Feng Jiake, Ma Tieliang, Ge Zhijun, Lin Jie, Ding Weiliang, Chen Hong, Zhu Wenjiao, Zhou Sujun, Tan Yongfei
Department of General Surgery, The Affiliated Yixing Hospital of Jiangsu University, Yixing, Jiangsu 214200, P.R. China.
Central Laboratory, The Affiliated Yixing Hospital of Jiangsu University, Yixing, Jiangsu 214200, P.R. China.
Mol Med Rep. 2015 Mar;11(3):2111-7. doi: 10.3892/mmr.2014.2990. Epub 2014 Nov 20.
The aim of the current study was to investigate the effect of the PKM2 gene on the proliferation, invasion, migration and apoptosis of Panc‑1 and Sw1990 pancreatic cancer cells via its interaction with the mitogen‑activated protein kinases (MAPKs) signaling pathways. The expression levels of PKM2 protein in pancreatic cancer cells and the corresponding normal tissues was determined with western blot analysis. Immunohistochemical analysis of PKM2 expression was carried out in paraffin‑embedded sections of pancreatic cancer tissue. Two human pancreatic cancer cell lines were cultured in vitro, and a small interfering RNA (siRNA) was designed for the PKM2 gene and transfected into the cells. Cell proliferation was measured via an MTT assay, cell migration and invasion was measured via Transwell® chambers, and the effect of PKM2 on apoptosis was detected from B‑cell lymphoma 2 (Bcl‑2) and Bcl‑2‑associated X protein expression levels. Protein expression levels of the MAPK pathway proteins extracellular signal‑regulated kinase 1/2 (ERK1/2), p38 and c‑Jun N‑terminal kinase (JNK) and their phosphorylated forms were measured via western blot analysis. The expression level of PKM2 was significantly upregulated in the pancreatic cancer tissue compared with that of the corresponding normal tissue. Downregulation of PKM2 expression reduced the proliferation, migration and invasion of pancreatic cancer cell lines, while increasing the levels of apoptosis. Additionally, the expression levels of the phosphorylated‑(p‑)ERK1/2 and p‑p38 of the MAPK pathway in the PKM2 siRNA groups were markedly downregulated compared with those of the controls; however, the expression levels of ERK1/2, p38, JNK, p‑p38 and p‑JNK had no significantly changes compared with those of the control groups. In summary, the PKM2 gene has an important role in the proliferation, invasion, migration and apoptosis of Panc‑1 and Sw1990 pancreatic cancer cells, which may be associated with the expression of ERK1/2 and p38 of the MAPK signaling cascade.
本研究旨在通过PKM2基因与丝裂原活化蛋白激酶（MAPKs）信号通路的相互作用，探讨其对Panc-1和Sw1990胰腺癌细胞增殖、侵袭、迁移及凋亡的影响。采用蛋白质免疫印迹分析检测胰腺癌细胞及相应正常组织中PKM2蛋白的表达水平。对胰腺癌组织石蜡包埋切片进行PKM2表达的免疫组织化学分析。体外培养两个人胰腺癌细胞系，设计针对PKM2基因的小干扰RNA（siRNA）并转染至细胞中。通过MTT法检测细胞增殖，通过Transwell®小室检测细胞迁移和侵袭，并从B细胞淋巴瘤2（Bcl-2）和Bcl-2相关X蛋白表达水平检测PKM2对凋亡的影响。通过蛋白质免疫印迹分析检测MAPK通路蛋白细胞外信号调节激酶1/2（ERK1/2）、p38和c-Jun氨基末端激酶（JNK）及其磷酸化形式的蛋白表达水平。与相应正常组织相比，胰腺癌组织中PKM2的表达水平显著上调。PKM2表达下调可降低胰腺癌细胞系的增殖、迁移和侵袭，同时增加凋亡水平。此外，与对照组相比，PKM2 siRNA组中MAPK通路的磷酸化-（p-）ERK1/2和p-p38的表达水平明显下调；然而，与对照组相比，ERK1/2、p38、JNK、p-p38和p-JNK的表达水平无显著变化。综上所述，PKM2基因在Panc-1和Sw1990胰腺癌细胞的增殖、侵袭、迁移及凋亡中起重要作用，这可能与MAPK信号级联中ERK1/2和p38的表达有关。
