Molecular characterization of senescence marker protein-30 gene promoter: Identification of repressor elements and functional nuclear factor binding sites

<p>Abstract</p> <p>Background</p> <p>Senescence marker protein-30 (SMP30), whose expression declines during aging in rat liver, has been proposed as an important aging marker. Besides apoptosis, SMP30 also protects cells against various other injuries by enhancement of membrane calcium-pump activity. The mechanism of this differential gene expression mechanism is not known. DNA-protein interactions, mutation analysis and luciferase reporter assay studies have been performed to elucidate the mechanism of transcriptional regulation of SMP30 gene.</p> <p>Results</p> <p>We have characterized up to -2750 bp of the promoter by DNA-protein interactions studies. Twenty eight transcription factor binding sites have been identified by DNase I footprinting and electrophoretic mobility shift assay (EMSA). Transient transfection of 5' and 3' -deleted promoter-reporter constructs and luciferase assay illustrated the region between -128/+157 bp is sufficient to drive promoter activity. We have mapped an essential regulatory region between -513 to -352 bp which causes a drastic decline of reporter activity. This region contains CdxA, GATA2 and SRY transcription factor binding sites. Individual mutation of these three sites showed increase in reporter activity. Mutation in SRY site (-403/-368) showed maximum increase in reporter activity among these three sites. Therefore, we suggest that SRY like protein may be acting as a strong repressor of SMP30 gene along with CdxA and GATA-2. We also report that mutation of both Sp1 (172/-148 bp) and a C/EBPβ (-190/-177 bp) transcription binding site located adjacent to each other on SMP30 gene promoter, causes a significant enhancement in reporter activity than individual mutation, thus may be causing the repression of SMP30 promoter activity.</p> <p>Conclusion</p> <p>These studies provide novel insights into the mechanism that regulate SMP30 gene expression.</p

Quality Of Life and Coping Strategies Among Caregivers of Patient with Cardiovascular Diseases

Caregivers and family members of patients with long-term cardiovascular diseases often experience varying degrees of depression due to the significant changes in life style and the challenges associated with managing the condition. Adapting and modifying coping strategies as needed is crucial for caregivers to effectively decrease or manage stressful situations. The objectives of this study was to assess the quality of life and coping strategies among caregiver of cardiovascular patient. At IMS &amp; SUM Hospital in Bhubaneswar ,Odisha, a descriptive correlational research design was used. Purposive sampling selected 200 participants who completed a socio-demographic questionnaire. Two standardized scale s were employed: the WHOQOL-BREF scale to measure quality of life and the coping inventory for stressful situations to assess coping strategies. The study findings showed a significant negative correlation ( r = -0.338, p &lt; 0.01) between quality of life and coping strategies, as indicated by Karl Pearson’s correlation coefficient. This suggests that as, if the coping strategies is better, the quality of life is also better. Additionally, the study employed ANOVA and t-tests to examine the differences between quality of life and coping strategies. The study findings suggest that the quality of life of young individuals is more impacted by caregiving compared to the elderly. There is a significant negative correlation between quality of life and coping strategies, indicating that higher quality of life scores are associated with lower coping strategy scores. Conversely, better coping strategies are linked to better quality of life. Caregivers employ various coping strategies, and counselling can play a crucial role in providing emotional support, guidance, and practical advice to help caregivers manage challenges and maintain their well-being while caring for cardiovascular patients

3, 3′5 Triiodo L Thyronine Induces Apoptosis in Human Breast Cancer MCF-7cells, Repressing SMP30 Expression through Negative Thyroid Response Elements

Thyroid hormones regulate cell proliferation, differentiation as well as apoptosis. However molecular mechanism underlying apoptosis as a result of thyroid hormone signaling is poorly understood. The antiapoptotic role of Senescence Marker Protein-30 (SMP30) has been characterized in response to varieties of stimuli as well as in knock out model. Our earlier data suggest that thyroid hormone 3, 3'5 Triiodo L Thyronine (T(3)), represses SMP30 in rat liver.In highly metastatic MCF-7, human breast cancer cell line T3 treatment repressed SMP30 expression leading to enhanced apoptosis. Analysis by flow cytometry and other techniques revealed that overexpression and silencing of SMP30 in MCF-7 resulted in decelerated and accelerated apoptosis respectively. In order to identify the cis-acting elements involved in this regulation, we have analyzed hormone responsiveness of transiently transfected hSMP30 promoter deletion reporter vectors in MCF-7 cells. As opposed to the expected epigenetic outcome, thyroid hormone down regulated hSMP30 promoter activity despite enhanced recruitment of acetylated H3 on thyroid response elements (TREs). From the stand point of established epigenetic concept we have categorised these two TREs as negative response elements. Our attempt of siRNA mediated silencing of TRβ, reduced the fold of repression of SMP30 gene expression. In presence of thyroid hormone, Trichostatin- A (TSA), which is a Histone deacetylase (HDAC) inhibitor further inhibited SMP30 promoter activity. The above findings are in support of categorisation of both the thyroid response element as negative response elements as usually TSA should have reversed the repressions.This is the first report of novel mechanistic insights into the remarkable downregulation of SMP30 gene expression by thyroid hormone which in turn induces apoptosis in MCF-7 human breast cancer cells. We believe that our study represents a good ground for future effort to develop new therapeutic approaches to challenge the progression of breast cancer