SUMMARY
Cancer is one of the most serious health challenges the world in our modern era. Over the past two decades, this disease has been the second cause of death. Gastric cancer (GC) still comprises one of the major contributors to cancer prevalence and death around the world. Despite the great scientific progress in treatment methods, the success rate was limited, in addition to the side effects, as well as the high cost of these treatments. These reasons prompted researchers to seek for alternative treatments, such as using natural plant materials. Crataegus azarolus commonly called hawthorn that contains compounds with health potentials and minimal side effects. Extracts of hawthorn have been found to exert negative effects on the viability of many types of cancer through a number of pathways both in-vivo and in-vitro. The aim of this study was to identify and study a transcription factor expression Early Growth Response-1 (EGR1) pattern in GC through plant extracts, and to investigate the possible anti-proliferative, migration, and apoptotic effects of methanol and acetone extracts of C. azarolus fruits and leaves on human adenocarcinoma gastric (AGS) and human fibroblast cell lines. AGS cells were treated with different concentrations of both methanol and acetone fruits and leaves extracts (10, 50, 100, 250, and 500µg/ml). Our results demonstrated that the methanol leaves (ML) extract showed significant higher anti-proliferative at both 250 and 500µg/ml concentrations respectively, when estimated by MTT assay at p< 0.001 with an IC50 values 250µg/ml. In scratching assay, the results revealed that after 48h of treatments methanol fruits (MF) extract showed significant (p˂0.01) inhibitory effects on AGS cells migration comparing to the control, and acetone leaves (AL) extract exhibited an inhibitory effects on migration of AGS cells but it was not significant. Whereas, significant (p˂0.001) inhibitory effects on normal fibroblast cells showed after 48h and treatment with MF and AL extracts. Clonogenicity of AGS cells was significantly (p˂0.001) impaired and resulted in reduction in holoclone formation in acetone fruits (AF) and AL extracts. Apoptotic effects percentage increased when AGS cells were treated with MF, AF, and AL extracts of C. azarolus as compared with control. The levels of apoptosis were 16.8% untreated AGS cells, 31.4% MF, 16% ML, 37.9% AF, and 20.6% AL. RT-PCR showed significant (p˂0.001) upregulation in expression of EGR1 in AF extract, while EZH2 was significantly (p˂0.001) upregulated when treated with MF and AL. The tumor suppressor genes, PTEN and NDRG1, were significantly upregulated in MF and AF extracts respectively. In conclusion, C. azarolus had anti-cancer and cytotoxic effects on the AGS cell line and induced apoptosis in AGS cells. C. azarolus extracts might induce EGR1 and EZH2 independent apoptotic mechanism. Therefore, C. azarolus might be suggested as a therapeutic candidate for cancer treatment.
Key Words: Gastric cancer; Crataegus azarolus; human adenocarcinoma gastric (AGS); Extracts; Anti-proliferative; Apoptosis; EGR1.