摘要
目的:研究富半胱氨酸61(cysteine-rich 61,Cyr61)在胰腺癌细胞中对吉西他滨化疗耐药性的影响。方法:培养人胰腺癌细胞株MIAPaCa2,分别感染空白对照病毒,表达Cyr61的病毒或shRNA,用吉西他滨(10μg/ml)处理。细胞克隆形成试验检测细胞增殖情况;流式细胞仪检测胰腺癌细胞凋亡;蛋白质印迹法检测凋亡相关蛋白的表达水平。结果:细胞克隆形成试验显示感染后的稳定细胞中,过表达Cyr61能显著增加细胞对吉西他滨诱导的增殖抑制的耐药性(P<0.05);流式细胞仪检测结果显示,吉西他滨处理后,过表达Cyr61组中胰腺癌细胞凋亡率明显低于对照组(P<0.05);相反,下调内源性的Cyr61减弱了胰腺癌细胞对于吉西他滨诱导的增殖抑制的耐药性(P<0.05),其凋亡率明显高于对照组(P<0.05)。蛋白质印迹结果显示,过表达Cyr61的细胞中,抗凋亡蛋白Bcl-2的表达增加,同时促凋亡蛋白Caspase-2,8,及Cleaved PARP的表达水平降低;下调内源性Cyr61后,凋亡蛋白Bcl-2的表达降低,同时促凋亡蛋白Caspase-2,8,及Cleaved PARP的表达水平升高。结论:Cyr61可以通过调控凋亡相关蛋白的表达水平,从而增强胰腺癌细胞对吉西他滨的化疗耐药性。
Objective： To investigate the effect of cysteine-rich 61（Cyr61） on the chemoresistance of human pancreatic cancer cells induced by gemeitabine.Methods： Human pancreatic cancer cells MIA PaCa2 were cultured and infected with empty vector,vector expression Cyr61 or shRNA.The ability of cell proliferation was analyzed by clone formation assay;the apoptosis was evaluated by the flow cytometry analysis stained with propidium iodide.The expression of the apoptosis related proteins were examined by Western blot assay.Results： As shown in clone formation assay,Cyr61 enhanced the antiproliferation capability of MIA PaCa2 to gemcitabine（P0.05）.Compared with control group,the MIA PaCa2 forced expression of Cyr61 had less apoptosis rate（P0.05）.On the contrary,knockdown the endogenous Cyr61 inhibited the antiproliferation capability to gemcitabine as well as enhanced the apoptosis rate（P0.05）.Western blot shown that there were increased Bcl-2 expression and decreased caspase-2,8 and cleaved PARP expression in Cyr61 forced expression group compared with control groups,while there were decreased Bcl-2 expression and increased caspase-2,8 and cleaved PARP expression in MIA PaCa2/Cyr61i group.Conclusions： Cyr61 endowed the chemoresistance ability of pancreatic cancer cell to gemcitabine through inhibiting the pathway.
出处
《江苏大学学报（医学版）》
CAS
2011年第4期308-312,共5页
Journal of Jiangsu University：Medicine Edition
基金
镇江市社会发展基金资助项目(SH2009013)
