Since lung cancer is the leading cause of cancer-related death worldwide, research is being conducted to discover anticancer agents as its treatment. Eleutherine bulbosa, a Dayak folklore medicine, exhibited anticancer effects against several cancer cells; however, its anticancer potency against lung cancer cells has not been explored yet. This study aims to determine the anticancer potency of E. bulbosa bulbs against lung cancer cells (A549) using 2D and 3D culture models, as well as determine its active compounds using gas chromatography-mass spectrometry (GC-MS) analysis. Three fractions of E. bulbosa bulbs, namely chloroform, n-hexane, and ethyl acetate, were tested for cytotoxicity using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) and CellTiter-Glo. The antiproliferative effects of the most cytotoxic fraction against the 2D culture model were determined by a clonogenic survival assay and propidium iodide/Hoechst 33342 double staining, whereas the effects against the 3D culture model were determined by microscopy, flow cytometry, and gene expression analysis. The chloroform fraction is the most cytotoxic against A549 cells than other fractions, and it inhibited colony formation and induced apoptosis of A549 cells. The chloroform fraction also inhibited the growth of the A549 spheroid by suppressing the spheroid size, inducing apoptosis, reducing the proportion of CD44 lung cancer stem cells, causing arrest at the S phase of the cell cycle, and suppressing the expression of the SOX2 and MYC genes. Furthermore, the GC-MS analysis detected 20 active compounds in the chloroform fraction, including the major compounds of eleutherine and isoeleutherine. In conclusion, the chloroform fraction of E. bulbosa bulbs exhibit its antiproliferative effect on 2D and 3D culture models of A549 cells, suggesting it could be a lung cancer chemopreventive agent.
Introduction: Pheromones are chemicals produced by an animal that affects the behavior of another animal or the same species. Information conveyed includes location, presence of food or threat, sexual attraction, courtship, and dam-pup interactions. Pheromones are used widely in laboratory mice facilities to synchronize estrus and simultaneous breeding for logistic purposes. Female mice housed together in the absence of the male exhibit the Lee-Boot effect of lengthened diestrus or ovarian inactive period of up to several weeks. Whitten effect is described when a large number of female mice housed together in the absence of the male and having diestrus, will enter estrous 48 to 72 hours later upon exposure to male odors or male mouse urine soaked-bedding. Objective: The aim of this study is to determine the time taken for the Whitten effect to occur based on changes in vaginal cell characteristics, vulva appearance and behavior in grouped female mice. Methodology: Ten female mice were acclimatized to the animal facility for 3 estrus cycles or 12 days. Phases of the estrus cycle were evaluated by visual observation to assess changes to the vulva and vaginal cytology. Male urine soaked-bedding were exposed to females for 3 days and the time taken for the Whitten effect to occur was determined based on changes in vaginal cell characteristics, vulva appearance and observation of behavior. Result: The Cochran's Q test was used to observe the changes from diestrus to proestrus and later estrus. The results showed a significant difference (p<0.05) in the number of mice that successfully enter the proestrus and estrus phases over a four time point, χ 2 (2) = 18.857. Conclusion: The Whitten effect occurs after 72 hours of exposure to male urine soaked-bedding based on vulva appearance, vaginal cell characteristics and behavior in grouped female mice.
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