Ma Jian-Xia, Sun Yun-Liang, Yu Yang, Zhang Jian, Wu Hong-Yu, Yu Xiao-Feng
Department of Gastroenterology, Huadong Hospital of Fudan University Shanghai 200338, China.
Department of Gastroenterology, The First People's Hospital of Lianyungang Lianyungang 222061, Jiangsu, China.
Am J Transl Res. 2019 Jun 15;11(6):3750-3760. eCollection 2019.
This study aimed to investigate roles of Toll-like receptor 4 (TLR4)/nuclear factor (NF)-κB signaling in triptolide (TPL)-induced sensitivity of pancreatic cancer cells to gemcitabine (GEM).
, pancreatic cancer PANC-1 cells were treated with lipopolysaccharide (LPS) to activate TLR4, TLR4-siRNA, GEM alone, or GEM plus TPL. , nude mice bearing PANC-1 cell xenografts were treated with GEM, TPL, or both. Cell proliferation was detected by MTT assay and Ki-67 staining. Apoptosis was assessed by flow cytometry and TUNEL assay. A double luciferase reporter gene was used to detect NF-κB activity.
The sensitivity of PANC-1 cells to GEM was reduced by LPS but enhanced by TLR4-siRNA. TPL inhibited expression of TLR4/NF-κB signaling components, which was reversed by LPS. The TPL+GEM group showed more apoptosis than the LPS+TPL+GEM group. Moreover, the activity of NF-κB and the expression of TLR4, p-p65 Survivin, CyclinD1 and Bcl-2 in the TPL+GEM group were lower than in the LPS+TPL+GEM group, whereas Bax expression was higher. The volume of transplanted tumors in the TPL+GEM group was lower than that in the TPL or GEM group. Phospho-p65, Survivin, CyclinD1 and Bcl-2 expression in transplanted tumors was lower in TPL+GEM group than in either single drug group. The Ki-67 staining score of the TPL+GEM group was lower and tumor cells apoptosis rate was increased when compared with TPL or GEM alone.
TPL enhances the sensitivity of pancreatic cancer PANC-1 cells to GEM by inhibiting TLR4/NF-κB signaling.
本研究旨在探讨Toll样受体4（TLR4）/核因子（NF）-κB信号通路在雷公藤甲素（TPL）诱导胰腺癌细胞对吉西他滨（GEM）敏感性中的作用。
用脂多糖（LPS）处理胰腺癌PANC-1细胞以激活TLR4，单独使用TLR4-siRNA、GEM，或GEM加TPL。用GEM、TPL或两者处理携带PANC-1细胞异种移植瘤的裸鼠。通过MTT法和Ki-67染色检测细胞增殖。通过流式细胞术和TUNEL法评估细胞凋亡。使用双荧光素酶报告基因检测NF-κB活性。
LPS降低了PANC-1细胞对GEM的敏感性，但TLR4-siRNA增强了其敏感性。TPL抑制TLR4/NF-κB信号通路成分的表达，LPS可逆转这一作用。TPL+GEM组比LPS+TPL+GEM组显示出更多的细胞凋亡。此外，TPL+GEM组中NF-κB的活性以及TLR4、p-p65、Survivin、细胞周期蛋白D1和Bcl-2的表达低于LPS+TPL+GEM组，而Bax表达更高。TPL+GEM组移植瘤的体积低于TPL组或GEM组。TPL+GEM组移植瘤中磷酸化p65、Survivin、细胞周期蛋白D1和Bcl-2的表达低于单一药物组。与单独使用TPL或GEM相比，TPL+GEM组的Ki-67染色评分更低，肿瘤细胞凋亡率增加。
TPL通过抑制TLR4/NF-κB信号通路增强胰腺癌PANC-1细胞对GEM的敏感性。
