Study on Inhibitory Effects and Mechanism of Bruceanine D Combined with Taxol on the Proliferation of Human Pancreatic Cancer Capan- 2 Cells
- VernacularTitle:鸦胆子素D联合紫杉醇对胰腺癌Capan-2细胞增殖的抑制作用及机制研究
- Author:
Yuyu HUANG
1
,
2
;
Mingjun RAO
1
,
2
;
Biqin TAN
1
,
3
;
Huiming WANG
2
;
Nengming LIN
1
,
2
Author Information
1. No. 4 Clinical Medical School,Zhejiang University of TCM,Hangzhou 310006,China
2. College of Pharmacology,Zhejiang University of TCM,Hangzhou 310053,China
3. Dept. of Clinical Pharmacology,the Affiliated Hangzhou First People’s Hospital,Zhejiang University School of Medicine,Hangzhou 310006,China
- Publication Type:Journal Article
- Keywords:
Brucerin D;
Taxol;
Human pancreatic cancer;
Capan-2 cells;
Proliferation;
Apoptosis;
Caspase;
JNK
- From:
China Pharmacy
2019;30(6):789-795
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE: To investigate the inhibitory effect and potential mechanism of Brucein D (BD) combined with Taxol on the proliferation of human pancreatic cancer Capan-2 cells. METHODS: Using Capan-2 cells as object, the proliferations after treated with BD (5, 10, 15, 20 μmol/L), Taxol (10, 20, 30, 40 nmol/L) and BD+Taxol (5 μmol/L+10 nmol/L, 10 μmol/L+20 nmol/L, 15 μmol/L+30 nmol/L, 20 μmol/L+40 nmol/L) for 48 h were determined by sulfonyl rhodamine B method. Survival rate of cells and combination index (CI) were calculated. The clone formation assay was performed to detect the formation of clonal colonies after treated with BD (20 μmol/L,hereinafter), Taxol (40 nmol/L,hereinafter)、BD+Taxol (20 μmol/L+40 nmol/L,hereinafter) for 24 h. The rate of clone formation was calculated. DAPI method was used to observe the apoptosis of cells after treated with BD, Taxol and BD+Taxol for 24 h. Western blotting was used to detect the expression of apoptosis-related protein (Bcl-2, PARP, Caspase-3, Cleaved-caspase-3) after treated by BD, Taxol, BD+Taxol for 48 h and the expression of JNK and p-JNK after treated by BD, Taxol, BD+Taxol for 4, 6, 12 h. RESULTS: After treated with 10, 15 and 20 μmol/L BD, 20, 30 and 40 nmol/L Taxol or two-drug combination for 48 h, survival rates of cells were decreased significantly; the survival rate of drug combination group was significantly lower than the same dose of BD group and Taxol group (P<0.05). CI values of drug combination groups (BD 5 μmol/L+Taxol 10 nmol/L, BD 10 μmol/L+Taxol 20 nmol/L, BD 15 μmol/L+Taxol 30 nmol/L, BD 20 μmol/L+Taxol 40 nmol/L) were 0.63±0.04, 0.68±0.08, 0.89±0.12 and 0.84±0.05. After treated with 20 μmol/L BD, 40 nmol/L Taxol and two-drug combination, the formation of clonal colonies was decreased with different degrees of chromatin concentration and nuclear shrinkage; the rate of clone formation (24 h), the expression of Bcl-2 (48 h), PARP (48 h), Caspase-3 (48 h) and JNK (4, 6 h, except for Taxol group) were decreased significantly, while the relative expression of Cleaved-caspase-3 (48 h) and p-JNK (4, 6, 12 h) were increased significantly. Those of BD+Taxol group were significantly better than those of BD group and Taxol group [except for JNK (4, 6, 12 h), p-JNK (4 h)] (P<0.05 or P<0.01). CONCLUSIONS: Both BD and Taxol can inhibit the proliferation and promote apoptosis of human pancreatic cancer Capan-2 cells, and the combination have a certain synergistic effect, which is better than any single drug. It may be associated with activating Caspase pathway and JNK phosphorylation.