Discovery and validation of FBLN1 and ANT3 as potential biomarkers for early detection of cervical cancer

Abstract Background To validate markers for cervical carcinoma (CC) and precancerous lesions related with HPV infections. Methods Three different cervical cancer cell lines C-33A, SiHa and Caski were used for secretome profiling by label-free quantitative proteomics. Cervical exfoliated cells and matching serum samples were collected from 284 patients with normal control (n = 75, 26.41 %), precancerous lesions (n = 88, 30.99 %) and early stage cervical squamous carcinoma (n = 121, 42.61 %). HPV subtyping and quantification was performed by PCR and hybridization. 20 candidate proteins identified in previous screening studies (tissue, plasma, cells) were quantified by ELISA. Finally, highly quantitative parallel reaction monitoring mass spectrometry was used to assess the specificities and sensitivities of candidate serum markers. Results While CC was found to be associated with high-risk HPV subtypes, serum antibodies for high risk HPV were not significantly related to the progression of cervical cancer. Significant differences between patient groups were detected for the four proteins CLU, APOA4, APOE and MLH3, but none would allow clinical application due to insufficient sensitivity and specificity and large variability. Subsequent proteomic secretome analysis of cervical cancer cell lines identified a set of 729 common proteins. Cross referencing this dataset with ELISA measurements revealed six candidate proteins of which two, FBLN1 and ANT3, showed co-occurrence with HPV infection (75.7 % and 85 %, respectively) and had promising diagnostic ability in terms of sensitivity and specificity. After the loss of E6/E7 by using CRISPR/Cas9 gene editing, the content of ANT3 and FBLN1 in KoE6/E7 SiHa were downregulated, which indicated the expression of ANT3 and FBLN1 in cervical cancer may be affected by HPV infection. Conclusions FBLN1 and ANT3 might be potential tumor- and HPV-associated serum markers.

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The application of CRISPR/Cas9 system in cervical carcinogenesis

Cancer Gene Therapy ◽

10.1038/s41417-021-00366-w ◽

2021 ◽

Author(s):

Chun Gao ◽

Ping Wu ◽

Lan Yu ◽

Liting Liu ◽

Hong Liu ◽

...

Keyword(s):

Cervical Cancer ◽

High Risk ◽

Transgenic Mice ◽

Cell Line ◽

Cell Lines ◽

Cancer Cell ◽

Precancerous Lesions ◽

Cancer Cell Lines ◽

Cervical Carcinogenesis ◽

Cervical Precancerous Lesions

AbstractIntegration of high-risk HPV genomes into cellular chromatin has been confirmed to promote cervical carcinogenesis, with HPV16 being the most prevalent high-risk type. Herein, we evaluated the therapeutic effect of the CRISPR/Cas9 system in cervical carcinogenesis, especially for cervical precancerous lesions. In cervical cancer/pre-cancer cell lines, we transfected the HPV16 E7 targeted CRISPR/Cas9, TALEN, ZFN plasmids, respectively. Compared to previous established ZFN and TALEN systems, CRISPR/Cas9 has shown comparable efficiency and specificity in inhibiting cell growth and colony formation and inducing apoptosis in cervical cancer/pre-cancer cell lines, which seemed to be more pronounced in the S12 cell line derived from the low-grade cervical lesion. Furthermore, in xenograft formation assays, CRISPR/Cas9 inhibited tumor formation of the S12 cell line in vivo and affected the corresponding protein expression. In the K14-HPV16 transgenic mice model of HPV-driven spontaneous cervical carcinogenesis, cervical application of CRISPR/Cas9 treatment caused mutations of the E7 gene and restored the expression of RB, E2F1, and CDK2, thereby reversing the cervical carcinogenesis phenotype. In this study, we have demonstrated that CRISPR/Cas9 targeting HPV16 E7 could effectively revert the HPV-related cervical carcinogenesis in vitro, as well as in K14-HPV16 transgenic mice, which has shown great potential in clinical treatment for cervical precancerous lesions.

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Human Papillomavirus and Pap Smear

American Journal of Lifestyle Medicine ◽

10.1177/1559827611402581 ◽

2011 ◽

Vol 6 (1) ◽

pp. 31-44

Author(s):

Indra Balachandran

Keyword(s):

Cervical Cancer ◽

Human Papillomavirus ◽

High Risk ◽

Pap Smear ◽

Precancerous Lesions ◽

The United States ◽

Hpv Infection ◽

Hpv Testing ◽

Pap Smear Screening ◽

High Risk Hpv

High-risk human papillomavirus (HPV) infection and viral persistence is a major risk factor in the development of squamous intraepithelial lesions and invasive carcinoma of the cervix. In the United States, deaths due to squamous cell carcinoma of the cervix have fallen by 75% since the 1960s because of Papanicolaou (Pap) smear screening. However, the traditional Pap had a sensitivity of about 70% for detecting clinically significant precancerous lesions and cancer because of sampling and interpretive errors. The introduction of 2 liquid-based Pap smear collection systems in the 1990s, the use of HPV testing as a triage and co-testing with Pap smear, and the introduction of 2 automated screening devices have had a significant impact on improving the detection of such precancerous lesions. This review provides an analysis of the changes in Pap smear collection, improvements in screening, the evolutionary changes of high-risk HPV testing, reporting terminology of Pap smears, and clinical management guidelines. The future impact of 2 prophylactic HPV vaccines on the incidence of cervical carcinoma is also discussed. This article also discusses alternatives such as primary screening for high-risk HPV testing with visual inspection for cervical cancer detection used in resource-poor settings with a high incidence of cervical cancer.

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Response of Aldehyde Dehydrogenase (ALDH) after Cisplatin and All-Trans Retinoic Acid (ATRA) treatment of Spheroid-derived Cells (SDCs) from cervical cancer cell lines

10.1055/s-0038-1671355 ◽

2018 ◽

Cited By ~ 1

Author(s):

J Xu ◽

J Sehouli ◽

AM Kaufmann

Keyword(s):

Cervical Cancer ◽

Retinoic Acid ◽

Cell Lines ◽

Cancer Cell ◽

Aldehyde Dehydrogenase ◽

Cancer Cell Lines ◽

Cervical Cancer Cell ◽

Atra Treatment ◽

Trans Retinoic Acid ◽

All Trans Retinoic Acid

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Sensitization of cervical cancer cell lines to low-dose radiation by retinoic acid does not require functional p53

Gynecologic Oncology ◽

10.1016/j.ygyno.2004.12.034 ◽

2005 ◽

Vol 97 (1) ◽

pp. 142-150 ◽

Cited By ~ 3

Author(s):

Todd D. Tillmanns ◽

Scott A. Kamelle ◽

Suresh Guruswamy ◽

Natalie S. Gould ◽

Teresa L. Rutledge ◽

...

Keyword(s):

Cervical Cancer ◽

Retinoic Acid ◽

Cell Lines ◽

Cancer Cell ◽

Low Dose ◽

Cancer Cell Lines ◽

Cervical Cancer Cell ◽

Low Dose Radiation ◽

Dose Radiation

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Adenoviral Vectors Armed with PAPILLOMAVIRUs Oncogene Specific CRISPR/Cas9 Kill Human-Papillomavirus-Induced Cervical Cancer Cells

Cancers ◽

10.3390/cancers12071934 ◽

2020 ◽

Vol 12 (7) ◽

pp. 1934 ◽

Cited By ~ 1

Author(s):

Eric Ehrke-Schulz ◽

Sonja Heinemann ◽

Lukas Schulte ◽

Maren Schiwon ◽

Anja Ehrhardt

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

Adenoviral Vectors ◽

Human Papillomaviruses ◽

Cervical Cancer Cell ◽

Adenoviral Delivery ◽

Cervical Cancer Cells

Human papillomaviruses (HPV) cause malignant epithelial cancers including cervical carcinoma, non-melanoma skin and head and neck cancer. They drive tumor development through the expression of their oncoproteins E6 and E7. Designer nucleases were shown to be efficient to specifically destroy HPV16 and HPV18 oncogenes to induce cell cycle arrest and apoptosis. Here, we used high-capacity adenoviral vectors (HCAdVs) expressing the complete CRISPR/Cas9 machinery specific for HPV18-E6 or HPV16-E6. Cervical cancer cell lines SiHa and CaSki containing HPV16 and HeLa cells containing HPV18 genomes integrated into the cellular genome, as well as HPV-negative cancer cells were transduced with HPV-type-specific CRISPR-HCAdV. Upon adenoviral delivery, the expression of HPV-type-specific CRISPR/Cas9 resulted in decreased cell viability of HPV-positive cervical cancer cell lines, whereas HPV-negative cells were unaffected. Transduced cervical cancer cells showed increased apoptosis induction and decreased proliferation compared to untreated or HPV negative control cells. This suggests that HCAdV can serve as HPV-specific cancer gene therapeutic agents when armed with HPV-type-specific CRISPR/Cas9. Based on the versatility of the CRISPR/Cas9 system, we anticipate that our approach can contribute to personalized treatment options specific for the respective HPV type present in each individual tumor.

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HPV and Other Microbiota; Who’s Good and Who’s Bad: Effects of the Microbial Environment on the Development of Cervical Cancer—A Non-Systematic Review

Cells ◽

10.3390/cells10030714 ◽

2021 ◽

Vol 10 (3) ◽

pp. 714

Author(s):

Matthias Läsche ◽

Horst Urban ◽

Julia Gallwas ◽

Carsten Gründker

Keyword(s):

Systematic Review ◽

Cervical Cancer ◽

High Risk ◽

Cervical Carcinoma ◽

Hpv Infection ◽

Human Papillomaviruses ◽

Inflammatory Reactions ◽

Metabolic Signalling ◽

Microbial Environment ◽

High Risk Hpv

Cervical cancer is responsible for around 5% of all human cancers worldwide. It develops almost exclusively from an unsolved, persistent infection of the squamocolumnar transformation zone between the endo- and ecto-cervix with various high-risk (HR) human papillomaviruses (HPVs). The decisive turning point on the way to persistent HPV infection and malignant transformation is an immune system weakened by pathobionts and oxidative stress and an injury to the cervical mucosa, often caused by sexual activities. Through these injury and healing processes, HPV viruses, hijacking activated keratinocytes, move into the basal layers of the cervical epithelium and then continue their development towards the distal prickle cell layer (Stratum spinosum). The microbial microenvironment of the cervical tissue determines the tissue homeostasis and the integrity of the protective mucous layer through the maintenance of a healthy immune and metabolic signalling. Pathological microorganisms and the resulting dysbiosis disturb this signalling. Thus, pathological inflammatory reactions occur, which manifest the HPV infection. About 90% of all women contract an HPV infection in the course of their lives. In about 10% of cases, the virus persists and cervical intra-epithelial neoplasia (CIN) develops. Approximately 1% of women with a high-risk HPV infection incur a cervical carcinoma after 10 to 20 years. In this non-systematic review article, we summarise how the sexually and microbial mediated pathogenesis of the cervix proceeds through aberrant immune and metabolism signalling via CIN to cervical carcinoma. We show how both the virus and the cancer benefit from the same changes in the immune and metabolic environment.

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