Cyclopamine reverts acquired chemoresistance and down-regulates cancer stem cell markers in pancreatic cancer cell lines.
Cyclopamine reverts acquired chemoresistance and down-regulates cancer stem cell markers in pancreatic cancer cell lines.
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DOI：
10.4414/smw.2011.13208
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发表时间：
2011-05
影响因子：
2.9
通讯作者：
Jie Yao;Yong An;Jiajia Wie;Zhen-ling Ji;Zipeng Lu;Junli Wu;K. Jiang;Ping Chen;Ze-Kuan Xu;Y. Miao
中科院分区：
文献类型：
--
作者：
Jie Yao;Yong An;Jiajia Wie;Zhen-ling Ji;Zipeng Lu;Junli Wu;K. Jiang;Ping Chen;Ze-Kuan Xu;Y. Miao
Hedgehog（Hh）通路与包括胰腺导管腺癌（PDAC）在内的癌症的发病机制有关。最近的研究表明，Hh在维持癌症干细胞（CSCs）库中起着重要作用。吉西他滨耐药胰腺癌细胞高度表达一些CSC标志物。然而，Hh成员在吉西他滨耐药胰腺癌细胞中的表达水平仍然未知。本研究的目的是验证HH成员，如Shh，Ptc，SMO和Gli-1在吉西他滨耐药的PDAC细胞系中的表达，并探索克服PDAC化疗耐药性的新策略。材料与方法采用实时荧光定量PCR（Q-PCR）和western blot检测HH基因在SW 1990、CFPAC-1细胞和吉西他滨耐药细胞中的相对表达水平。采用流式细胞术、Q-PCR、western blot检测cyclopamine治疗前后肿瘤干细胞标志物及HH成员表达水平的变化。流式细胞仪检测环巴胺作用后细胞凋亡情况。结果吉西他滨耐药细胞高表达CD 44、CD 133及HH成员Shh、SMO、Gli-1，环巴胺处理后，其表达水平明显下调。流式细胞术分析显示环巴胺处理后细胞凋亡增加。结论胰腺癌吉西他滨耐药细胞高表达CSCs标志物和部分HH成员，环巴胺抑制HH是逆转胰腺癌吉西他滨耐药的有效方法。
BACKGROUND The hedgehog (Hh) pathway has been implicated in the pathogenesis of cancer including pancreatic ductal adenocarcinoma (PDAC). Recent studies have suggested that Hh plays an important role in maintaining the cancer stem cell (CSCs) pool. Gemcitabine-resistant pancreatic cancer cells highly express some of the CSCs markers. However, the expression level of Hh members in gemcitabine-resistant pancreatic cancer cells remains unknown. The aim of this study was to verify the expression of HH members, such as Shh, Ptc, SMO and Gli-1 in gemcitabine-resistant PDAC cell lines, and to explore a new strategy to overcome chemoresistance in PDAC. MATERIAL AND METHODS Quantitative real-time RT-PCR (Q-PCR) and western blot were used to evaluate the relative expression level of HH members in SW1990, CFPAC-1 cells and gemcitabine-resistant SW1990, CFPAC-1 cells. The change of cancer stem cell markers and the expression level of HH members before and after cyclopamine treatment was evaluated using flow cytometry and Q-PCR, western blot, respectively. Cell apoptosis after cyclopamine treatment was measured by flow cytometry. RESULTS CD44, CD133 and the expression level of HH members, including Shh, SMO, Gli-1, were found to be highly expressed in gemcitabine-resistant cells, which were significantly down-regulated by cyclopamine treatment. Flow cytometry analysis showed increased cell apoptosis after cyclopamine treatment. CONCLUSION Gemcitabine-resistant pancreatic cancer cells highly express CSCs markers and some of the HH members, and inhibition of HH by cyclopamine is an effective method of reversing gemcitabine resistance in pancreatic cancer.