lithium-chloride and Uterine-Cervical-Neoplasms

The bioengineered luciferase reporter has been widely used for monitoring of a variety of molecular events in living cells because of their ability to provide highly sensitive quantitation with broad linearity. In the present study, we made a cyclin A2-luciferase (CYCA-Luc) fusion protein and examined the utility of this optical reporter for monitoring G2-phase cell cycle arrest in living animals. In vitro luciferase assay and in vivo bioluminescence imaging assay showed that the lithium chloride (LiCl), G2-phase-specific drug, induced G2-phase arrest of cell cycle and increased the activity of this reporter under in vitro or in vivo conditions, and this reporter can also be potentially used in high-throughput screening efforts aimed at discovering novel anti-cancer drugs that will cause cell cycle arrest at the G2-phase in cultivated cell lines and animal models.

Topics: Animals; Cyclin A2; Female; G2 Phase Cell Cycle Checkpoints; Genes, Reporter; HeLa Cells; High-Throughput Screening Assays; Humans; Lithium Chloride; Luciferases; Luminescent Measurements; Mice, Inbred BALB C; Mice, Nude; Optical Imaging; Recombinant Fusion Proteins; Uterine Cervical Neoplasms; Xenograft Model Antitumor Assays

Cervical cancer continues to threaten women's health worldwide, and the incidence of cervical adenocarcinoma (AD) is rising in the developed countries. Previously, we showed that glucose-regulated protein 58 (Grp58) served as an independent factor predictive of poor prognosis of patients with cervical AD. However, the molecular mechanism underlying the involvement of Grp58 in cervical carcinogenesis is currently unknown.. DNA microarray and enrichment analysis were used to identify the pathways disrupted by knockdown of Grp58 expression.. Among the pathway identified, the WNT signaling pathway was one of those that were significantly associated with knockdown of Grp58 expression in HeLa cells. Our experiments showed that β-catenin, a critical effector of WNT signaling, was stabilized thereby accumulated in stable Grp58 knockdown cells. Membrane localization of β-catenin was observed in Grp58 knockdown, but not control cells. Using a transwell assay, we found that accumulated β-catenin induced by Grp58 knockdown or lithium chloride treatment inhibited the migration ability of HeLa cells. Furthermore, an inverse expression pattern of Grp58 and β-catenin was observed in cervical tissues.. Our results demonstrate that β-catenin stability is negatively regulated by Grp58 in HeLa cells. Overexpression of Grp58 may be responsible for the loss of or decrease in membranous β-catenin expression in cervical AD.

Topics: Adenocarcinoma; beta Catenin; Female; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; HeLa Cells; Humans; Lithium Chloride; Oligonucleotide Array Sequence Analysis; Protein Disulfide-Isomerases; Protein Stability; Uterine Cervical Neoplasms; Wnt Signaling Pathway