The Status of Human Papillomavirus and Tumor Suppressor Genesp53andp16in Carcinomas of Uterine Cervix from India

doi:10.1006/gyno.1998.4991

Gynecologic Oncology

Volume 69, Issue 3, June 1998, Pages 205-209

https://doi.org/10.1006/gyno.1998.4991 Get rights and content

Abstract

Objectives.Infection with the high-risk strain of human papillomaviruses (HPVs) and the inactivation of the tumor suppressor genep53through mutation are important factors in cervical carcinogenesis. To know whether such events would occur in cervical carcinomas of Indians, 43 tumors (consisting of 36 of stage III B and 6 of stage II B) were screened forp53andp16gene mutations.

Methods.PCR followed by single-strand conformation polymorphism (SSCP) analysis were used to detect mutations inp53andp16genes and PCR for the presence of human papillomavirus genome. HPV status was ascertained by PCR amplification of parts of E6 and E7 genes using primers pU-1M and pU-2R and typing was carried out by restriction analysis.

Results.Of the 43 samples analyzed, 4 samples (9%) showed mobility shifts forp53mutations; PCR products of thep16gene did not show band shifts in SSCP analysis. HPV DNA was detected in 70% of the 43 samples analyzed: HPV 16 in 23 cases (53%), HPV 18 in 4 cases (13.3%), and HPV 33 in 1 case (3.3%). Two amplified HPV DNAs that were difficult to type with various restriction enzymes were cloned and the amplified regions were sequenced. One of these was 93% close to HPV 35 and the other was 80% close to HPV 58. Three samples had bothp53mutations and HPV genome.

Conclusions.Our results indicate that HPV 16 infection was more common than HPV 18, thep53mutations and HPV infection were not mutually exclusive events in the genesis of carcinoma of uterine cervix among Indian women, andp16gene may not play a role in Indian cervical carcinomas.

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Cited by (34)

p16<sup>INK4a</sup> immunostaining in cytological and histological specimens from the uterine cervix: A systematic review and meta-analysis
2009, Cancer Treatment Reviews
Citation Excerpt :
Amongst the relevant studies, thirty-six failed to meet the inclusion criteria and were excluded from this overview. Nine of the studies assessed p16 immunostaining in glandular20–28 and twenty-one in invasive cervical lesions only.29–49 Another six studies represented duplicate reports and were subsequently excluded50–55 (Fig. 1).
P16^INK4a is a biomarker for transforming HPV infections that could act as an adjunct to current cytological and histological assessment of cervical smears and biopsies, allowing the identification of those women with ambiguous results that require referral to colposcopy and potentially treatment.
We conducted a systematic review of all studies that evaluated the use of p16^INK4a in cytological or histological specimens from the uterine cervix. We also estimated the mean proportion of samples that were positive for p16^INK4a in cytology and histology, stratified by the grade of the lesion.
Sixty-one studies were included. The proportion of cervical smears overexpressing p16^INK4a increased with the severity of cytological abnormality. Among normal smears, only 12% (95% CI: 7–17%) were positive for the biomarker compared to 45% of ASCUS and LSIL (95% CI: 35–54% and 37–57%, respectively) and 89% of HSIL smears (95% CI: 84–95%). Similarly, in histology only 2% of normal biopsies (95% CI: 0.4–30%) and 38% of CIN1 (95% CI: 23–53%) showed diffuse staining for p16^INK4a compared to 68% of CIN2 (95% CI: 44–92%) and 82% of CIN3 (95% CI: 72–92%).
Although there is good evidence that p16^INK4a immunostaining correlates with the severity of cytological/histological abnormalities, the reproducibility is limited due to insufficiently standardized interpretation of the immunostaining. Therefore, a consensus needs to be reached regarding the evaluation of p16^INK4a staining and the biomarker needs to be assessed in various clinical settings addressing specific clinical questions.
Human Papillomavirus Infection and Cervical Cancer Prevention in India, Bangladesh, Sri Lanka and Nepal
2008, Vaccine
Citation Excerpt :
Determining the age that accurately reflects first exposure will be critical to the formulation of a coherent HPV vaccination strategy in India. The prevalence of HPV types in cervical cancer specimens, analyzed by PCR or HC2, varied between 75–99% in hospital-based cross-sectional studies in different regions of India (Table 4) [11,12,14,17–20]. HPV-16 or 18 were the most common in all regions with some difference in the HPV types subsequently, but no substantial difference in the type attributable fraction.
Although one-third of the world cervical cancer burden is endured in India, Bangladesh, Nepal and Sri Lanka, there are important gaps in our knowledge of the distribution and determinants of the disease in addition to inadequate investments in screening, diagnosis and treatment in these countries. Prevalence of human papillomavirus (HPV) infection among the general populations varies from 7–14% and the age-specific prevalence across age groups is constant with no clear peak in young women. This observation may be the result of a low clearance rate of incident infections, frequent re-infection/reactivation, limited or no data in target high-risk age groups (teenagers), and sexual behavioural patterns in the population. High-risk HPV types were found in 97% of cervical cancers, and HPV-16 and 18 were found in 80% of cancers in India. Beyond research studies, demonstration projects and provincial efforts in selected districts, there are no serious initiatives to introduce population-based screening by public health authorities in these countries. Cervical cancer is a relatively neglected disease in terms of advocacy, screening and prevention from professional or public health organizations. Cytology, HPV testing and visual screening with acetic acid (VIA) or Lugol's iodine (VILI) are known to be accurate and effective methods to detect cervical cancer and could contribute to the reduction of disease in these countries. While HPV vaccination provides hope for the future, several barriers prohibit the introduction of prophylactic vaccines in these countries such as high costs and low public awareness of cervical cancer. Efforts to implement screening based on the research experiences in the region offer the only currently viable means of rapidly reducing the heavy burden of disease.
A meta-analysis of human papillomavirus type-distribution in women from South Asia: Implications for vaccination
2008, Vaccine
Citation Excerpt :
All statistical analyses were done using Stata 9.0 statistical package. Of the studies from India, Pakistan and Sri Lanka, nine studies from India fulfilled the inclusion criteria [13–21]. Table 1 shows the details of these with respect to numbers examined, region studied, PCR primers, etc.
To determine human papillomavirus (HPV) prevalence and type-distribution in women from South Asia, with and without cervical lesions, in order to estimate the impact of an HPV 16/18 prophylactic vaccine in this region and to assess additional types that should be incorporated in new vaccines.
A meta-analysis was conducted that included studies using polymerase chain reaction to detect HPV-16, -18, -6, -11 and at least one other HPV type, with a minimum of 20 cases in each grade of lesion. Total as well as type-specific prevalence of various HPV types were estimated, stratified by cervical lesion grade, using Stata 9.0 software package.
Nine studies from India fulfilled the inclusion criteria. A total of 558, 52, 52 and 3061 women, respectively with invasive cervical cancer (ICC), high-grade squamous intraepithelial lesions (HSIL), low-grade squamous intraepithelial lesions (LSIL) and normal cytology/histology were included. Overall HPV prevalence was 94.6%, 86.5%, 65.4% and 12.0% in women with ICC, HSIL, LSIL and normal cytology/histology, respectively. In ICC, HPV-16 was the predominant type (64.8%), followed by HPV-18, -45, -33, -35, -58, -59 and -31. The estimated HPV-16/18 positive fraction was 78.9% in women with ICC (87.7% in North and 77.2% in South India), 61.5% with HSIL, 30.8% with LSIL and 3.9% in women with normal cytology/histology. There was no difference in overall HPV prevalence in cervical cancer between North and South India (P = 0.063). However, HPV-16 and -45 appeared to be more prevalent in North India (P = 0.018 and 0.013, respectively), while HPV-35 appeared to be more prevalent in South India (P = 0.033).
It is estimated that HPV-16/18 vaccines will provide over 75% protection against ICC in South Asia. HPV-45, -33, -35 and -58 account for an additional 20% of cervical cancer in this region. The addition of these additional HPV types in a second-generation vaccine could provide optimal cervical cancer prevention in this region.
HPV16/18 prevalence in cervical lesions/cancers and p53 genotypes in cervical cancer patients from India
2002, Gynecologic Oncology
Objectives. The HPV16/18 code for an oncoprotein-E6, which binds to p53 tumor suppressor protein and degrades the protein via ubiquitination. A common polymorphism of p53 in exon 4 codon 72, resulting in either proline (Pro) or arginine (Arg), affects HPV16/18 E6-mediated degradation of p53 protein in vivo. Hence, in the current study we investigated the prevalence of HPV16/18 in cervical lesions and the distribution of p53 genotypes in cervical cancers and normal healthy women.
Methods. DNA from 337 Indian women with invasive cervical cancers, 164 women with clinically normal cervix, 64 women with low-grade squamous intraepithelial lesions (LSIL), and 5 women with high-grade squamous intraepithelial lesions (HSIL) was examined for the presence of HPV16/18 using consensus primers in a polymerase chain reaction (PCR), and the specific HPV type was identified by Southern hybridization of the PCR product using HPV16/18 type-specific nucleotide sequences as probes. Further, 134 women with cervical cancers and 131 healthy women were used to determine the frequency of p53 genotypes, Pro/Pro, Arg/Arg, and Pro/Arg, using peripheral blood cell DNA to indicate the constitutional genotypes and allele-specific primers, in a PCR-based assay.
Results. We observed a prevalence of HPV16/18 in 77% (258/337) of cervical cancer patients, 38% (24/64) of LSILs, 4 of 5 HSILs, and 15.2% (25/164) of normal healthy women. The frequency of distribution of the three genotypes of p53 codon 72 in a subgroup of the HPV16/18-positive cervical cancer patients was Pro/Pro 0.18 and Arg/Arg 0.26, with the heterozygous Pro/Arg 0.56, differing significantly from the genotype frequency in the normal healthy women (χ² = 6.928, df = 2, P < 0.05).
Conclusions. A high prevalence of HPV16/18 was observed in the cervical cancers. The prevalence in LSILs confirms HPV16/18 infection as an early event and further indicates a role in progression of lesions. The p53 genotype distribution indicated that women homozygous for Arg genotype were at a 2.4-fold higher risk for developing HPV16/18-associated cervical carcinomas, compared to those showing heterozygous Pro/Arg genotype (odds ratio 2.4, 95% confidence interval 1.89 to 3.04).
p53 immunohistochemical expression: Messages in cervical carcinogenesis
2002, Pathology
The pattern of p53 expression was studied in pre-invasive and invasive cervical carcinoma in an attempt to clarify its role in cervical carcinogenesis.
A total of 234 invasive cervical carcinomas (152 squamous cell carcinomas, 61 adenocarcinomas and 21 adenosquamous carcinomas) and 16 cervical intraepithelial neoplasia (CIN) I, six CIN II and 25 CIN III were immunohis-tochemically studied for p53.
p53 was detected more frequently in CIN and invasive carcinoma (100% of CIN I, 74.2% CIN II + III and 70.1% invasive carcinoma) compared with benign cervices (P <0.001); however, only three squamous cell carcinomas, 11 adenocarcinomas and two adenosquamous carcinomas exhibited p53 expression in > 75% of tumour nuclei. Six of the 11 adenocarcinomas and both adenosquamous carcinomas were poorly differentiated compared with one of the three squamous carcinomas. p53 immunoreactive cells were randomly distributed in invasive carcinoma, confined to the lower third of the epithelium in CIN I, reached the middle third in 20% of CIN II and upper third in 16.6% of CIN III.
Assuming that p53 immunoreactivity indicates gene mutation when the majority (>75%) of neoplastic cells express p53, p53 mutations would seem uncommon in cervical carcinogenesis. Nonetheless, glandular malignancies, in particular poorly differentiated variants, may show a higher frequency of mutation. p53 was detected more frequently in CIN I compared with CIN II/III and invasive carcinoma which may be due to p53 protein degradation following interaction with high risk human papillomavirus E6 protein in CIN II/III and invasive carcinoma.
Molecular genetic study to detect prevalence of high-risk human papilloma virus strains (type 16 and 18) in cervical lesions and asymptomatic healthy subjects of rural central India
2019, Journal of Cytology

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Regular ArticleThe Status of Human Papillomavirus and Tumor Suppressor Genesp53andp16in Carcinomas of Uterine Cervix from India

Abstract

Virology

Lancet

Lancet

Biochim Biophys Acta

Human papilloma virus and invasive cervical carcinoma in Brazil

Br J Cancer

Human papilloma virus infection of the cervix: relative risk associations of 15 common anogenital types

Obstet Gynaecol

Papilloma virus DNA from a cervical carcinoma and its prevalence in cancer biopsy samples from different geographic regions

Proc Natl Acad Sci USA

Papilloma virus infections—a major cause of human cancers

Biochim Biophys Acta

Association of human papilloma virus types 16 and 18 E6 proteins withp53.

Science

The human papilloma virus-16 E7 oncoprotein is able to bind to the retinoblastoma gene product

Science

The state of thep53

Am J Hum Genet

The role ofp53

Br J Cancer

A new regulatory motif in cell-cycle control causing specific inhibition of cyclinD/cdk4

Nature

Germline 16 mutations in familial melanoma

Nat Genet

Alterations of thep16INK4Agene in human ovarian cancers

Mol Carcinogen

Frequent somatic mutation of theMTS1/CDK41

Cancer Res

Molecular Cloning: A Laboratory Manual

Regular Article
The Status of Human Papillomavirus and Tumor Suppressor Genesp53andp16in Carcinomas of Uterine Cervix from India

Alterations of thep16^INK4Agene in human ovarian cancers