Effect of betulinic acid on the regulation of Hiwi and cyclin B1 in human gastric adenocarcinoma AGS cells

Abstract

Aim:To investigate the effect of betulinic acid (BA) on the proliferation, apoptosis and cell cycle of gastric adenocarcinoma cell AGS in vitro and elucidate the underlying mechanisms. Methods: The effect of BA on the proliferation of AGS cells was measured by using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) assay. Apoptosis was analyzed by using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double-labeled flow cytometry (FCM) and Hoechst 33258 staining. The influence of BA on cell cycle of AGS cells was tested by PI staining. Both FCM and reverse transcription-PCR (RT-PCR) technologies were applied to detect the expression of Hiwi and Cyclin B1. Results: BA exhibited significant cell proliferation inhibition, as well as its potency of inducing apoptosis in AGS cells in vitro in a timeand dose-dependent manner. The IC 50 value for 24 h was 18.25 μg/mL (95% confidence interval: 15.16 to 27.31 μg/mL). Cells treated with BA showed increased cell population in G 2 /M phase, with decreased S phase population. The expression of Hiwi and Cyclin B1 was down-regulated in BA-treated AGS cells in a dose-dependent manner. Conclusion: BA exerted potent effect on growth inhibition, G 2 /M cell cycle arrest and induction of apoptosis in AGS cells in vitro, possibly associated with the down-regulation of Hiwi and its downstream target Cyclin B1 expression. The potent antitumor capacity of BA suggested that it could be a promising new experimental anticancer agent in human gastric adenocarcinoma treatment.

Jamun (<i>Syzygium cumini </i>(L.)): A Review of Its Food and Medicinal Uses

“…It also decreased GLI1, GLI2, PTCH1, and IGF2 expression as well as hedgehog-response in vitro. It also caused retarded the growth of RMS-13 xenografts by causing apoptosis and down-regulating GLI1 expression without affecting the microvascular density, cell proliferation, and myogenic differentiation unaffected [143]; 4) Induces apoptosis through the mitochondrial pathway and inducing cytochrome c…”

Betulinic acid inhibits autophagic flux and induces apoptosis in human multiple myeloma cells in vitro

“…Multiple mechanisms have been reported for the potent anticancer activity of BA [21] . In a previous study, we demonstrated that BA induced cell cycle arrest and apoptosis in human gastric adenocarcinoma AGS cells through the downregulation of Hiwi and Cyclin B1 [17] . In the present study, BA inhibited cell proliferation in multiple myeloma KM3 cells and induced apoptosis in a dose-dependent manner.…”

“…Various mechanisms have been reported for the extensive anticancer activity of BA, including directly triggering mitochondrial membrane permeabilization [20] , regulating Bcl-2 family proteins [21] , downregulating Hiwi expression [17] and suppressing the activation of the STAT3 pathway through the induction of the protein tyrosine phosphatase SHP-1 [19] . However, there are no reports as to whether BA has any effects on cancer cell autophagy.…”

“…Betulinic acid (BA), a natural pentacyclic triterpene isolated from birch, exhibits a variety of biological activities, including anti-HIV, anti-HBV, anti-malarial [8][9][10] and potent anti-tumor properties in melanoma [11] , leukemia [12] , colon [13] , lung [14] , gliomas [15] , prostate carcinoma [16] , gastric adenocarcinoma [17] and multiple myeloma [18,19] . Various mechanisms have been reported for the extensive anticancer activity of BA, including directly triggering mitochondrial membrane permeabilization [20] , regulating Bcl-2 family proteins [21] , downregulating Hiwi expression [17] and suppressing the activation of the STAT3 pathway through the induction of the protein tyrosine phosphatase SHP-1 [19] .…”

Steroid receptor coactivator-3 is a pivotal target of gambogic acid in B-cell Non-Hodgkin lymphoma and an inducer of histone H3 deacetylation

“…DNA content of cells was analyzed using PI staining, followed by flow cytometry analysis (FACS Calibur, BD Biosciences, San Diego, CA, USA), as previously described (Yang et al, 2010).…”