Silencing of Girdin suppresses the malignant behavior of colorectal carcinoma cells

Abstract

The aim of the present study was to investigate the effect of the actin‑binding protein Girdin on the proliferation, invasion and migration of colorectal cancer (CRC) cells. Cultured CRC cells (LoVo cell line) were transfected by Girdin‑specific and control shRNA constructs and analyzed for proliferation, invasion and migration by the MTT, Transwell and wound‑healing assays, respectively. The activation of the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway and expression of proinflammatory cytokines was examined by western blotting and ELISA assay, respectively. The effect of Girdin silencing on CRC growth was also evaluated in a xenograft model using nude mice, which were subcutaneously injected with Girdin‑deficient and negative control LoVo cells and analyzed for tumor volume and weight. Transfection of LoVo cells with Girdin‑specific shRNA inhibited Girdin mRNA expression to 27.5% and protein expression to 36.7% when compared with expression levels in the control cells (P<0.001) and significantly demonstrated suppression of LoVo cell proliferation (P<0.05), invasion (P<0.01) and migration (P<0.01). Furthermore, Girdin silencing downregulated the phosphorylation of the signaling proteins JAK (by 42%, P<0.001) and STAT3 (by 34%, P<0.01) and the content of IFN (by 28%, P<0.001) and IL‑6 (by 44%, P<0.001) compared to the control. Notably, inhibition of Girdin expression effectively suppressed tumorigenicity of LoVo cells in vivo as evidenced by the reduced volume (P<0.05) and weight (P<0.05) of the tumors derived from Girdin shRNA‑transfected LoVo cells compared to those from the control cells. In conclusion, the silencing of Girdin expression inhibited the malignant behavior of CRC cells via the downregulation of the JAK/STAT signaling pathway, indicating Girdin as a potential therapeutic target in CRC. In the present study, we revealed, for the first time, that the malignant behavior of CRC cells depended on the expression of an actin‑binding protein, Girdin. Silencing of Girdin expression by specific shRNA suppressed the proliferation, invasion, and migration of CRC cells through the decrease in proinflammatory cytokines IFN and IL‑6 and the downregulation of the JAK/STAT signaling pathway. Our findings indicated that Girdin expression may be a potential novel therapeutic target in CRC.

The stress polarity signaling (SPS) pathway serves as a marker and a target in the leaky gut barrier: implications in aging and cancer

“…It is important to note that the pattern of pS245GIV staining is strikingly different from the levels of total GIV during CRC initiation and progression; we have previously shown that GIV mRNA and protein generally decreases first during polyp formation and progression and subsequently increases during CRC progression to metastasis . We and others have shown that increased levels of GIV in established CRCs correlate with aggressive features (Garcia-Marcos et al, 2011b;Jun et al, 2013;Aznar et al, 2016;Barbazan et al, 2016;Ghosh et al, 2016;Lu et al, 2018), for example, shorter metastasis-free survival, chemoresistance, and stemness, largely attributed to its ability to scaffold multi-receptor signaling cascades and enhance them via G protein intermediates to trigger epithelial mesenchymal transition (reviewed in Ghosh (2015)). Taken together, the loss of pS245 GIV early during polyp to CRC progression underscores how GIV may serve as a tumor suppressor when localized to TJs in the polarized normal epithelium before it assumes its role as an enhancer of epithelial-mesenchymal transition and stemness in established CRCs.…”

Girdin regulates both migration and angiogenesis in pancreatic cancer cell lines

“…Girdin is highly expressed in various human malignancies, including breast, colon, lung, thyroid and cervical cancer (37)(38)(39)(40). The Department of Gastroenterological Surgery, Nagoya City University Graduate School of Medical Sciences also reported that high Girdin expression was present in esophageal cancer and it is correlated with invasive potential and prognosis (14).…”

Roles of metformin‐mediated girdin expression in metastasis of epithelial ovarian cancer

“…Girdin is highly expressed in serval cancers (Lu, et al, 2018, Shibata et al, 2013, Wang et al, 2017), and downregulation of Girdin suppressed colorectal cancer cell migration and invasion (Lu et al, 2018). Consistence with the reported studies, herein, we found that Girdin was upregulated in human EOC (Figure 1), and knockdown of Girdin inhibited migration and invasion of EOC cells (SKOV3 and A2780), and retarded the expressions of MMP2/9 (invasive and metastatic factors), however, overexpression of Girdin resulted in the opposite (Figures 2 and 3).…”

“…Evidence suggests that Girdin is a regulator of cell motility and implicated in cancer cell migration, particularly through enhancing the activity of AKT (or phosphoinositide 3‐kinase [PI3K]‐Akt signal transduction pathway), which plays a core role in EOC cell migration and polarization (Mullerova et al, Wang et al, 2017, Weng et al, 2010). Girdin is highly expressed and emerged as therapeutic target in colorectal cancer, breast cancer as well as esophageal squamous cell carcinoma (Lu et al, 2018, Shibata et al, 2013, Wang, et al, 2017). However, whether and how does Girdin function in human EOC remains largely unknown.…”