We aimed to study the effects of LY294002, an inhibitor of class I phosphatidylinositol 3-kinase (PI3K), on proliferation, apoptosis, and autophagy in gastric cancer cell line SGC7901. In this study, we showed that LY294002 inhibited the viability of gastric cancer SGC7901 cells. We also showed that LY294002 increased the expression of microtubule-associated protein 1 light chain 3 (LC3), and increased monodansylcadaverine (MDC)-labeled vesicles. LY294002 activated autophagy by activating p53 and caspase-3, and induced apoptosis by up-regulating p53 and p53-up-regulated modulator of apoptosis (PUMA). Therefore, LY294002 might induce cytotoxicity in SGC7901 cells through activation of p53 and the downstream point PUMA. These findings suggest that inhibition of the class I PI3K signaling pathway is a potential strategy for managing gastric cancers.
Serotonin-synthesizing and serotonin-accumulating neurons were studied in the retinas of Xenopus laevis and Bufo marinus. All previously identified cell types exhibiting serotonin-like immunoreactivity (SLI) were labeled by intravitreal injection of 5,7-dihydroxytryptamine (5,7-DHT). They included two amacrine cell types (large and small) in both species, and one bipolar cell type in Xenopus. Incubation of retinas in culture medium in the ambient light reduced SLI in amacrine cells and enhanced the labeling in bipolar cells. After incubation, some photoreceptor cell bodies and large numbers of outer segments also displayed SLI in both species. Incubation with the serotonin-uptake inhibitor, fluoxetine, reduced immunolabeling in bipolar cells and outer segments to the level in the untreated retinas. Both large SLI and 5,7-DHT-accumulating amacrine cells in Xenopus and Bufo were labeled with an antibody raised against phenylalanine hydroxylase (PH), which binds to tryptophan 5-hydroxylase, one of the synthesizing enzymes for serotonin. Small SLI and 5,7-DHT-accumulating amacrine cells in both species represented two populations, one with and the other without PH-like immunoreactivity (PH-LI). The anti-PH antibody failed to label any SLI or 5,7-DHT-accumulating bipolar cells in Xenopus. These observations indicate that all large and some small SLI amacrine cells in the retinas of Xenopus and Bufo synthesize serotonin, while other small SLI amacrine, bipolar and photoreceptor cell bodies, and outer segments only accumulate serotonin.
The generation and changing distribution of neurons of the inner nuclear layer (INL) in the retina of two anuran species, Bufo marinus and Xenopus laevis, were studied from metamorphosis to adult. Morphometric studies were undertaken at six developmental stages in Bufo and four in Xenopus. The number and thickness of neurons in the INL were established in 29 predetermined retinal locations from serial sections of the eyes cut vertically or horizontally. The total number of neurons in the INL increased from metamorphosis to adult from 826,000 +/- 185 to 18,760,000 +/- 562 (mean +/- SD) in Bufo and from 308,000 +/- 25 to 877,000 +/- 31 in Xenopus. Over the same period the surface area of the INL increased about 50-fold from 2 mm2 to 96 mm2 in Bufo and 5-fold from 2.5 mm2 to 13 mm2 in Xenopus. In Bufo the difference between the highest cell number (central-temporal retina) and the lowest cell number in a sample area (dorsal and ventral peripheral retina) was 2.1:1 at metamorphosis. This ratio increased to 3.4:1 in the adult. Both the cell number and cell density per sample area in the INL was found to be higher along the nasotemporal meridian of the eye overlying the visual streak of the ganglion cell layer (GCL) of the retina. The retinal distribution of neurons in the INL did not change significantly during postmetamorphic growth in Xenopus. At metamorphosis a 1.7:1 difference was found between the highest neuron number (retinal ciliary margin) and lowest neuron number (retinal centre) decreasing to 1.5:1 in the adult. Retinae were labelled with 3H-thymidine in 15 mm Bufos and examined 2, 6, 12 and 18 weeks later. Higher rates of cell addition to the nasal and temporal poles of the INL were found compared with that at the dorsal and ventral poles. The retinal radial growth at the ciliary margin of the dorsal, ventral, nasal and temporal poles between the time of isotope injection and 18 weeks survival was found to be uneven; more radial elongation occurred at the nasal, dorsal and ventral poles and less at the temporal pole. These observations suggest that (a) the neuron distribution of the INL in adult animals approximates that of the GCL and (b) the visual streak-like area of the INL in Bufo develops by a sustained differential cell addition at the temporal and nasal poles of the retina.
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