诱导作用和cleavedcaspase-3表达的上调作用。结论青藤碱通过调控NF-κB信号通路抑制胰腺癌Capan-1细胞增殖并诱导细胞凋亡。

OBJECTIVE To investigate the effect and mechanism of sinomenine on apoptosis of pancreatic cancer Capan-1 cells.METHODS CCK8 assay was used to detect cell viability.DAPI/TUNEL staining was used to detect cell apoptosis.Flow cytometry Annexin V-FITC/PI was used to detect cell apoptotic rate.Western blotting was used to detect the expression of cleaved caspase-3 and nuclear NF-κB protein.RESULTS Result of CCK8 assay showed that sinomenine can inhibit the proliferation of Capan-1 cells effectively in a dose and time dependent manner.Result of DAPI/TUNEL staining showed that the apoptosis of Capan-1 cells increased by treated with sinomenine.Flow cytometric analysis staining with Annexin V-FITC/PI showed that the apoptosis rates of Capan-1 cells increased by treated with sinomenine.Result of Western blotting analysis showed that the expression of cleaved caspase-3 increaed and the expression of nuclear NF-κB decreased in Capan-1 cells by treated with sinomenine.TNF-a could reversed the effect of sinomenine on the proliferation and apoptosis of Capan-1 cell.TNF-a could reversed the up-regulated effect of sinomenine on the expression of cleaved caspase-3 in Capan-1 cells.CONCLUSION Sinomenine inhibits proliferation and induces apoptosis of pancreatic cancer Capan-1 cells by regulating the NF-κB signaling pathway.