Inhibition of Period Gene Expression Causes Repression of Cell Cycle Progression and Cell Growth in the Bombyx mori Cells

In this study, an animal model with a single BmPer gene product was selected to investigate the effect of Per-KD on the cell cycle and avoid the interaction of multiple Per expression products. There have been no previous reports of cell cycle changes after simultaneous knockdown or knockout of all mPer genes. A slow growing developmental model expressing a single BmPer gene that was continually knocked down in BmN cells (Per-KD) was used in this study. The BmN cells were free of endocrine influences. We compared cell proliferation and programmed cell death (PCD) and investigated the regulatory mechanisms in mutant and wild-type BmN cells. Materials and Methods Cell Preparation A wild-type (WT) B. mori ovary cell line (BmN) and a mutant line with stable interference of the BmPer gene (Per-KD) (Tao et al., 2017), were maintained in our laboratory and cultured in Grace insect medium (11605094, GIBCO, United States) with 10% (v/v) fetal bovine serum (FBS) (04-121-1A; Biological Industries, United States) at 26°C in the dark. The medium for culture of Per-KD cells included 0.05 mg/mL Zeocin (R25001, Invitrogen, United States). As shown in Figure 1, cell lines were synchronized by 24 h culture in serum-free Grace insect medium. The medium was then replaced with Grace insect medium with 10% FBS (v/v). The cells were counted and adjusted to the desired concentration. The time at which the synchronization process ended was recorded as time 0 h after synchronization. FIGURE ...
Source: Frontiers in Physiology - Category: Physiology Source Type: research

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