Original ArticlesKindlin-2 induced by TGF-β signaling promotes pancreatic ductal adenocarcinoma progression through downregulation of transcriptional factor HOXB9
Introduction
Pancreatic ductal adenocarcinoma (PDAC) accounts for 85–90% of pancreatic cancer and is the fourth leading cause of cancer-related deaths with five-year overall survival rates of 5–6% [1]. Pancreatic cancer is characterized by invasion to adjacent organs and metastasis to remote organs at the early stage [2]. Surgery, radiotherapy and chemotherapy have limited effects on pancreatic cancer. The average five-year overall survival rises from 2% in the years 1975–1977 to 6% in the years 2003–2009 for patients with pancreatic cancer [3]. Therefore, there are urgent needs to identify new molecules and pathways that control PDAC progression.
Kindlin-2, encoded by FERMT2 gene, controls integrin activation through binding to the integrin β cytoplasmic tails [4], [5], [6]. Kindlin-2 plays important roles in a variety of malignancies including lung, breast, and high-grade invasive bladder cancers [7]. Recently it was reported that Kindlin-2 promotes invasion of gastric cancer cells through phosphorylation of integrin β1 and β3 cytoplasmic domains [8]. Kindlin-2 is also a new regulator of Wnt signaling and regulates cancer progression in an integrin-binding independent manner [9]. Inhibition of Kindlin-2 suppresses invasion of esophageal squamous carcinoma cells through modulation of the cytoskeleton [10]. In addition, Kindlin-2 was found to express in the stroma of pancreatic cancer and is associated with poor prognosis of the patients [11]. However, little is known about the role of Kindlin-2 in PDAC progression, and how Kindlin-2 regulates migration and invasion of the PDAC cells.
Transforming growth factor-β1 (TGF-β1) is one of the important growth factors regulating tumor invasion and metastasis through epithelial to mesenchymal transition (EMT) [12], [13], [14]. Deregulation of TGF-β1 had been known to contribute to the progression of pancreatic cancer. However, the molecular mechanism was not fully elucidated. Kindlin family member Kindlin-1 was found to be induced by TGF-β1 in human mammary epithelial cells [15]. We recently found that Kindlin-2 physically interacts with both TβRI and Smad3 and promotes the activation of TGF-β/Smad signaling in tubule interstitial fibrosis [16]. However, less is known about how Kindlin-2 participates in the TGF-β signaling pathway during cancer progression.
HOXB9 is a member of the homeobox-containing (HOX) transcriptional factor family that encompasses 39 genes in humans and is classified into four different clusters, including HOX A, B, C and D [17]. In addition to their critical roles in development, increasing evidence demonstrated that HOX family genes are associated with cancer progression [18], [19], [20]. HOXB9 was known to induce angiogenesis, invasion and lung metastasis in breast cancer [21], and being an important prognostic factor for breast cancer [22], [23], [24], [25] as well as for lung cancer patients [26]. On the opposite, enhanced expression of HOXB9 was recently found related to a favorable overall survival in patients with gastric carcinoma and colon adenocarcinoma [27], [28]. These findings suggested that HOXB9 may play a diverse role during cancer progression under various circumstances. However, until now HOXB9 was not linked to pancreatic cancer progression.
In this study, we investigated the regulation between Kindlin-2 and TGF-β signaling or Kindlin-2 and HOXB9 in PDAC progression. Our investigations helped to understand the molecular mechanisms underlying PDAC progression and to identify potential therapeutic targets for PDAC.
Section snippets
Ethics
The Ethics Committee of Peking University Health Science Center has approved the mouse experiments (Permit Number: LA2011-73). PDAC patients' tumor tissues in the present study have been approved by Sino-Japan Friendship Hospital (Permit Number: ZRLW-5). The protocol for handling mice and human tumor specimens was in accordance with the ethical standard of the Helsinki Declaration of 1975, and the revised version in 1983. We also applied the procedures from Workman et al. [29]
Patient tumor samples
Tumor tissue
Kindlin-2 is upregulated but HOXB9 is downregulated by TGF-β1 in PDAC cells
The relationship between Kindlin-2 and TGF-β1 in human cancer cells has not been reported [15], [16]. In this study, we examined the role of TGF-β1 in the regulation of Kindlin-2 expression in PDAC cells. Expression of Kindlin-2 was determined by Western blot analysis in PANC-1 cells treated with 5 ng/ml of TGF-β1 for the indicated time (Fig. 1A upper and lower panels and Supplementary Table S1) or treated with TGF-β1 at the indicated concentrations for 24 h (Fig. 1B upper and lower panels and
Discussion
The pancreas is derived from endoderm during human embryonic development and it expresses low levels of Kindlin-2 [37]. However, in PDAC specimens Kindlin-2 expression is significantly elevated in tumors compared with the normal pancreatic tissues adjacent to carcinoma, suggesting that Kindlin-2 may play a role in PDAC. In this report, we found that Kindlin-2 regulates PDAC progression by promoting PDAC cell proliferation, migration and invasion. Elevated Kindlin-2 levels in PDAC patients
Conflict of interest
None.
Acknowledgements
This work was supported by grants from the Ministry of Science and Technology of China 2015CB553906, 2013CB910501 and 2013ZX09401004-006, the National Natural Science Foundation of China 81230051, 81472734, 31170711, 81321003 and 30830048, the 111 Project of the Ministry of Education, Beijing Natural Science Foundation 7120002 and Peking University grants BMU20120314 and BMU20130364, and a Leading Academic Discipline Project of Beijing Education Bureau to H.Z. This work was also supported by
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