MicroRNAs as biomarkers of cervical cancer development: a literature review on miR-125b and miR-34a

MicroRNAs are non-coding RNAs with important functions in several biological processes, such as, regulation of cell cycle, immune response, inflammation, and apoptosis. In fact, deregulation and abnormal expression of these molecules is associated with human pathologies including cancer and several have already emerged as potential prognostic biomarkers in different neoplasias. miR-34a is directly regulated by p53 and acts as tumor suppressor while miR-125b plays a significant role in immune response and apoptosis. In cervical carcinogenesis, HPV proteins seem to interact with both miR-34a and miR-125b changing its expression and promoting persistent infection and cervical cancer development. In this review we describe the potential role of miR-125b and miR-34a in cervical carcinogenesis, including interaction with HPV and mechanism of deregulation. Additionally, their clinical applications in cervical cancer as prognostic/predictive biomarkers are also briefly discussed.     

Good agreements between self and clinician-collected specimens for the detection of human papillomavirus in Brazilian patients

Women infected with human papillomavirus (HPV) are at a higher risk of developing cervical lesions. In the current study, self and clinician-collected vaginal and cervical samples from women were processed to detect HPV DNA using polymerase chain reaction (PCR) with PGMY09/11 primers. HPV genotypes were determined using type-specific PCR. HPV DNA detection showed good concordance between self and clinician-collected samples (84.6%; kappa = 0.72). HPV infection was found in 30% women and genotyping was more concordant among high-risk HPV (HR-HPV) than low-risk HPV (HR-HPV). HPV16 was the most frequently detected among the HR-HPV types. LR-HPV was detected at a higher frequency in self-collected; however, HR-HPV types were more frequently identified in clinician-collected samples than in self-collected samples. HPV infections of multiple types were detected in 20.5% of clinician-collected samples and 15.5% of self-collected samples. In this study, we demonstrated that the HPV DNA detection rate in self-collected samples has good agreement with that of clinician-collected samples. Self-collected sampling, as a primary prevention strategy in countries with few resources, could be effective for identifying cases of HR-HPV, being more acceptable. The use of this method would enhance the coverage of screening programs for cervical cancer.     

高危HPV 感染及hTERC 基因在宫颈癌发生发展中的相关性

目的:探讨高危人乳头瘤病毒及人类染色体端粒酶基因(hTERC)在宫颈癌发生发展中的关系。方法:2010年10月至2011年05月就诊于哈尔滨医科大学附属第一医院妇科患者445例,采用表面等离子体谐振法(SPR)及荧光原位杂交法(FISH)分别检测高危HPV和hTERC的表达情况,对任一结果阳性者行阴道镜下宫颈活检和病理学检查,其中炎症对照组45例,CIN组33例及宫颈癌组6例。结果:单一型HPV29例,多重型26例,最常见为HPV16型合并的二型感染(17例)。在炎症组,CINⅠ组,CINⅡ组、CINⅢ组及宫颈癌组中多重高危HPV比率为17.78%、28.57%、44.44%、50.00%、66.67%,与炎症对照组比较其他组均有统计学意义(P<0.05)。hTERC基因在各组中的阳性表达率分别为2.22%、14.29%、61.11%、75.00%及100%。CIN组及宫颈癌的hTERC基因表达与炎症组比较差异均有统计学意义(P均<0.01),其中CINⅡ及以上各组差异明显。多重HPV感染与hTERC表达呈中等正相关(r=0.462,P<0.01)。结论:随着宫颈病理类型分级升高,HPV感染及hTERC基因表达率升高,hTERC基因在宫颈癌发生发展中起重要作用;高危HPV感染与hTERC基因的异常扩增关系密切,提示二者可能存在因果关系。     

The point mutation analysis of Cyp2C9*2 (Arg144Cys C>T), Cyp2C9*3 (Ile359Leu A>C) and VKORC1 (1639G>A) in women with cervical cancer related to HPV: A case-control study

Human papillomavirus (HPV) is the most common sexually transmitted viral infection worldwide. HPV tumorigenesis genotypes are the causative agents of cervical cancer and genital malignancies. The scientific literature has demonstrated that life style, environmental, epigenetic accompanied with HR-HPV genotypes are potential risk factors for cervical cancer progression. The frequencies of the Cyp2C9*², Cyp2C9*³, and vitamin K epoxide reductase complex subunit 1 (VKORC1) genotypes as potential molecular biomarkers have been investigated on Iranian women with cervical malignancy related to HPV genotypes. As a case-control study, the mutations were appraised using a polymerase chain reaction-restriction fragment length polymorphism procedure on women suffering from HPV infection (60 cases), CC (46 cases), and 40 subjects of as healthy control. The outcomes demonstrated that Cyp2C9*³ showed a meaningful relationship between women diagnosed with cervical cancer and the healthy population (AA vs. AC; OR, 7.15; 95% CI, 1.94-26.3; p = .003). It was also observed that the Cyp2C9*³ mutation in women with cervical cancer and VKORC1 in healthy population with HPV (+), did not follow the Hardy–Weinberg equilibrium. Our findings aid understanding the genetic polymorphism distribution of Cyp2C9*², Cyp2C9*³, and VKORC1 in women with genital malignancies. This can also be useful in predicting the susceptibility risk factors for developing cervical cancer. However, allelic discrimination as a molecular biomarker requires further research.     

Modulation of apoptosis by early human papillomavirus proteins in cervical cancer

Cervical cancer (CC) constitutes a major women health problem. Clinical, molecular, and epidemiological investigations have identified persistent infection with high risk human papillomavirus (HR-HPV) as the major cause of CC. HR-HPVs lead to development of cervical carcinoma, predominantly through the action of E5, E6 and E7 viral oncoproteins. After HR-HPV infection, viral proteins employ strategies to modulate apoptosis. The E2 viral protein induces apoptosis in both normal and HPV-transformed cells through activation of caspase-8. The E5 protein can impair CD95L- and TRAIL-mediated apoptosis, which suggests that it may prevent apoptosis at early stages of viral infection. E6 inhibits apoptosis through the proteolytic inactivation of pro-apoptotic proteins such as p53, FADD, or procaspase-8, employing the ubiquitin proteasome pathway, or through interactions with proteins that form the death-inducing signaling complex (DISC) such as TNF-R1. On the other hand, E7 oncoprotein expressing cells are usually predisposed to undergo apoptosis. Useful targets for therapeutic strategies would interfere with expression or function of HR-HPV proteins to eliminate cells that express viral oncoproteins. In this review, we summarize the available data on the interaction of early HPV proteins with cellular factors that promote cell death, and the functional consequences of these interactions on apoptosis.     

A retrospective analysis of human papillomavirus (HPV) prevalence and genotype distribution among 25,238 women in Shanghai,China revealed the limitations of current HPV-based screening and HPV vaccine

BackgroundTo determine the human papillomavirus (HPV) type-specific prevalence and distribution among women with various age and cervical lesions in Shanghai, China. And to evaluate the carcinogenicity of different high-risk HPV (HR-HPV) and the efficacy of HR-HPV testing and HPV vaccine.MethodsThe clinical data from 25,238 participants who received HR-HPV testing (HPV GenoArray test kit, HybriBio Ltd) at the Affiliated Hospital of Tongji University from 2016 to 2019 were reviewed and analyzed using SPSS (version 20.0, Tongji University, China).ResultsThe overall prevalence of HPV was 45.57% in the study population, of which 93.51% were found HR-HPV infection. The three most prevalent HR-HPV genotypes were HPV 52 (22.47%), 16 (16.4%) and 58 (15.93%) among HPV-positive women, and HPV 16 (43.30%), 18 (9.28%) and 58 (7.22%) in women with histologically confirmed cervical cancer (CC). 8.25% of CC were found to be HPV negative. Only 83.51% of CC cases were related to the HPV genotypes covered by nine-valent HPV vaccine. HPV prevalence and genotype distribution varied with age and cervical histology. The odds ratios (OR) of HR-HPV for CC were also different, among which the top three types were HPV 45 [OR= 40.13, 95% confidence intervals (CI) 10.37–155.38], 16 (OR=33.98, 95%CI 15.90–72.60) and 18 (OR=21.11, 95%CI 8.09–55.09). The increase in the types of HPV infection did not increase the risk of CC correspondingly. As the primary cervical screening method, HR-HPV testing showed the high sensitivity (93.97%, 95%CI 92.00–95.49) but low specificity (42.82%, 95%CI 41.81–43.84).ConclusionsOur study provide the comprehensive epidemiological data on HPV prevalence and genotype distribution among Shanghai women with various cervical histology, which can not only serve as a significant reference for clinical practice, but also implicated the need of more effective CC screening methods and HPV vaccine covering more subtypes.     

Role of proteomics in translational research in cervical cancer

Cervical cancer is one of the leading causes of cancer morbidity and mortality in women worldwide. More than 98% of cases are related to a human papillomavirus (HPV) infection. Infection with specific subtypes of HPV has been strongly implicated in cervical carcinogenesis. The identification and functional verification of host proteins associated with HPV E6 and E7 oncoproteins may provide useful information for understanding cervical carcinogenesis and the development of cervical cancer-specific markers. In addition, proteomic profiling of altered proteins by anticancer drugs on cervical cancer cells may contribute to providing the fundamental resources for investigation of disease-specific target proteins, elucidation of the novel mechanisms of action and development of new drugs. The advent of proteomics has provided the hope of discovering novel biological markers for use in the screening, early diagnosis and prediction of response to therapy. This review describes the studies where profiles of protein expression in cervical cancer have been generated.     

Acquired immune response to oncogenic human papillomavirus associated with prophylactic cervical cancer vaccines

Human papillomavirus (HPV) is a common infection among women and a necessary cause of cervical cancer. Oncogenic HPV types infecting the anogenital tract have the potential to induce natural immunity, but at present we do not clearly understand the natural history of infection in humans and the mechanisms by which the virus can evade the host immune response. Natural acquired immune responses against HPV may be involved in the clearance of infection, but persistent infection with oncogenic virus types leads to the development of precancerous lesions and cancer. B cell responses are important for viral neutralization, but antibody responses in patients with cervical cancer are poor. Prophylactic vaccines targeting oncogenic virus types associated with cervical cancer have the potential to prevent up to 80% of cervical cancers by targeting HPV types 16 and 18. Clinical data show that prophylactic vaccines are effective in inducing antibody responses and in preventing persistent infection with HPV, as well as the subsequent development of high-grade cervical intraepithelial neoplasia. This article reviews the known data regarding natural immune responses to HPV and those developed by prophylactic vaccination.     

Usefulness of HPV testing in the follow-up of untreated cervical low grade lesions

The aim of the present work was to evaluate the usefulness of high-risk human papillomavirus (HR-HPV) testing for the follow-up of women with untreated low grade cervical squamous cell lesions (LSIL). For that, 412 women with a cytological diagnosis of LSIL at entry were monitored by cytology, HR-HPV testing with the Hybrid Capture II assay (HC-II) and colposcopy. Our primary endpoint was clinical progression defined by the presence of a high grade cervical intraepithelial neoplasia (CIN2 and CIN3) at the biopsy. At baseline, histological control revealed 10 CIN2 and 11 CIN3 only in the cohort of women HR-HPV+. In the follow-up, 4 CIN2 and 8 CIN3 were detected, always in the women initially HR-HPV+. Thus, the recurrence of a HR-HPV+ infection clearly selects a population at high-risk for CIN2-3. The semi-quantitative appreciation of the viral load with HC-II could not be used as a good prognostic factor for the follow-up of women with LSIL. HR-HPV testing reduces the number of cytology and colposcopy examinations in the follow-up of women aged >35 years when HPV testing is initially negative. Thus HR-HPV testing should be reserved for the follow-up of this population of women initially HR-HPV+ and proposed 6 to 12 months after the cytological diagnosis of LSIL.     

Gene discovery in cervical cancer : towards diagnostic and therapeutic biomarkers

Cervical cancer is a potentially preventable disease; however, it remains the second most common malignancy in women worldwide. The human papillomavirus (HPV) is the single most important etiological agent in cervical cancer. HPV contributes to neoplastic progression through the action of two viral oncoproteins E6 and E7, which interfere with critical cell cycle pathways, tumor protein p53, and retinoblastoma protein. However, evidence suggests that HPV infection alone is insufficient to induce malignant changes, and other host genetic variations are important in the development of cervical cancer. Advances in molecular biology and high throughput technologies have heralded a new era in biomarker discovery and identification of molecular targets related to carcinogenesis. These advancements have improved our understanding of carcinogenesis and will facilitate screening, early detection, management, and personalized targeted therapy. A number of these developments and molecular targets associated with cervical cancer will be addressed in this review.     

鄂中地区8136例妇女HPV感染导流杂交法亚型检测的临床意义

目的分析鄂中地区人群HPV感染基因型别。方法采集8136例鄂中地区2009年8月至2010年7月武汉市商业职工医院门诊与住院患者宫颈口及颈管脱落上皮细胞,应用导流杂交法进行HPV分型检测。结果HPV阳性感染1437例（17．66％）,其中高危型感染检出1189例（占82．67％）,低危型感染检出141例（占9．74％）,中国人群常见亚型感染检出305例（占21．16％）,多重感染检出369例（26．68％）。单一感染者中高危型833人（57．97％）,低危型感染者72人（5．01％）,中国人群常见亚型感染者163人（11．34％）。根据年龄分层,〈25岁组HPV感染率相对较高,为21．17％（P〈0．05）;HPV高危型感染组中〈25岁比例较高,达17．80％。在1437例HPV阳性感染者中,单一感染者共1068例（74．32％）,二重感染占18．72％。最常见的交叉感染是高危型＋中国人群常见亚型合并感染（8．28％）。结论导流杂交法HPV基因型分型检测可为宫颈疾病流行病学及早筛早治提供重要线索,对于发现HPV感染的高危人群、积极控制HPV感染、具有重要意义。     

Chlamydia trachomatis infection and human papillomavirus in women with cervical neoplasia in Pernambuco-Brazil

Chlamydia trachomatis (CT) is the most common bacterial cause of sexually transmitted disease. High-risk human papillomavirus (HR-HPV) is considered the main etiological agent for cervical neoplasia. Evidences showed that the presence of co-infection of CT and HR-HPV plays a central role in the etiology of cervical intraepithelial neoplasia (CIN) and cervical cancer. The goals of this study were: evaluate the human papillomavirus (HPV) and CT prevalence among Brazilian women with abnormal cytology and provide the effect of this association on the severity of cervical neoplasia. The population of this study was composed by 142 women with incident histological incidence of CIN grades I, II, III or cervical cancer from Recife, Northeast of Brazil. The polymerase chain reaction method on a cervical brush specimen was used to detect both agents and the automatic sequencing method was used for HPV genotyping assay. The prevalence of HPV and CT was 100 and 24.65 %, respectively. Thirteen types of HPV were detected; HPV 16, 18, 31 and 33 were the most common. The most prevalent HPV types were HPV 16 and 18. A significant association between CT positive and HPV 16 infection was found (p < 0.0106; OR = 5.31; 95 % IC 1.59–17.67). In the study population, there was diversity of HPV infections, with high-risk types being the most common. Also, the data collected suggest that CT infection may play an important role in the natural history of HPV infection.     

Changes in type‐specific human papillomavirus load predict progression to cervical cancer

Persistent high-risk human papillomavirus (HPV) infection is strongly associated with the development of high-grade cervical intraepithelial neoplasia or cancer (CIN3+). However, HPV infection is common and usually transient. Viral load measured at a single time-point is a poor predictor of the natural history of HPV infection. The profile of viral load evolution over time could distinguish HPV infections with carcinogenic potential from infections that regress. A case-cohort natural history study was set-up using a Belgian laboratory database processing more than 100,000 liquid cytology specimens annually. All cytology leftovers were submitted to real-time PCR testing identifying E6/E7 genes of 17 HPV types, with viral load expressed as HPV copies/cell. Samples from untreated women who developed CIN3+ (n = 138) and women with transient HPV infection (n = 601) who contributed at least three viral load measurements were studied. Only single-type HPV infections were selected. The changes in viral load over time were assessed by the linear regression slope for the productive and/or clearing phase of infection in women developing CIN3+ and women with transient infection respectively. Transient HPV infections generated similar increasing (0.21 copies/cell/day) and decreasing (−0.28 copies/cell/day) viral load slopes. In HPV infections leading to CIN3+, the viral load increased almost linearly with a slope of 0.0028 copies/cell/day. Difference in slopes between transient infections and infections leading to CIN3+ was highly significant (P < .0001). Serial type-specific viral load measurements predict the natural history of HPV infections and could be used to triage women in HPV-based cervical cancer screening.     

Population-based study on the prevalence of and risk factors for human papillomavirus infection in Qujing of Yunnan province, Southwest China

ABSTRACT: BACKGROUND: Human papillomavirus (HPV) infection causes cervical cancer and premalignant lesions of the cervix. Prevalence of HPV infection and HPV genotypes vary among different regions. However there is no data on the prevalence of HPV infection and HPV genotypes from southwest China. This study was undertaken to determine the prevalence of and risk factors for HR-HPV infection in Qujing of Yunnan province, southwest China to provide comprehensive baseline data for future screening strategies. Methods: A sample of 5936 women was chosen by the multi-stage stratified cluster sampling method with selection probabilities proportional to size (PPS). An epidemiological questionnaire was conducted via a face-to-face interview and cervical specimens were taken for HPV DNA testing by Digene Hybrid Capture 2 (HC2) test. HPV Genotyping Reverse Hybridization Test was used for HPV genotyping. Proportions were compared by Chi-squared tests, and logistic regression was utilized to evaluate risk factors. Results: The median age was 38 years and the inter-quartile range was from 31 years to 47 years. 97.3% of the study population was Han nationality. Overall prevalence of HR-HPV infection was 8.3% (494/5936) and bimodal age distribution of HPV infection was observed. The five most prevalent HR-HPV genotypes were HPV-16(3.4%), HPV-56(1.7%), HPV-58(1.4%), HPV-33(1.2%) and HPV-52(0.88%). Multiple HPV infections were identified in 50.5% (208/412) of the positive genotyping specimens. Multivariate logistic regression model indicated that parity (OR=1.35, 95% CI: 1.18-1.53, p<0.0001) was a risk factor for HR-HPV infection, and age of 50-65 years (OR=0.60, 95% CI: 0.45-0.80, p=0.0005), being married or in stable relationship (OR=0.55, 95% CI: 0.31-0.96, p=0.035) were protective factors. Conclusions: This study provided baseline data on HR-HPV prevalence in the general female population in Qujing of Yunnan province, southwest China. The finding of multiple HPV infections and bimodal age distribution revealed that HPV screening is necessary for perimenopausal women in future.     

Human papillomavirus (HPV) infection in Southern Africa: prevalence,immunity, and vaccine prospects

Human papillomavirus (HPV) associated cancers are more prevalent in developing countries compared to developed countries. The major cancer caused by HPV is cervical cancer. The humoral immune response to HPV can be a marker of past infection but may also reflect persistent infection and cervical disease. IgA antibodies to HPV in oral fluid were also found to be markers of cervical disease. Cell mediated immunity is important in clearing HPV infection and for regression of the associated lesions: this means that women infected with HIV have a high prevalence of co-infection with HPV. Good cervical screening programmes can control HPV associated cervical neoplasia. However, in countries such as South Africa, where these programmes are inadequate, there is a need for an HPV vaccine. The development of HPV vaccines is reviewed. There is a call for an inexpensive vaccine that will be accessible to the women that do not have access to adequate screening programmes and are therefore at the greatest risk of cervical cancer.     

人乳头状瘤病毒分型及病毒载量与宫颈病变的研究进展

宫颈癌作为目前妇女最常见的恶性肿瘤,新发病例在全球范围内仍处于上升趋势。已有实验证明,HPV感染是宫颈癌发生发展的必要条件。然而不同分型及其病毒载量高低仍会影响诊断率,从而影响病情,使之发展为持续性感染,最终演变为癌前病变或宫颈癌。目前,研究者对于高危型HPV病毒载量与宫颈病变之间是否存在依存关系尚存争议。研究表明,高病毒载量可能成为预测宫颈癌前病变进展的指标,然而对于病理诊断明确的宫颈癌患者而言,病毒载量并不能代表病理分期。本文综述了人乳头状瘤病毒,不同基因分型,病毒载量等因素对于宫颈病变及宫颈癌的相关性的近年研究进展,希望有助于指导临床工作。     

Prevalence of high-risk human papillomavirus cervical infection in female kidney graft recipients: an observational study

ABSTRACT: BACKGROUND: Immunosuppressive therapy protects the transplanted organ but predisposes the recipient to chronic infections and malignancies. Transplant patients are at risk of cervical intraepithelial neoplasia (CIN) and cervical cancer resulting from an impaired immune response in the case of primary infection or of reactivation of a latent infection with human papillomavirus of high oncogenic potential (HR-HPV). METHODS: The aim of this study was to assess the prevalence of HR-HPV cervical infections and CIN in 60 female kidney graft recipients of reproductive age in comparison to that in healthy controls. Cervical swabs were analyzed for the presence of HR-HPV DNA. HR-HPV-positive women remained under strict observation and were re-examined after 24 months for the presence of transforming HR-HPV infection by testing for HR-HPV E6/E7 mRNA. All the HR-HPV-positive patients were scheduled for further diagnostic tests including exfoliative cytology, colposcopy and cervical biopsy. RESULTS: The prevalence of HR-HPV did not differ significantly between the study group and the healthy controls (18% vs 25%, p=0.37). There was no correlation between HR-HPV presence and the immunosuppresive regimen, underlying disease, graft function or time interval from transplantation. A higher prevalence of HR-HPV was observed in females who had had [greater than or equal to]2 sexual partners in the past. Among HR-HPV-positive patients, two cases of CIN2+ were diagnosed in each group. In the course of follow-up, transforming HR-HPV infections were detected in two kidney recipients and in one healthy female. Histologic examination confirmed another two cases of CIN2+ developing in the cervical canal. CONCLUSIONS: Female kidney graft recipients of reproductive age are as exposed to HR-HPV infection as are healthy individuals. Tests detecting the presence of HR-HPV E6/E7 mRNA offer a novel diagnostic opportunity in those patients, especially in those cases where lesions have developed in the cervical canal.     

E-cadherin 在宫颈上皮内瘤变及宫颈癌中的表达及其与高危型HPV感染的相关性

目的:研究各级宫颈上皮内瘤变(CIN)及宫颈癌组织中E-cadherin的表达及其与高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)感染的相关性探讨其在宫颈疾病发生、发展中的意义。方法:选取2008年1月至2014年12月我院收治的150例患者标本并将其分为CINⅠ级组、CINⅡ-Ⅲ级组及宫颈癌组用免疫组化法对E-cadherin的表达情况进行检测并于术前采用PCR-反向点杂交法检测患者高危型HPV感染情况所得结果:进行统计学分析。结果:(1)E-cadherin在CINⅠ级、CINⅡ-Ⅲ级及宫颈癌中的阳性表达率分别为40/50(80.0%),24/50(48.0%)、17/50(34.0%),随疾病的进展E-cadherin的表达明显减少,各组间差异有统计学意义(P0.05)。(2)高危型HPV在CINI级、CINⅡ-Ⅲ级及宫颈癌中的阳性感染率分别为21/50(42.0%)、38/50(76.0%),48/50(96.0%),各组间差异有统计学意义(P0.05)。(3)在CIN和宫颈癌中,HR-HPV阳性组中E-cadherin阳性率39.3%(42/107)低于HR-HPV阴性组中E-cadherin阳性率90.7%(39/43)(P0.05)。结论:E-cadherin的表达下降或缺失可能是HR-HPV导致宫颈癌发生、发展的机制之一。