Objective

To investigate the role of ribosomal associated protein hnRNP D in resistance to gemcitabine (GEM) in pancreatic cancer cells.

Methods

The expressions of hnRNP D in clinical pancreatic cancer tissues were detected by immunohistochemistry. The proliferation of pancreatic cancer cell lines (PANC-1, BXPC-3, SW1990 and ASPC-1) were measured by CCK8 assay. IC50 of each cell line was calculated and compared. The expressions of hnRNP D protein in pancreatic cancer cell lines were detected by Western Blot assay. The change of hnRNP D expression was confirmed by qPCR and Western Blot after the expressions of hnRNP D in panc-1 cells being down-regulated by miRNA. And than the apoptosis rate and cell cycle of PANC-1 cells were detected by flow cytometry, while the expressions of apoptosis-related proteins cleaved caspase3, P-Akt, AKT and P65 were detected by Western Blot.

Results

HnRNP D protein expressed in clinical pancreatic tissues widely. The IC50 of GEM in PANC-1 was the highest while in BXPC-3 was the lowest. And the expression of hnRNP D protein in PANC-1 was the highest while in BXPC-3 was the lowest. After miRNA interfering, the expressions of hnRNP D protein and gene were significantly decreased in PANC-1 cells. The decrease of hnRNP D expression promoted cell apoptosis and inhibited the cell transformation to the S phase in cell cycle. Under the intervention of GEM, cleaved caspase3 expression was significantly increased, while p-Akt, AKT and P65 expression was significantly decreased.

Conclusion

HnRNP D was associated with resistance to GEM in pancreatic cancer cells. Decreasing of hnRNP D expression promoted cell apoptosis induced by GEM.

中文翻译：

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核糖体相关蛋白 hnRNP D 在吉西他滨耐药胰腺癌中的作用

 客观的

探讨核糖体相关蛋白 hnRNP D 在胰腺癌细胞吉西他滨 (GEM) 耐药中的作用。

 方法

采用免疫组化方法检测临床胰腺癌组织中hnRNP D的表达。通过 CCK8 测定测量胰腺癌细胞系（PANC-1、BXPC-3、SW1990 和 ASPC-1）的增殖。计算并比较各细胞系的IC50。采用Western Blot法检测胰腺癌细胞系中hnRNP D蛋白的表达。 miRNA下调panc-1细胞中hnRNP D表达后，通过qPCR和Western Blot证实hnRNP D表达的变化。流式细胞术检测PANC-1细胞的凋亡率和细胞周期，Western Blot检测凋亡相关蛋白cleaved caspase3、P-Akt、AKT、P65的表达。

 结果

HnRNP D蛋白在临床胰腺组织中广泛表达。 GEM在PANC-1中的IC50最高，而在BXPC-3中最低。 hnRNP D蛋白在PANC-1中表达量最高,在BXPC-3中表达量最低。 miRNA干扰后，PANC-1细胞中hnRNP D蛋白和基因的表达显着降低。 hnRNP D表达减少促进细胞凋亡，抑制细胞向细胞周期S期转化。在GEM的干预下，cleaved caspase3表达显着增加，而p-Akt、AKT和P65表达显着降低。

 结论

HnRNP D 与胰腺癌细胞对 GEM 的耐药性相关。 hnRNP D 表达的减少促进了 GEM 诱导的细胞凋亡。