Isolation and identification of Lactobacillus from Gilan traditional yogurt and the effect of its extract on p۲۱, p۵۳ and casp۳ signaling pathways in MCf-۷ breast cancer cell line

BACKGROUND AND OBJECTIVESToday, probiotics are used as supplements and have been increasingly noticed in recent years. Several studies have shown the ability of probiotics to prevent and treat various diseases, like cancer. Probiotics exert their anti-cancer effects through anti-inflammatory, antioxidant, anti-angiogenic activities. Therefore, they can be proposed as a rational approach for the prevention and treatment of cancer, including breast cancer. Therefore, the focus of this study is to investigate the effects of probiotics isolated from dairy products on MCF-۷ breast cancer cells .MATERIALS AND METHODSVarious yogurt samples using sterile saline and then cultured in MRS agar medium. Then lactobacilli are identified using microbial and biochemical tests. Then, with the help of ۱۶srRNA Lactobacillus specific primers, DNA extraction and PCR are carried out to determine the identity. Then, the identified Lactobacillus extract was determined by the MTT assay in order to determine the best effective dilution on the cells. Then, the extract with appropriate dilution was applied on HT-۲۹ cancer cells. RNA extraction was performed for three hours and six hours after the effect of the extract on cancer cells. Then cDNA synthesis was done using the kit according to the protocol. In order to perform Real time PCR, primer design was performed on p۲۱, p۵۳ and casp۳ genes .RESULTS AND DISCUSSIONLocal dairy samples were collected from different regions of Gilan province and lactobacilli were isolated using MRS-Agar medium. In order to molecularly confirm the isolated from local yogurt samples as probiotic strains, DNA extraction was done using a kit in order to determine the amount of apoptosis induced in MCf-۷ cancer cell line by Lactobacillus extract, respectively for gene p۲۱ equal to ۲.۷۷ and ۲.۴۲ for p۵۳ gene and ۲.۳۲ for casp۳ gene and ۱ for GAPDH control gene. According to the results, the effect of Lactobacillus extract on MCf-۷ cancer cells increases the expression of all three genes, which indicates an increase in apoptosis.CONCLUSIONThe results of this study showed that by adding lactobacillus, the expression of p۲۱, p۵۳ and casp۳ genes increases, which by activating the apoptosis pathway leads cancer genes to programmed death and can be used as a complementary treatment

کلیدواژه ها:

Lactobacillus ، traditional Gilan yogurt ، p۲۱ ، p۵۳ and casp۳ ، MCf-۷ ، breast cancer