Hedgehog signaling pathway inhibitor reverts acquired chemoresistance in human pancreatic cancer SW1990 cell line
10.3760/cma.j.issn.1674-1935.2012.03.013
- VernacularTitle:Hedgehog通路抑制剂逆转人胰腺癌SW1990细胞获得性耐药的实验研究
- Author:
Jie YAO
;
Yong AN
;
Jianjun QIAN
;
Dousheng BAI
;
Zekuan XU
;
Yi MIAO
- Publication Type:Journal Article
- Keywords:
Pancreatic neoplasms;
Tumor stem cells;
Hedgehog signaling pathway;
Gemcitabine;
Acquired chemoresistance
- From:
Chinese Journal of Pancreatology
2012;12(3):184-188
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the expression levels of hedgehog signaling pathway members (SMO,Gli1,ABCB1,ABCG2) in gemcitabine resistant pancreatic cancer cells and to study the effect of hedgehog signaling pathway inhibitor on acquired chemoresistance.Methods Human pancreatic gemcitabineresistant SW1990 cell line (SW1990/GZ) was established.Hedgehog inhibitor,cyclopamine,was used to treat resistant cells.Different expression levels of SMO,Gli1 and ABCB1,ABCG2 mRNA and proteins,and the percentage of CD44 + CD24 + and CD133 + cells were compared by quantitative RT-PCR and Western blot before and after cyclopamine treatment,respectively.Were treated with 100 μmol/L of gemcitabine SW1990/GZ cells after cyclopamine treatment,and cell apoptosis was determined by Annexin V/PI method.Results The percentage of CD44+ CD24+ and CD133+ cells in SW1990/GZ cells were increased from (29.45 ±1.99)％,(59.37 ± 1.69)％ in parental cells to (93.16 ±2.46)％,(95.88 ± 1.47)％ (P ＜0.05),respectively.The mRNA expression levels of ABCB1,ABCG2,and Gli1 were increased (274.90 ±31.44),(3.48 ±0.33),(4.15 ±0.42) folds,compared with that of parental cells.The mRNA expression of SMO was also detected in resistant cells.The results of protein expression were consisted with that of mRNA expression.After treatment of 2 μmol/L cyclopamine for 1 week,the percentage of CD44 + CD24 + and CD133 +in SW1990/GZ cells decreased to (36.68 ±2.44)％ and (62.76 ± 1.28)％ (P ＜0.05),respectively.After treatment with 2,5μmol/L of cyclopamine,SW1990/GZ cells were re-exposed to 100 μ mol/L of gemcitabine,and then the cell apoptosis and death rates were (53.68 ± 5.24 )％ and (69.99 ± 3.16 )％,which were significantly higher than those in control group ( 4.55 ± 0.87 ) ％ ( P ＜ 0.01 ).Conclusions G emcitabine-resistant pancreatic cancer SW1990 cells are enriched with cancer stem cells and highly express hedgehog member SMO and Glil.Inhibition of hedgehog signaling pathway by cyclopamine can reduce the proportion of cancer stem cell in gemcitabine-resistant cells,down-regulate SMO and Glil expression levels,partly restore the sensitivity to gemcitabine and reverts the acquired drug resistance.
