Cheng Zhuo-xin, Sun Bei, Wang Shuang-jia, Zhou Hao-xin, Jia Guang, Kong Rui, Wang Gang, Jiang Hong-chi
Department of Pancreatic and Biliary Surgery, the First Affiliated Hospital of Harbin Medical University, Harbin 150001, China.
Zhonghua Wai Ke Za Zhi. 2012 May;50(5):446-51.
To investigate the function of nuclear factor (NF)-κB in the epithelial to mesenchymal transition induced by hypoxia in pancreatic cancer cells.
For cultured pancreatic cancer cells (BxPC-3 and Panc-1) under hypoxic and normoxic conditions, the differences in the morphology were observed by optical microscope. The expression of markers of epithelial and mesenchymal phenotypes, E-cadherin, vimentin and N-cadherin, were determined by Western blot. NF-κB P65 activity was measured by electrophoretic mobility shift assay. Invasion and gemcitabine resistance of pancreatic cancer cells were evaluated in matrigel invasion assay and cell counting kit-8 assay. Both molecular and pharmacologic means of inhibiting NF-κB P65 were used in these hypoxic cells and then the above resulting phenotypes were compared with those of the control-treated cells.
After cultured pancreatic cancer cells under hypoxic conditions for 48 h, normoxic cells exhibited a polygonal shape and formed tight clusters of cells, whereas hypoxic cells took on an elongated, fibroblastoid morphology associated with a more highly invasive character and resistance to gemcitabine; hypoxic cells exhibited an suppression of E-cadherin and increase in vimentin and N-cadherin expression. NF-κB P65 activity was elevated in hypoxic cells. On the contrary, on molecular or pharmacologic inhibition of NF-κB P65, hypoxic cells regained expression of E-cadherin, lost expression of N-cadherin, and reversed their highly invasive and drug resistant phenotype.
Pancreatic cancer cells underwent epithelial to mesenchymal transition exposed to hypoxia, exhibited highly invasive and drug resistant phenotype. Inhibition of NF-κB P65 under hypoxic conditions, pancreatic cancer cells regained expression of E-cadherin, lost expression of N-cadherin, and reversed their highly invasive and drug resistant phenotype.
探讨核因子（NF）-κB在胰腺癌细胞缺氧诱导的上皮-间质转化中的作用。
对培养的胰腺癌细胞（BxPC-3和Panc-1）在缺氧和常氧条件下，通过光学显微镜观察细胞形态的差异。采用蛋白质免疫印迹法检测上皮和间质表型标志物E-钙黏蛋白、波形蛋白和N-钙黏蛋白的表达。通过电泳迁移率变动分析测定NF-κB P65活性。在基质胶侵袭试验和细胞计数试剂盒-8试验中评估胰腺癌细胞的侵袭和吉西他滨耐药性。在这些缺氧细胞中采用分子和药理学方法抑制NF-κB P65，然后将上述所得表型与对照处理细胞的表型进行比较。
将胰腺癌细胞在缺氧条件下培养48小时后，常氧细胞呈多边形，形成紧密的细胞簇，而缺氧细胞呈现细长的成纤维细胞样形态，具有更高的侵袭性和对吉西他滨的耐药性；缺氧细胞表现出E-钙黏蛋白表达受抑制，波形蛋白和N-钙黏蛋白表达增加。缺氧细胞中NF-κB P65活性升高。相反，在分子或药理学上抑制NF-κB P65后，缺氧细胞恢复E-钙黏蛋白表达，失去N-钙黏蛋白表达，并逆转其高侵袭性和耐药表型。
胰腺癌细胞在缺氧条件下发生上皮-间质转化，表现出高侵袭性和耐药表型。在缺氧条件下抑制NF-κB P65，胰腺癌细胞恢复E-钙黏蛋白表达，失去N-钙黏蛋白表达，并逆转其高侵袭性和耐药表型。
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