Results - cytology.com.au€¦ · N.Melnikova, N.Yarovaya, M.Zakharenko, I.Antonova, A.Senchukova, N. Babaeva, N.Bolotina, V.Bozhenko Federal State Budgetary Institution Russian Scientific

N.Melnikova, N.Yarovaya, M.Zakharenko, I.Antonova, A.Senchukova, N. Babaeva, N.Bolotina, V.Bozhenko

Federal State Budgetary Institution Russian Scientific Center of Roentgenoradiology of the Ministry of Healthcare of

the Russian Federation, 117997 Moscow, Profsoyuznaya str., 86, Russian Federation

Pre

dic

ted

class

ific

ati

on

s

Observed classifications

Percent

–

Correct

LSIL or

less

p=,47368

HSIL+

p=,52632

LSIL

or less 95,00000 1 19

HSIL+ 94,44444 17 1

Total 94,73684 18 20

The

classification

matrix

for 2

groups by

Discriminant

Function

Analysis

Objectives

The aim of this study was to count the

efficiency of liquid-based Pap test and the

possibility of differentiation HSIL+ from LSIL

or less based on the expression of the 24-gene

panel mRNA measurement by quantitative

PCR in the residual media Pap test CellPrep in

postmenopausal women.

Methods 38 postmenopausal women had subsequent

histological examination within 3 months after

the cytodiagnosis by Pap test CellPrep. mRNA

expression of the 21 genes (Ki-67, STK-15,

CCNB1, CCND1, MYC, MYBL2, P16INK4A,

PTEN, BIRC5, BCL2, BAG1, TERT, NDRG1,

ESR1, PGR, HER2, GRB7, MGB1, MMP11,

CTSL2, CD68) and 3 house-keeping genes

(GUSB, HPRT1, B2M) was measured by

quantitative PCR in the Pap test CellPrep

media. The study of this set of genes is

included in the diagnostic kit - GlobalIndex

(GI) - a registered test system for predicting

the recurrence of breast cancer.

Results The histological examination was classified in such clinical

groups: normal epithelium/benign tissue changes (16 patients),

LSIL (3 patients), HSIL (16 patients), squamous cell carcinoma (2

patients), endocervical adenocarcinoma (1 patient). We observed

that 3 cases of LSIL and 1 case of HSIL were false positive and the

positive predictive value for the CellPrep Pap test was 80.95%

95% (CI 65.70% to 90.41%.) The accuracy of the Pap smear was

80% (95% CI 61.43% to 92.29%.) . We discovered that combined

evaluation of the 24 gene expression panel allows, according to the

discriminant analysis, to carry out the correct differentiation for

HSIL+ from (LSIL or less) in 94,74% of cases.

Conclusions

The inclusion of the expression analysis of

24-gene panel by quantitative PCR to the Pap

test CellPrep could lead to more accurate

detecting for severe cervical lesions

The correct classification is mainly influenced

by the estimation of the expression of 6 genes:

MYBL2,HER2,NDRG1,BAG1,GRB7, BCL2.

The estimation of levels ESR1 and MYBL2

mRNA expression has a crucial meaning by

forward stepwise analysis.

2 ml media

HSIL. Histological

examination. (H&Ex400)

1-HSIL. CellPrep Pap test.

(Pap stain x400) Endocervical

adenocarcinoma. (H&Ex400)

Adenocarcinoma CellPrep

Pap test (Pap stain x400)

Median

25%-75%

Min-Max

260

240

220

200

180

160

140

120

100

80

60

40

20

0

Boxplot by Group

Variable: ESR1

ESR

1

b a

120

100

80

60

40

20

0

-20

Boxplot by Group

Variable: MYBL2

MY

BL

2

b a

Groups of histological diagnosis a – LSIL or less; b – HSIL+

The estimation of ESR1 and MYBL2 mRNA levels in groups with histological confirmed

LSIL or less (normal epithelium/benign tissue changes

of the cervix) and HSIL+

Real-time cycler DT

Prime5, Company

DNA-Technology

LLC, Russia