Previously, caspase-1 was implicated in the execution of apoptosis. a large amount of genomic and epigenetic alterations has been recognized, mainly centered on apoptosis, autophagy and hepatitis virus, the treatment of HCC is still a conundrum [24]. Therefore , it is very important to clarify the molecular mechanisms of HCC and find novel therapeutic focuses on for this malignancy. Pyroptosis is a form of caspase-1 dependent programmed cell death described in myeloid cells. Activation of caspase-1 distinguishes pyroptosis from other kinds of cell death. In this process, cells recognize certain harmful signals, produce cytokines, swell, burst, and ultimately die [5]. Pyroptosis has been demonstrated to worsen diabetic cardiomyopathy, hepatic fibrosis and diabetes [68]. For cancer cells, the activation of CK-666 pyroptosis may promote cell death and thus exert anticancer properties. The loss of caspase-1 gene expression was found in human being prostate cancer [9, 10]. Moreover, caspase-1 deficiency enhances inammation- induced colorectal tumor formation in mice [11]. However , little is known regarding the role of pyroptosis in HCC. Berberine is a natural isoquinoline alkaloid, which is well known for its excellent antimicrobial activity and mild side effects [12]. Recently, an increasing number of studies has proved that berberine is a promising agent intended for cancer treatment [13]. It has been reported that berberine can induce both autophagy and apoptosis in HCC cells [14, 15]. In our preliminary experiments, we observed that berberine could activate caspase-1. We therefore set out the present study and explored the role of pyroptosis in the progression of HCC and the involvement of pyroptosis as a possible mechanism by which berberine induces HCC. == RESULTS == == Lack of caspase-1 expression in HCC == The expression Rabbit Polyclonal to AKAP4 of caspase-1 in HCC was decided in human being tissues and cell lines. A significant decrease in caspase-1 immunostaining was observed in CK-666 HCC tissues from patients (Figure1A). To further evaluate the expression of caspase-1, real-time PCR and western blot were performed. The results CK-666 showed that caspase-1 mRNA and protein levels were downregulated in HCC tissues relative to adjacent normal tissues (Figure1B&1C). A similar phenomenon was observed in HCC and hepatocyte cell lines: the expression of caspase-1 in Bel-7402 and HepG2 cells was lower than that in HL-7702 (Figure1D, E&1F). These findings suggested that loss of caspase-1 expression is involved in the pathogenesis of HCC. == Determine 1 . Lack of caspase-1 expression in human being hepatocellular carcinoma (HCC) tissues and cells. == A. The immunohistochemical staining of caspase-1 in HCC and adjacent normal tissues. B&C. The expression of caspase-1 CK-666 mRNA and protein in CK-666 HCC and surrounding normal tissues. D. The immunofluorescence staining of caspase-1 in HL-7702, Bel-7402 and HepG2 cells. E&F. The expression of caspase-1 mRNA and protein in HL-7702, Bel-7402 and HepG2 cells. GAPDH served because an internal control. For panel B and C, *P < 0. 05 versus Normal, **P < 0. 01 versus Normal. Intended for panel E & F, *P < 0. 05 versus HL-7702, **P < 0. 01 versus HL-7702, ***P < 0. 001 versus HL-7702; n= a few. == Berberine induces pyroptosis in HepG2 cells == The cell number was decreased and cell swelling was observed after treatment with berberine (Figure2A). Berberine increased the mRNA and protein expression of caspase-1 in HepG2 cells in a concentration-dependent manner with statistical significance starting from 50 M (Figure2B&2C). Thus, this concentration was used for the subsequent experiments. We also found that berberine decreased the viability.