cytochrome-c-t and Gallbladder-Neoplasms

To evaluate the effects of artemisinin on proliferation, cell cycle and apoptosis of gallbladder cancer cells.. Gallbladder carcinoma cell lines(GBC-SD and NOZ)were cultured in vitro. The effects of artemisinin in different concentration on proliferation of the two cell lines in vitro were examined using MTT assay. The cell cycle distribution of GBC-SD and NOZ cells 24 h after treatments with artemisinin(20 μmol/L) were examined using flow cytometry. The apoptosis of GBC-SD and NOZ cells 24 h after treatments with artemisinin (20 μmol/L) were examined using Annexin V/PI staining.The expressions of p-ERK1/2, CDK4, cyclin D1, p16, cytochrome C and caspase-3 were examined by Western blot assay. t-test and one way ANOVA were used to evaluate the differences between two groups and more than two groups, respectively.. The cell proliferation was significantly inhibited by artemisinin, the IC50 of artemisinin against GBC-SD and NOZ cells were 14.05 μmol/L and 12.42 μmol/L, respectively.Artemisinin induced cycle arrest, and G1 population of GBC-SD and NOZ cells increased to 74.60% and 78.86%. Cell apoptosis and apoptotic population of GBC-SD and NOZ cells were increased to 15.67% and 16.51% after dealt with artemisinin, respectively. In addition, expression of p16 was increased, and expressions of p-ERK1/2, CDK4 and cyclin D1 were down-regulated by artemisinin(all P<0.05). Cytochrome C was released from mitochondria to cytoplasm leading to the activation of caspase-3 and PARP after dealt with artemisinin(P<0.05).. The inhibition effect of artemisinin on the proliferation gallbladder cancer cells is accompanied by down-regulation of ERK1/2 signaling pathway, G1 phase arrest and triggering caspase-3-mediate apoptosis.

Topics: Apoptosis; Artemisinins; Caspase 3; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Cyclin-Dependent Kinase 4; Cytochromes c; Gallbladder Neoplasms; Humans; MAP Kinase Signaling System; Mitochondria

The biological function of tumor suppressor deleted in liver cancer 1 (DLC1) has been investigated in several types of human cancer, but its role in gallbladder cancer (GBC) is yet to be determined. In this research, we conducted in vitro and in vivo analysis to evaluate the inhibitory activities of DLC1 gene against GBC growth.. DLC1 expression in GBC tissues and cell lines was examined by immunohistochemical staining, reverse transcription polymerase chain reaction, and Western blot assay. The in vitro and in vivo effects of ectopic DLC1 expression on cell growth were evaluated. In addition, the effects of ectopic DLC1 expression on cell cycle, apoptosis, and migration were also evaluated. The expressions of cell cycle-related and apoptosis-related proteins were examined.. The downregulation of DLC1 expression was a common event in GBC tissues and cell lines. Restoration of DLC1 expression in GBC-SD and NOZ cells significantly reduced cell proliferation, migration in vitro, and the ability of these cells to form tumors in vivo. Restoration of DLC1 expression arrested GBC-SD and NOZ cells in G0/G1 phase through inducing p21 in a p53-independent manner. In addition, restoration of DLC1 expression induced extrinsic and intrinsic apoptotic pathway through promoting the expressions of Fas L/FADD, Bax, cytochrome c, cleaved caspase-8, -9, -3, and cleaved poly-(ADP-ribose) polymerase and suppressing bcl-2 expression in GBC-SD and NOZ cells.. Our findings suggested that dysregulated expression of DLC1 is involved in proliferation and invasion of GBC cells and may serve as a potential therapeutic target.

Topics: Apoptosis; bcl-2-Associated X Protein; Caspases; Cell Cycle; Cell Line, Tumor; Cell Movement; Cell Transformation, Neoplastic; Cyclin-Dependent Kinase Inhibitor p21; Cytochromes c; Fas Ligand Protein; Gallbladder Neoplasms; Gene Expression Regulation, Neoplastic; GTPase-Activating Proteins; Humans; In Vitro Techniques; Molecular Targeted Therapy; Tumor Suppressor Protein p53; Tumor Suppressor Proteins

Forkhead box L1 (FOXL1), considered as a novel candidate tumor suppressor, suppresses proliferation and invasion in certain cancers. However, the regulation and function of FOXL1 in gallbladder cancer (GBC) remains unclear.. FOXL1 expression at mRNA and protein levels in GBC tissues and cell lines were examined by RT-PCR, immunohistochemistry and western blot assay. FOXL1 expression in GBC cell lines was up-regulated by transfection with pcDNA-FOXL1. The effects of FOXL1 overexpression on cell proliferation, apoptosis, migration and invasion were evaluated in vitro or in vivo. In addition, the status of mediators involved in migration, invasion and apoptosis was examined using western blot after transfection with pcDNA-FOXL1.. FOXL1 was frequently downregulated in GBC tissues and cell lines. Its higher expression is associated with better prognosis, while its lower expression is correlated with advanced TNM stage and poor differentiation. FOXL1 overexpression in NOZ cells significantly suppresses cell proliferation, migration and invasion in vitro and tumorigenicity in nude mice. FOXL1 overexpression disrupted mitochondrial transmembrane potential and triggered mitochondria-mediated apoptosis in NOZ cells. In addition, FOXL1 overexpression suppressed ZEB1 expression and induced E-cadherin expression in NOZ cells.. Our findings suggested that dysregulated FOXL1 is involved in tumorigenesis and progression of GBC and may serve as a predictor of clinical outcome or even a therapeutic target for patients with GBC.

Topics: Aged; Animals; Apoptosis; Cadherins; Caspases; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cytochromes c; Down-Regulation; Female; Forkhead Transcription Factors; Gallbladder Neoplasms; Gene Expression Regulation, Neoplastic; Humans; Male; Membrane Potential, Mitochondrial; Mice, Nude; Middle Aged; Mitochondria; Neoplasm Invasiveness; RNA, Messenger

In this study, the anticancer effect of cirsimaritin, a natural flavonoid, against human gallbladder carcinoma cell line GBC-SD and the underlying mechanisms were investigated. Cirsimaritin inhibited the growth of tumor cells and induced mitochondrial apoptosis in GBC-SD cells. In addition, cirsimaritin triggered endoplasmic reticulum (ER) stress and down-regulated the phosphorylation of Akt, while knock-down of CHOP dramatically abrogated the inactivation of Akt and reversed the pro-apoptotic effect of cirsimaritin. Furthermore, cirsimaritin provoked the generation of reactive oxygen species in GBC-SD cells, while the antioxidant N-acetyl cysteine almost completely blocked the activation of ER stress and apoptosis, suggesting cirsimaritin-induced reactive oxygen species is an early event that triggers ER stress mitochondrial apoptotic pathways in GBC-SD cells.

Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Caspases; Cell Line, Tumor; Cytochromes c; Endoplasmic Reticulum; Flavones; Gallbladder Neoplasms; Humans; Membrane Potential, Mitochondrial; Mitochondria; Proto-Oncogene Proteins c-akt; Reactive Oxygen Species; Transcription Factor CHOP