Curcumin induces re­expression of BRCA1 and suppression of γ synuclein by modulating DNA promoter methylation in breast cancer cell lines.

Restoration of normal DNA promoter methylation and expression states of cancer­related genes may be an option for the prevention as well as the treatment of several types of cancer. Constitutional promoter methylation of BRCA1 DNA repair associated (BRCA1) gene is linked with a high risk of developing breast and ovarian cancer. Furthermore, hypomethylation of the proto­oncogene Î³ synuclein (SNCG) is associated with the metastasis of breast and ovarian cancer and reduced disease­free survival (DFS). In the present study, we evaluated the potential of curcumin to re­express hypermethylated BRCA1 and to suppress hypomethylated SNCG in triple­negative breast cancer (TNBC) cell line HCC­38, the estrogen receptor­negative/progesterone receptor­negative (ER­/PR­) cell line UACC­3199, and the ER+/PR+ cell line T47D. The cells were treated with 5 and 10 µM curcumin for 6 days and with 5­aza­2'­deoxycytidine (5'­aza­CdR) for 48 h. Methylation­specific PCR and bisulfite pyrosequencing assays were used to assess DNA promoter methylation while gene expression levels were analyzed using quantitative real­time PCR and immunoblotting. We found that curcumin treatment restored BRCA1 gene expression by reducing the DNA promoter methylation level in HCC­38 and UACC­3199 cells and that it suppressed the expression of SNCG by inducing DNA promoter methylation in T47D cells. Notably, 5'­aza­CdR restored BRCA1 gene expression only in UACC­3199, and not in HCC­38 cells. Curcumin­induced hypomethylation of the BRCA1 promoter appears to be realized through the upregulation of the ten­eleven translocation 1 (TET1) gene, whereas curcumin­induced hypermethylation of SNCG may be realized through the upregulation of the DNA methyltransferase 3 (DNMT3) and the downregulation of TET1. Notably, miR­29b was found to be reversely expressed compared to TET1 in curcumin­ and 5'­aza­CdR­treated cells, suggesting its involvement in the regulation of TET1. Overall, our results indicate that curcumin has an intrinsic dual function on DNA promoter methylation. We believe that curcumin may be considered a promising therapeutic option for treating TNBC patients in addition to preventing breast and ovarian cancer, particularly in cancer­free females harboring methylated BRCA1.

Incidence of hypoglycemia and its risk factors among diabetics during Ramadan in Abha city, Aseer Region, KSA.

BACKGROUND AND AIMS: This study aims to explore the incidence of hypoglycemia and its risk factors among diabetic patients attending primary health care center during Ramadan Abha city, Aseer region, Saudi Arabia. METHODS: This cross-sectional study was conducted among adult diabetic patients attending Primary health care centers (PHCCs) in Abha city, southwest of KSA. A questionnaire in Arabic language was used. It consisted of five parts that covered patients demographic and DM relevant profile, hypoglycemia attacks during Ramadan, compliance with drug, diet, exercise and glucose monitoring. Four PHCCs in Abha city were selected randomly to conduct this study. All diabetic patients who attended the selected PHCCs during the month of Shawwal 1439 (corresponding to June-July 2018) were interviewed by the investigators. Data were coded, entered and analyzed using SPSS version 22. Appropriate statistical tests were used accordingly and P value was considered as significant if it was less than 5%. RESULTS: The total patients participated in this study was 378. The mean age was 45 years, males represents 51%, mean duration of DM was 12 years, type-1DM constitutes about one third. Most of type-1 DM patients used act rapid and long acting insulin (65%), while in type-2 DM, more than one third (38%) used OHA, 8% were on insulin alone. More than half of patients (52%) reported at least one attack of hypoglycemia during Ramadan, (29%) out of them had more than four attacks. About two third of attacks (67%) occurred in the morning and evening while less than one fourth have hypoglycemia at night (17%), (2%) visited ER or PHC and 1% were admitted to hospital for further management. CONCLUSION: This study revealed that the incidence of hypoglycemia among diabetics was high. Many Risk factors were identified; young age, type-1 DM, long duration of DM, insulin use. More attacks occurred during Ramadan day period and led to breaking the fasting among all affected patients. Most of patients were not given instructions regarding self-care immediately before or during Ramadan. Structured health education program for diabetics attending PHCC should be constructed and implemented before beginning of Ramadan in order to minimize the incidence of acute complications particularly hypoglycemia.

Methylation of BRCA1 and MGMT genes in white blood cells are transmitted from mothers to daughters.

BACKGROUND: Constitutive methylation of tumor suppressor genes are associated with increased cancer risk. However, to date, the question of epimutational transmission of these genes remains unresolved. Here, we studied the potential transmission of BRCA1 and MGMT promoter methylations in mother-newborn pairs. METHODS: A total of 1014 female subjects (cancer-free women, n = 268; delivering women, n = 295; newborn females, n = 302; breast cancer patients, n = 67; ovarian cancer patients, n = 82) were screened for methylation status in white blood cells (WBC) using methylation-specific PCR and bisulfite pyrosequencing assays. In addition, BRCA1 gene expression levels were analyzed by quantitative real-time PCR. RESULTS: We found similar methylation frequencies in newborn and adults for both BRCA1 (9.9 and 9.3%) and MGMT (12.3 and 13.1%). Of the 290 mother-newborn pairs analyzed for promoter methylation, 20 mothers were found to be positive for BRCA1 and 29 for MGMT. Four mother-newborn pairs were positive for methylated BRCA1 (20%) and nine pairs were positive for methylated MGMT (31%). Intriguingly, the delivering women had 26% lower BRCA1 and MGMT methylation frequencies than those of the cancer-free female subjects. BRCA1 was downregulated in both cancer-free woman carriers and breast cancer patients but not in newborn carriers. There was a statistically significant association between the MGMT promoter methylation and late-onset breast cancers. CONCLUSIONS: Our study demonstrates that BRCA1and MGMT epimutations are present from the early life of the carriers. We show the transmission of BRCA1 and MGMT epimutations from mother to daughter. Our data also point at the possible demethylation of BRCA1and MGMT during pregnancy.