Liu Xinlu, Weng Yejing, Liu Peng, Sui Zhigang, Zhou Lei, Huang Yinpeng, Zhang Lihua, Zhang Yukui, Tan Xiaodong
First Department of General Surgery, Shengjing Hospital, China Medical University, Shenyang, China.
CAS Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Science, Dalian 116023, China.
Onco Targets Ther. 2018 Jun 6;11:3345-3357. doi: 10.2147/OTT.S162470. eCollection 2018.
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive gastrointestinal cancer characterized by an extremely low survival rate because of early metastasis. Identifying satisfactory therapeutic targets associated with metastasis is crucial to improve the treatment effect of PDAC.
In this research, we used stable isotope labeling by amino acids in cell culture, 1-dodecyl-3-methylimidazolium chloride-assisted sample preparation method preparing protein sample and nano-reversed-phase liquid chromatography-mass spectrometry/mass spectrometry analysis to perform the comparative proteomics of two homologous hamster pancreatic cancer cell lines that are different in metastatic ability: PC-1.0 (highly metastatic) and PC-1 (weakly metastatic). Verifications are through immunohistochemistry on clinical human PDAC pathologic tissues as well as by Western blot of human pancreatic cancer cell lines. siRNA silencing methods were used to study the effect of molecules on invasion and metastasis of pancreatic cancer cell lines.
Bioinformatic analysis indicated that a total of 141 differentially expressed proteins (82 upregulated and 59 downregulated in PC-1.0 cells) were identified showing obviously differential expression (>1.5-fold change). These differentially expressed proteins were involved in a number of different biologic functions, metabolic pathways, and pathophysiologic processes. Phosphoglycerate mutase 1 (PGAM1) and HSPE1 are the top two upregulated proteins, and PDIA3 and CALR are the top two downregulated proteins in PC-1.0 cells compared to PC-1 cells. PGAM1 and HSPE1 showed higher expressions in PDAC tissue with clinical metastasis and highly metastatic pancreatic cancer cell lines PC-1.0 and Aspc-1. PDIA3 and CALR showed higher expressions in weakly metastatic pancreatic cancer cell lines PC-1 and Capan-2. The Western blot results were consistent with the MS quantification data. Silencing PGAM1 was found to decrease the migration and invasion of pancreatic cancer cell lines with statistical significance, especially in highly metastatic PC-1.0 and Aspc-1 cell lines.
These data indicated that PGAM1 may be a potential therapeutic target for PDAC metastasis.
胰腺导管腺癌（PDAC）是一种侵袭性胃肠道癌症，因其早期转移导致生存率极低。确定与转移相关的满意治疗靶点对于提高PDAC的治疗效果至关重要。
在本研究中，我们采用细胞培养中氨基酸稳定同位素标记、1-十二烷基-3-甲基咪唑氯盐辅助样品制备方法制备蛋白质样品，并通过纳米反相液相色谱-质谱/质谱分析，对两种转移能力不同的同源仓鼠胰腺癌细胞系进行比较蛋白质组学研究：PC-1.0（高转移性）和PC-1（低转移性）。通过对临床人PDAC病理组织进行免疫组织化学以及对人胰腺癌细胞系进行蛋白质免疫印迹法进行验证。使用小干扰RNA（siRNA）沉默方法研究分子对胰腺癌细胞系侵袭和转移的影响。
生物信息学分析表明，共鉴定出141种差异表达蛋白（PC-1.0细胞中82种上调，59种下调），其表达差异明显（变化倍数>1.5倍）。这些差异表达蛋白涉及多种不同的生物学功能、代谢途径和病理生理过程。与PC-1细胞相比，磷酸甘油酸变位酶1（PGAM1）和热休克蛋白家族E成员1（HSPE1）是PC-1.0细胞中上调程度最高的两种蛋白，而蛋白二硫键异构酶A3（PDIA3）和钙网蛋白（CALR）是下调程度最高的两种蛋白。PGAM1和HSPE1在伴有临床转移的PDAC组织以及高转移性胰腺癌细胞系PC-1.0和AsPC-1中表达较高。PDIA3和CALR在低转移性胰腺癌细胞系PC-1和Capan-2中表达较高。蛋白质免疫印迹结果与质谱定量数据一致。发现沉默PGAM1可降低胰腺癌细胞系的迁移和侵袭能力，具有统计学意义，尤其是在高转移性PC-1.0和AsPC-1细胞系中。
这些数据表明PGAM1可能是PDAC转移的潜在治疗靶点。
