2. Type 2 diabetes
mellitus
Is the most common endocrine disease. Is
a serious, complex chronic condition
characterized by hyperglycaemia and
insulin resistance.
3. ecc DNAs
Increased SORBS1 eccDNA circle is
associated with insulin resistance.
SORBS1, a novel eccDNA, is
validated as the most significant
up-regulated eccDNA in serum
from patients with DM2.
Are a group of novel circular DNAs,
physically derived from genomic DNA.
They contribute to oncogene plasticity
and drug resistance.
SORBS1circle
4. Evaluate the increased serum
extrachromosomal circular DNA
SORBS1circle level associated
with insulin resistance in
patients with newly diagnosed
type 2 diabetes mellitus
General objective
5. METHODS
CIRCLE SEQUENCING QUANTITATIVE PCR
Peripheral venous blood (5 ml) was collected
using vacutainer serum tubes from fasting
participants of at least 12 h, DNA extractions
were performed For linear DNA removal and
eccDNA enrichment, DNA was sequenced to
align with the reference genome to detect
eccDNA in all samples to obtain read counts with
EdgeR software (v0.6.9) to normalize and identify
differentially expressed ecDNA filtered by p-
value..
it was done by quantitatively and specifically
amplifying the proteins that encode serum
extrachromosomal circular DNA SORBS1
6. METHODS
WESTERN BLOT FISH
The obtained proteins were separated by SDS-
PAGE and immediately transferred to
nitrocellulose membranes. The membranes
were incubated with primary antibodies
overnight and then with appropriate secondary
antibodies for 2 hours. The antibodies used
were phosphorylated protein kinase B, total Akt,
Bcl-2 and Bax.
HepG2 cells were first fixed with 4%
paraformaldehyde for 10 min at room temperature
and treated with 0.5% Triton X-100 for 20 min at 4
℃ . After washing with PBS buffer three times, the
cells were treated with prehybridization buffer at 37
℃ for 30 min, and then incubated with hybridization
buffer at 37 ℃ overnight. After incubation, the cells
were washed with 4 × saline, sodium citrate with
0.1% Tween-20 three times at 42 ℃, followedcon 2
× SSC, 1 × SSC at 42 ℃ and then with PBS once at
room temperature. The nuclei were labeled with DAPI
9. CONCLUSION
AUTHOR
The host gene SORBS1 is an
important adaptor protein in the
signaling pathway of insulin-
stimulated glucose uptake
CA Bauman
Qin Yang
Siqian Gong
OPINION
Insulin resistance occurs when
target tissues, such as the liver,
become less sensitive
to the effects of insulin
Genetic variation of SORBS1 gene
is associated with glucose
homeostasis of diabetes
10. CONCLUSIONS
The identification of changes in eccDNA expression in
T2DM patients, as well as the characterization of
eccDNA SORBS1circle as a specific marker, highlights
the importance of understanding the underlying
molecular mechanisms in the pathogenesis of diabetes
and provides a basis for the development of new
diagnostic and therapeutic strategies
The ability to identify and validate specific eccDNAs, such
as SORBS1circle, as potential biomarkers highlights the
crucial role of molecular biology in identifying new
therapeutic targets and developing more precise and
personalized diagnostic tools for metabolic diseases like
diabetes
