Abstract Pancreatic ductal adenocarcinoma is one of the most aggressive tumors with a microenvironment marked by hypoxia and starvation. Galectin-3 has been evaluated in solid tumors and seems to present both pro/anti-tumor effects. So, this study aims to characterize the expression of Galectin-3 from pancreatic tumor cells and analyze its influence for cell survive and motility in mimetic microenvironment. For this, cell cycle and cell death were accessed through flow cytometry. Characterization of inside and outside Galectin-3 was performed through Real-Time Quantitative Reverse Transcription PCR (qRT-PCR), immunofluorescence, Western blot, and ELISA. Consequences of Galectin-3 extracellular inhibition were investigated using cell death and scratch assays. PANC-1 showed increased Galectin-3 mRNA expression when cultivated in hypoxia for 24 and 48 h. After 24 h in simultaneously hypoxic/deprived incubation, PANC-1 shows increased Galectin-3 protein and secreted levels. For Mia PaCa-2, cultivation in deprivation was determinant for the increasing in Galectin-3 mRNA expression. When cultivated in simultaneously hypoxic/deprived condition, Mia PaCa-2 also presented increasing for the Galectin-3 secreted levels. Treatment of PANC-1 cells with lactose increased the death rate when cells were incubated simultaneously hypoxic/deprived condition. Therefore, it is possible to conclude that the microenvironmental conditions modulate the Galectin-3 expression on the transcriptional and translational levels for pancreatic cancer cells.

中文翻译：

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Galectin-3 在缺氧和营养缺乏的情况下在胰腺癌细胞中受到调节

摘要 胰腺导管腺癌是最具侵袭性的肿瘤之一，其微环境以缺氧和饥饿为特征。Galectin-3 已在实体瘤中进行评估，似乎同时具有促/抗肿瘤作用。因此，本研究旨在表征胰腺肿瘤细胞中 Galectin-3 的表达，并分析其在模拟微环境中对细胞存活和运动的影响。为此，通过流式细胞术获得细胞周期和细胞死亡。通过实时定量逆转录 PCR (qRT-PCR)、免疫荧光、蛋白质印迹和 ELISA 对 Galectin-3 的内部和外部进行表征。使用细胞死亡和划痕试验研究了 Galectin-3 细胞外抑制的后果。当在缺氧条件下培养 24 和 48 小时时，PANC-1 显示出 Galectin-3 mRNA 表达增加。在同时缺氧/剥夺孵育 24 小时后，PANC-1 显示出 Galectin-3 蛋白和分泌水平增加。对于 Mia PaCa-2，剥夺培养是 Galectin-3 mRNA 表达增加的决定因素。当同时在缺氧/剥夺条件下培养时，Mia PaCa-2 的 Galectin-3 分泌水平也呈增加趋势。当细胞在缺氧/剥夺条件下同时孵育时，用乳糖处理 PANC-1 细胞会增加死亡率。因此，可以得出结论，微环境条件在胰腺癌细胞的转录和翻译水平上调节 Galectin-3 的表达。PANC-1 显示 Galectin-3 蛋白和分泌水平增加。对于 Mia PaCa-2，剥夺培养是 Galectin-3 mRNA 表达增加的决定因素。当同时在缺氧/剥夺条件下培养时，Mia PaCa-2 的 Galectin-3 分泌水平也呈增加趋势。当细胞在缺氧/剥夺条件下同时孵育时，用乳糖处理 PANC-1 细胞会增加死亡率。因此，可以得出结论，微环境条件在胰腺癌细胞的转录和翻译水平上调节 Galectin-3 的表达。PANC-1 显示 Galectin-3 蛋白和分泌水平增加。对于 Mia PaCa-2，剥夺培养是 Galectin-3 mRNA 表达增加的决定因素。当同时在缺氧/剥夺条件下培养时，Mia PaCa-2 的 Galectin-3 分泌水平也呈增加趋势。当细胞在缺氧/剥夺条件下同时孵育时，用乳糖处理 PANC-1 细胞会增加死亡率。因此，可以得出结论，微环境条件在胰腺癌细胞的转录和翻译水平上调节 Galectin-3 的表达。Mia PaCa-2 也表现出 Galectin-3 分泌水平的增加。当细胞在缺氧/剥夺条件下同时孵育时，用乳糖处理 PANC-1 细胞会增加死亡率。因此，可以得出结论，微环境条件在胰腺癌细胞的转录和翻译水平上调节 Galectin-3 的表达。Mia PaCa-2 也表现出 Galectin-3 分泌水平的增加。当细胞在缺氧/剥夺条件下同时孵育时，用乳糖处理 PANC-1 细胞会增加死亡率。因此，可以得出结论，微环境条件在胰腺癌细胞的转录和翻译水平上调节 Galectin-3 的表达。