This study examined the relationship between microRNA-21 (miR-21), phosphatase and tensin homolog (PTEN) expression, and liver fibrosis in patients with chronic hepatitis C (CHC). The study found that PTEN expression was significantly lower and miR-21 expression was significantly higher in CHC patients compared to healthy controls. Lower PTEN expression and higher miR-21 expression were associated with more severe liver fibrosis. A predictive model was developed using PTEN expression, miR-21 expression, and total bilirubin that can identify CHC patients with higher fibrosis with high sensitivity and specificity.

1. MICRORNA-21 AND PTEN: A SIMPLE
PREDICTIVE MODEL FOR FIBROSIS IN CHC
PATIENTS.
Soumya J. Chowdhury*1, Vijay Karra1, Phani Gumma1, Premashis
Kar1
1Medicine, Maulana Azad Medical College, New Delhi,India, New
Delhi, India
OP044

2. There are no relevant and potential conflicts of interest.
Disclosure statement

3. CHRONIC HEPATITIS C AND FIBROSIS
 Hepatitis C virus (HCV) continues to be a major disease burden on the
world.
The worldwide estimated prevalence is about 3% .
The noted prevalence of hepatitis C in India is <2% .
Prospective studies have shown that 60 to 85 % of HCV-infected persons
develop chronic infection.
Fibrosis or scarring arises as a result of wound repair and leads to
cirrhosis, nodule formation, altered hepatic function and risk of liver-related
morbidity and mortality.
There has been evidences that fibrosis progression correlates with host
factors more than viral factors.

4. PTEN : ITS ROLE AND REGULATION
 PTEN- Phosphatase And Tensin-Like protein. Recommended name is
Phosphatidylinositol-3,4,5-triphosphate-3-phosphatase and dual
specificity protein phosphatse PTEN
 Regulates intracellular levels of Phosphatidylinositol-3,4,5-
triphosphate in cells and acts as a tumor suppressor by negatively
regulating AKT/PKB signaling.
 Expression or function of PTEN has found to be regulated by factors
like core proteins of HCV, micro RNA and have found to be involved in
many carcinoma.
Micro RNA are small RNAs that regulate gene expression and are
evolving as clinical predictors for diagnosis and progression of human
disease.

5. AIM
Finding correlation between increasing fibrosis and variables like
PTEN, miR-21, HCV RNA levels, HCV genotype and other biochemical
parameters
To draw a prediction model out of variables significantly affecting the
dependable variables.

6. METHODS
Study consisted of 84 CHC patients whose miR-21, PTEN expression, HCV
RNA levels were estimated by Real Time QPCR.
76 Healthy blood donors were selected randomly and were evaluated for
any history of liver disease.
All the CHC patients had undergone liver biopsy and were grouped into 3
fibrosis group, viz. ‘F0’ (Fibrosis score 0), ‘FM’(Fibrosis 1& 2), ‘FH’
(Fibrosis ≥3).
Statistical analysis was carried out using SPSS version 20.

7. RESULTS
 The average age (Mean±S.D) of CHC and control group was
42.55±11.258 and 33.42±6.293 years respectively.
The mRNA expression levels of PTEN was 11.32 fold reduced in
disease group when compared to healthy (p<0.001).
The miR-21 expression levels was increased 1.5 folds in disease group
when compared to healthy group (p<0.001).
 Univariate analysis and post-hoc analysis revealed following results
 The Total Bilirubin was significantly different between ‘FM’ and
‘FH’ (0.7±0.306 and 1.13±.563; p=0.014)
miR-21 expression levels were significantly different in ‘F0’,’FM’
and ‘FH’ group. ‘FH’ group had 2.3 fold and 4.5 fold increase in
expression when compared to ‘FM’ and ‘F0’ respectively. The ‘FM’
group had 1.9 fold increased expression than the ‘F0’ group.
PTEN expression was significantly higher in ‘FH’ group than the
‘FM’ and ‘F0’ group.

8. RESULTS (CONTD..)
Multinominal logistic regression was done to find out the predictor of
increasing fibrosis with above variables and It was found that only
PTEN(2
∆Cp
) predicted FH from FM with statistical significance (B=-
1.5,p<0.0001).
Multiple regression was carried out to predict PTEN(2
∆Cp
) from T.Bil and
miR21(2
∆Cp
).
 T.Bil and miR21(2
∆Cp
) added statistically significantly to the
prediction model
Regression equation:
PTEN(2
∆Cp
)=40.63-(13.85xmi21a(2
∆Cp
))+(10.65xT.Bil)
Using this model new cut-off value of PTEN(2
∆Cp
) was calculated using
ROC.
 AUC was 0.983
cut-off level >24.22 was found to predict higher fibrosis with
sensitivity of 92.86% and Specificity of 89.29%.
It had a PPV of 82.35% and NPV of 100%

9. CONCLUSION
Decrease in PTEN mRNA expression is significantly associated
with increase in fibrosis.
PTEN expression and miR-21 expression are negatively
correlated.
Other than PTEN expression,miR-21 expression and Total
Bilirubin, non other factor correlate to fibrosis.
 New score of >24.22 of PTEN from the equation
PTEN(2
∆Cp
)=40.63-(13.85xmi21a(2
∆Cp
))+(10.65xT.Bil) can be used for
predicting higher fibrosis.