Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Jan-Dec:19:1533033819895494.
doi: 10.1177/1533033819895494.

Upregulation of DAB2IP Inhibits Ras Activity and Tumorigenesis in Human Pancreatic Cancer Cells

Yifan Duan  1   2 , Xiaoyu Yin  3 , Xiaorong Lai  4 , Chao Liu  5 , Wenjing Nie  6 , Dongfeng Li  7 , Zijun Xie  2   8 , Zijun Li  2   9 , Fan Meng  2   10
Affiliations

Upregulation of DAB2IP Inhibits Ras Activity and Tumorigenesis in Human Pancreatic Cancer Cells

Yifan Duan et al. Technol Cancer Res Treat. 2020 Jan-Dec.

Abstract

KRAS mutation-induced Ras activation plays an important role in the pathogenesis of pancreatic cancer, but the role of wild-type Ras and Ras GTPase-activating proteins remains unclear. The present study was designed to determine the expression spectra of Ras GTPase-activating proteins genes in pancreatic cancer cells, and the role of DAB2IP, a Ras GTPase-activating proteins gene, in the development and progression of pancreatic cancer. Following the analyses of the expression profiles of 16 Ras GTPase-activating proteins in 6 pancreatic cancer cell lines including Bxpc-3 (with wild-type KRAS), Capan-2, Sw1990, Aspc-1, CFPAC-1, and Panc-1 (with mutant KRAS) and 1 normal human pancreatic ductal epithelial cell line, H6C7, the expression of DAB2IP messenger RNA was further analyzed by quantitative real-time polymerase chain reaction. The role of DAB2IP in pancreatic cancer was further investigated in vitro and in vivo by upregulating DAB2IP in Bxpc-3 cells through transfection of DAB2IP into Bxpc-3 cells with recombinant lentivirus. The DAB2IP expression in pancreatic cancer cells and tissues with wild-type KRAS was significantly lower than that in cells and tissues with mutant KRAS (P < .05). In Bxpc-3 cells with wild-type KRAS, overexpression of DAB2IP decreased the expression of P-AKT and P-ERK and the Ras activity; increased the expression of P-JNK and caspase 3; inhibited cell proliferation, invasiveness, and migration; and increased the cell sensitivity to cetuximab. Overexpression of DAB2IP inhibited tumor progression in a mouse model. In conclusion, DAB2IP downregulates Ras activity in wild-type pancreatic cancer cells. Overexpression of DAB2IP decreases the Ras activity, inhibits cell proliferation, and increases sensitivity to cetuximab in wild-type pancreatic cancer cells. In conclusion, DAB2IP may serve as a potential molecular therapeutic target for the treatment of pancreatic cancer.

Keywords: Bxpc-3 cells; DAB2IP; Ras activity; RasGAP; pancreatic cancer; tumor suppressors.

PubMed Disclaimer

Conflict of interest statement

Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Figures

Figure 1.
Figure 1.
The messenger RNA (mRNA) expression levels of 16 Ras GTPase-activating proteins (GAPs) in 6 pancreatic cancer cell lines and a normal pancreatic ductal cell line. The RasGAPs superfamily includes 16 members: RASAL3, RASA2, RASA3, IQGAP2, IQGAP3, SYNGAP1, GAPVD1, IQGAP1, ARHGAP5, RASAL2, RASA4, G3BP1, NF1, DAB2IP, RASAL1, and RASA1. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to analyze the RasGAPs mRNA levels in pancreatic cancer cells (expressing wild-type KRAS: Bxpc-3; expressing mutant KRAS: Capan-2, Sw1990, CFPAC-1, Aspc-1, Panc-1) and normal H6C7 cells. # P < .05, pancreatic cancer cells versus H6C7 cells; *P < .05, pancreatic cancer cells with wild-type KRAS gene versus pancreatic cancer cells with a mutant KRAS gene.
Figure 2.
Figure 2.
The expression levels of DAB2IP protein in pancreatic cancer tissues and controls, as analyzed by immunohistochemistry. (A) positive control (breast cancer); (B) negative control (pancreatic cancer, phosphate-buffered saline [PBS] was substituted for the primary antibody); (C) normal pancreatic tissue; (D) pancreatic cancer tissue with wild-type KRAS; (E) pancreatic cancer tissue with mutant KRAS; and (F) adjacent tissue. Magnification: ×400.
Figure 3.
Figure 3.
Validation of lentivirus transfection efficiency on Bxpc-3 cells and role of DAB2IP in the Ras signaling pathway. (A and B) The DAB2IP messenger RNA (mRNA) and protein expression levels were increased in wild-type KRAS pancreatic cancer cells after lentivirus transfection (P < .05) as assessed by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting, respectively. (C and D), Overexpression of DAB2IP decreased the expression of P-AKT, P-ERK, and Bcl-2, coupled with increased expression of P-JNK and caspase 3 (P < .05), as assessed by Western blotting. (E) Overexpression of DAB2IP decreased the Ras activity (P < .05), as assessed by RAS-GTP pulldown assay; and (F and G) DAB2IP overexpression decreased the proliferation and colony formation of wild-type KRAS pancreatic cancer cells (P < .05), as assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and colony formation assays.
Figure 4.
Figure 4.
Effects of DAB2IP overexpression on cell invasion, migration, apoptosis, and cell cycle. (A and B) Overexpression of DAB2IP weakened the invasion and migration capacity of Bxpc-3 cells (P < .05), as assessed by Transwell cell invasion and wound healing assays. (C and D) Overexpression of DAB2IP increased cell apoptosis and the percentage of cells in the G0/G1 phase (P < .05), as assessed by Annexin V/propidium iodide assay.
Figure 5.
Figure 5.
Effects of DAB2IP overexpression on cell sensitivity to cetuximab in vitro and on tumor growth in vivo. (A) Compared to the control Bxpc3-Vector cells, Bxpc3-DAB2IP + pancreatic cancer cells were more inhibited by cetuximab at various doses (50, 100, and 200 µg/mL). Overexpression of DAB2IP enhanced the sensitivity of the pancreatic cancer cells with wild-type KRAS to cetuximab, compared to the control cells (P < .05), as assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. (B-D) The tumor volume and weight were smaller in Bxpc3-DAB2IP + cetuximab group than in the Bxpc3-Vector + cetuximab group (P < .05), as assessed by nude mouse tumorigenicity assay. B refers to how the tumor volume in the Bxpc3-DAB2IP + cetuximab group was inhibited more than that in the control Bxpc3-Vector + cetuximab group, as well as inhibited more in the Bxpc3-DAB2IP group compared to the Bxpc3-Vector group (P < .05).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Hidalgo M. Pancreatic cancer. N Engl J Med. 2010;362:1605–1617. - PubMed
    1. Rossi ML, Rehman AA, Gondi CS. Therapeutic options for the management of pancreatic cancer. World J Gastroenterol. 2014;20(32):11142–11159. - PMC - PubMed
    1. Di Marco M, Astolfi A, Grassi E, et al. Characterization of pancreatic ductal adenocarcinoma using whole transcriptome sequencing and copy number analysis by single-nucleotide polymorphism array. Mol Med Rep. 2015;12(5):7479–7484. - PubMed
    1. Sano M, Driscoll DR, DeJesus-Monge WE, et al. Activation of WNT/β-catenin signaling enhances pancreatic cancer development and the malignant potential via up-regulation of Cyr61. Neoplasia. 2016;18(12):785–794. - PMC - PubMed
    1. Hennig A, Markwart R, Esparza-Franco MA, et al. Ras activation revisited: role of GEF and GAP systems. Biol Chem. 2015;396(8):831–848. - PubMed

Publication types

MeSH terms

Substances