The Role of Human Telomerase Catalytic Subunit mRNA Expression in Cervical Dysplasias

2005 ◽  
Vol 230 (4) ◽  
pp. 263-270 ◽  
Author(s):  
A. Tsezou ◽  
P. Oikonomou ◽  
P. Kollia ◽  
I. Mademtzis ◽  
E. Kostopoulou ◽  
...  
Keyword(s):  
High Risk ◽  
Mrna Expression ◽  
Telomerase Activity ◽  
Hpv Infection ◽  
Htert Mrna ◽  
Hpv Dna ◽  
Human Telomerase ◽  
High Risk Hpv ◽  
Intraepithelial Lesions ◽  
Htert Mrna Expression

Telomerase activity and human telomerase reverse transcriptase (hTERT) mRNA expression were investigated in cervical specimens and were correlated with cytologic findings and the presence of human papilloma virus (HPV) infection. Telomerase activity was evaluated by the telomeric repeat protocol assay and hTERT mRNA expression was evaluated by reverse transcriptase polymerase chain reaction (PCR). HPV DNA was detected by PCR, as well as restriction endonuclease digestion. HPV DNA was detected in all 82 specimens with abnormal cytologic findings and in 4 of 34 normal samples. Low-grade squamous intraepithelial lesions (LGSILs) were present in 74 of 82 specimens (90.2%) and high-grade squamous intraepithelial lesions (HGSILs) were present in 8 of 82 (9.75%) specimens. Seven of the eight HGSIL (87.5%) and 26 of 74 LGSIL (35.1%) specimens were hTERT positive, whereas all normal specimens were hTERT mRNA negative. Telomerase activity was detected in 21 of 74 (28.4%) LGSIL/atypical squamous epithelial cells of undetermined significance (ASCUS) and in five of eight (62.5%) HGSIL samples. A correlation was observed among telomerase activity, hTERT mRNA expression, and high-risk HPV infection in HGSIL samples (P < 0.001). High-risk HPV infection assessment showed 75% sensitivity and 72.2% specificity for HGSILs. Telomerase activity assessment in cervical smears showed sensitivity and negative predictive value (NPV) for HGSILs 62.5% and 96.7%, whereas specificity and positive predictive value (PPV) were 80.5% and 19.2%, respectively, hTERT mRNA expression assessment showed 87.5% sensitivity and 98.7% NPV for HGSILs, whereas specificity and PPV were 76% and 21.2%, respectively. Based on the above-described telomerase assessment values, it is suggested that the telomerase system might not be an appropriate diagnostic marker for cytology, given that the final evaluation must rely on a combination of all available test assessment data, clinical diagnosis, as well as the follow-up of all LGSIL samples that were positive for telomerase activation.

scholarly journals Tissue Detection and Typisation of Human Papillomavirus in Women with Squamous Intraepithelial Lesions and Squamous Invasive Carcinoma of the Cervix

2015 ◽  
Vol 69 (2) ◽  
pp. 78-85
Author(s):  
Drage Dabeski ◽  
Dragan Danilovski ◽  
Vesna Antovska ◽  
Neli Basheska ◽  
Zora Popovska ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
High Risk ◽  
Invasive Carcinoma ◽  
Hpv Infection ◽  
Oncogenic Potential ◽  
Hpv Dna ◽  
Cervical Biopsy ◽  
High Risk Hpv ◽  
Intraepithelial Lesions

AbstractIntroduction. The most common risk factor for intraepithelial lesions and cervical carcinoma is infection with human papillomavirus (HPV), especially with high-risk HPV genotypes. Only persistent, high-risk HPV infections represent a major risk factor for intraepithelial lesions and cervical cancer. The aims of the study were: detection and typisation of HPV genotypes, which are the most common causes of intraepithelial lesions and cervical cancer, determination of the correlation between HPV infection and histopathological diagnosis, and the correlation between the grade of lesion of the cervix and oncogenic potential of the virus as well as determination of the most affected age group of patients.Methods. This cross-sectional study included 100 sexually active patients with an abnormal Pap test at the age from 20 to 69 years (39±10.77), and was conducted at the University Clinic of Gynecology and Obstetrics in Skopje and University Clinic of Radiotherapy and Oncology in Skopje in the period from January 2014 to August 2014. In all patients colposcopic cervical biopsy was made with endocervical curettage for histopathological analysis and cervical biopsy for detection and HPV typisation. HPV detection and typisation were done using polymerase chain reaction (PCR) and reverse hybridization.Results. HPV DNA was detected in 81.0% (81/100) of the examined women. The relationship between the prevalence of high-risk and low-risk HPV DNA genotypes was 72.0%:9.0%. The frequency of high-risk HPV DNA genotypes ranged from: 54.5% (12/22) in productive HPV infection-mild dysplasia, 86.4% (19/22) with moderate dysplasia, 91.2% (21/23) in severe dysplasia to 100% of squamous cell carcinoma in situ (6/6) and invasive squamous cell carcinoma (5/5). Mixed HPV infection was detected in 19.0% (19/100) of all patients, in 23.5% (19/81) of HPV DNA positive patients. The most common HPV DNA genotypes, in descending order, were HPV 16 (43.2%), HPV 31 (28.4%), HPV 18 (14.8%), etc. The highest frequency of HPV infection was found in patients under 30 years of age.Conclusion. There was an association between HPV infection and squamous intraepithelial lesions and squamous invasive carcinoma of the cervix. There was a correlation between the grade of cervical lesion and the oncogenic potential of the virus. The results of this study may be useful for building a national strategy in the fight against cervical cancer.

scholarly journals EGFR and hTERT Expression as a Diagnostic Approach for Non-small Cell Lung Cancer in High Risk Groups

2010 ◽  
Vol 2 ◽  
pp. BIC.S3383 ◽  
Author(s):  
Radostina Cherneva ◽  
Ognian Georgiev ◽  
Ivanka Dimova ◽  
Blaga Rukova ◽  
Danail Petrov ◽  
...  
Keyword(s):  
Gene Expression ◽  
Lung Cancer ◽  
High Risk ◽  
Small Cell Lung Cancer ◽  
Mrna Expression ◽  
Small Cell ◽  
Htert Mrna ◽  
Small Cell Lung ◽  
The Mean ◽  
Htert Mrna Expression

Objective The early detection of NSCLC is of importance because it provides chances for better outcomes. The aim of the study was to explore the clinical utility of EGFR and hTERT mRNA expression as markers for diagnosis of NSCLC. Methods EGFR and hTERT mRNA were quantified by quantative reverse transcription real time polymerase chain reaction in plasma of 45 non-small cell lung cancer (NSCLC) and 40 chronic obstructive pulmonary disease (COPD) patients, selected by certain spirometric characteristics that made them at high risk of developing lung cancer in future. Results The gene expression level of each gene was calculated and given as a relative quantity–-RQ. EGFR gene expression was found in all lung cancer patients. The mean level of expression was RQ = 29.39. hTERT mRNA could be detected in 88% of patients. The mean expression ratio in them was RQ = 17.31. Only 50% of the high risk patients turned to be positive for EGFR. The level of their expression was RQ = 2.09. The plasma levels of hTERT could be detected in 17 (42.5%) patients of the high risk COPD group. Their mean level of expression was RQ = 1.02. A statistically significant difference in EGFR and hTERT mRNA expression could be observed between the two groups of patients–-p = 0.0001. Conclusion EGFR and hTERT mRNA are potential markers for lung cancer diagnosis, whose clinical importance should be replicated in a larger cohort of patients.

scholarly journals Detection and estimation of human papillomavirus viral load in patients with cervical lesions

2014 ◽  
Vol 39 (2) ◽  
pp. 86-90 ◽  
Author(s):  
T Rahman ◽  
S Tabassum ◽  
M Jahan ◽  
A Nessa ◽  
Dr Ashrafunnessa
Keyword(s):  
Human Papillomavirus ◽  
Viral Load ◽  
High Risk ◽  
Invasive Cervical Cancer ◽  
Hpv Infection ◽  
Cervical Lesions ◽  
Hpv Dna ◽  
Promising Tool ◽  
High Risk Hpv ◽  
Hpv Viral Load

Human papillomavirus (HPV) high risk genotype infection and HPV viral load influences the development of invasive cervical cancer and cervical intra-epithelial neoplasia (CIN). HPV DNA testing for screening of cervical cancers may play a potential role in its early detection and management. The present study detected HPV DNA and estimated HPV viral load in different types of cervical lesions among Bangladeshi women. Using the Hybrid Capture 2 (HC2) assay, HPV DNA was tested among 68 women between 25-70 years of age. A total of 13 (19.1%) cases were positive for HPV DNA. The highest viral load (501 x 10³ copies/ml) was detected in a patient with invasive carcinoma, while the lowest viral load (105 x 10³ copies/ml) was detected from a case of chronic cervicitis. The mean viral load in CIN I was 119.25 x 10³±12.5 x 10³ copies/ml (range: 110 x 10³ - 137 x 10³) and 208.50 x 10³ ± 0.59 x 10³ copies/ml (range: 139 x 10³-305 x 10³) in CIN II / III. Interestingly, HPV DNA was detected from a patient with normal cytological findings. Our study observed a moderate presence of high-risk HPV genotypes among women with cervical lesions. The HPV viral load varied with the age of the patients and stage of cervical lesions. The HC2 assay is a promising tool for diagnosing high-risk HPV infection especially before cytology tests show any abnormality. DOI: http://dx.doi.org/10.3329/bmrcb.v39i2.19648 Bangladesh Med Res Counc Bull 2013; 39: 86-90

The Regulation of Telomere Binding Proteins on Telomerase during Differentiation of ATRA Responsive and Resistant NB4 Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3390-3390
Author(s):  
He Huang ◽  
Jie Sun ◽  
Yuan Yuan Zhu ◽  
Jian Ping Lan ◽  
Xiao Yu Lai
Keyword(s):  
Protein Expression ◽  
Mrna Expression ◽  
Peak Level ◽  
Telomerase Activity ◽  
Expression Level ◽  
Htert Mrna ◽  
Down Regulation ◽  
Nb4 Cells ◽  
Significant Change ◽  
Htert Mrna Expression

Abstract It has been reported that the down-regulation of telomerase activity associated with maturation of APL cells is not mechanistically linked to cell maturation, and requires only a RAR, but not a RXR-dependent pathway. However, it is not clear whether and how telomeric proteins respond to the retinoid treatment. Using maturation-sensitive and resistant APL cell lines NB4, NB4-R1 and NB4-R2 cells, we analyzed a panel of telomeric proteins using western blotting analyses in addition to temporal profile of corresponding mRNA during the course of retinoid-induced differentiation. Our analyses show hTERTmRNA expression decreased rapidly during differentiation of NB4 and NB4-R1 cells, telomerase activity also declined. But in NB4-R2 cells, hTERT mRNA was initially decreased to 38.2% on day1 (P<0.05) and then increased to 80.0% on day 3 (P<0.05). Telomerase activity remained unchanged overtime (P>0.05), which may be caused by the increasing of hTERT mRNA expression during its later period of differentiation. TRF1 mRNA and protein expression have no significant change during differentiation in NB4 and NB4-R1 cells but has a small increase in NB4-R2 cells. The TRF1 mRNA expression level has no significant change during differentiation of NB4 and NB4-R1 cell line cells. However, it was increased to 235% on day 2 (P<0.05) and remains at this level until day 3 during the differentiation of NB4-R2 cells. TRF1 protein expression level also remains stable during differentiation of NB4 and NB4-R1 cells, but has a little increase in NB4-R2 cells. This indicates TRF1 has different regulation in RARα dependent or RXRα dependent pathways. Pinx1 mRNA expression decreased during the differentiation of NB4 and NB4-R1. But during the differentiation of NB4-R2, Pinx1mRNA expressions level was initially decreased to 34.3% (P<0.05) on day 1 then increased to 64.5% (P<0.05 compared to day1) on day2. The change of Pinx1 mRNA expression and hTERT mRNA expression in NB4(r=0.902, P=0.036), NB4-R1(r=1.00, P<0.001), and NB4-R2(r=0.880, P=0.049) cells are positive correlated. Pinx1 is the only telomere binding protein that can bind to hTERT directly, it might be responsible for the different regulation of telomerase activity through RARα dependent or RXRα dependent pathways. During NB4 cell differentiation, TANK1 mRNA expression decreased gradually to 31.6% (P<0.05) on day1, and remained this level until day3. In NB4-R1 cells, TANK1 mRNA expression increased initially to 197% at 12h(P<0.05), and then decreased gradually to 111% (P<0.05) on day 3. During the differentiation of NB4-R2 cells, TANK1 mRNA expression was initially increased to 204% at 12h(P<0.01), and then decreased gradually to 96.9% on day3 (P<0.01). Its protein expression initially increased and reached a peak level at day 1 and then decreased in the later period of differentiation of all three NB4 cells. Both TANK1 mRNA expression and its protein expression were down-regulated at the later period of differentiation in all three NB4 cells. It seems that TANK1 may act as a positive regulator on telomerase activity during differentiation. TANK2 mRNA expression remained no change during differentiation of three NB4 cells. As results show, Pinx1 and TANK1 may interfere in the regulation of telomerase. The decrease of TANK1 may be the cause of the down-regulation of telomerase activity. Further studies will focus on the mechanism of their regulation on telomerase.

The Evaluation of p16ink4a Immunoexpression/Immunostaining and Human Papillomavirus DNA Test in Cervical Liquid-Based Cytological Samples

2011 ◽  
Vol 21 (1) ◽  
pp. 79-85 ◽  
Author(s):  
Maria Nasioutziki ◽  
Angelos Daniilidis ◽  
Kostos Dinas ◽  
Maria Kyrgiou ◽  
George Valasoulis ◽  
...  
Keyword(s):  
High Risk ◽  
Hpv Infection ◽  
Low Grade ◽  
Squamous Intraepithelial Lesion ◽  
Dna Test ◽  
Hpv Dna ◽  
Squamous Cells ◽  
Intraepithelial Lesion ◽  
High Risk Hpv ◽  
Hpv Dna Test

Aim:To evaluate the role of p16INK4a immunoexpression and human papillomavirus (HPV) DNA test for the detection of dyskaryotic cells in high-risk women.Materials and Methods:This work was a retrospective diagnostic study conducted in the University Hospital of Thessaloniki from January to December 2008. The subjects were women with current or previous HPV infection and current or previous cervical intraepithelial lesion (with or without treatment) or clinical warts. All liquid-based cytological samples were tested for P16INKa and HPV DNA test. The accuracy parameters used for the outcome included sensitivity, specificity, and positive predictive value.Results:A total of 226 women were included; the mean age was 29 years. Expression of p16INK4a was detected in the cytological samples of 13% of the negative cases, 44% of the cases of atypical squamous cells of undetermined significance, 46% of the cases of low-grade squamous intraepithelial lesion, and 78% of the cases of high-grade squamous intraepithelial lesion. A total of 91 women tested positive for high-risk HPV infection, and 54 of those had p16INK4a-positive staining reaction cells. The concordance between the 2 tests, HPV DNA and p16, was 59% regarding infection-positive cases. Diffuse strong parabasal p16INK4a immunostaining (nuclear score >2) was observed in 17 cases of the abnormal cytological findings (atypical squamous cells of undetermined significance, 2 cases; low-grade squamous intraepithelial lesion, 8 cases; high-grade squamous intraepithelial lesion, 7 cases). Colposcopy-directed biopsies were used as the criterion standard for the detection of cervical intraepithelial neoplasia in 91 women. The sensitivity of p16INK4a was 95% and the specificity was 92%, whereas the sensitivity of high-risk HPV was 100% and the specificity was 78%. The positive predictive value of p16INK4a was 71%, whereas that of HPV DNA was 44%.Conclusion:The findings suggest that p16INK4a immunostaining can improve the accuracy of cytological examination and HPV DNA test and may be particularly useful in the triage of low-grade lesions.

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