GRIM-19 over-expression represses the proliferation and invasion of orthotopically implanted hepatocarcinoma tumors associated with downregulation of Stat3 signaling

Abstract

The retinoid-interferon-induced mortality-19 (GRIM-19) gene has been identified as a negative regulator associated with tumor development. The current study created a model of an orthotopically implanted hepatocarcinoma tumor to verify the inhibitory effect of GRIM-19 in vivo. After treatment with GRIM-19 carried by attenuated Salmonella, transplanted tumors were measured with an Imaging System. The expression of GRIM-19, Stat3/ p-Stat3, cyclinD1, CDK4, PCNA, Bax/Bcl-2, cleaved caspase-9/3, VEGF, and MMP-2/9 was determined using immunohistochemistry and Western blot analysis. The cell cycle was assessed using flow cytometry (FCM). Apoptosis was determined using FCM and a TUNEL assay. Results indicated that GRIM-19 overexpression resulted in inhibition of peritoneal metastasis, induction of cell cycle arrest, and apoptosis in vivo. In addition, the expression of Stat3/p-Stat3 was down-regulated by GRIM-19. These results suggest that GRIM-19 overexpression could suppress the growth of orthotopically implanted hepatocarcinoma tumors by reversing the regulation of the Stat3 signaling pathway. This approach could potentially be a powerful treatment for hepatocarcinoma.

Attenuated Salmonella carrying plasmid co-expressing HPV16 L1 and siRNA-E6 for cervical cancer therapy

“…However, since high-risk HPV E6/E7 proteins may lead to an aggressive proliferation in normal cells, inhibiting the expression of E6/E7 through RNA interference has been confirmed to achieve considerable treatment effects on cervical cancer 23 . Unfortunately, the absence of vector that stably transfers the genes into targeted cells limits the application of this technology 17 .…”

“…However, with continuous HPV16 infection, E6 exerts its ubiquitin-mediated p53-degration effects by interacting with E6AP in cervical cells 33 . Bax protein and Caspase family proteins are crucial mediators and symbols of cell apoptosis regulated by P53 17 , 34 . c-Caspase9 and c-Caspase3 can trigger the caspase cascade and process of apoptosis 35 .…”

“…Western blot was performed as described in previous study 17 . Rabbit anti-HPV16-L1, anti-HPV16-E6, anti-HPV16-E7 (R&D, USA), Rabbit anti-P53, anti-Bax, anti-Caspase9, anti-Caspase3, anti-cleaved-Caspase3 and anti-β-actin (Affinity, China) were used as the primary antibodies.…”

“…However, the absence of vectors that can stably transmit siRNA into target cells limits their clinical applications 16 . Attenuated Salmonella can easily accumulate and proliferate in the tumor microenvironment 17 . Moreover, alive attenuated Salmonella can induce mucosal immune response 10 .…”

Analysis of the association between KIN17 expression and the clinical features/prognosis of epithelial ovarian cancer, and the effects of KIN17 in SKOV3 cells

“…Immunohistochemistry. Immunohistochemical (IHC) staining was performed as described previously (25). Briefly, after being fixed in 10% formalin at room temperature for 24 h, EOC tissues were embedded in paraffin and cut into 3-µm thick sections.…”