Cytotoxic Effect of Disulfiram/Copper On Human Cervical Cancer Cell Lines and LGR5-Positive Cancer Stem-Like Cells

Abstract Background: Tumor resistance is a global challenge for tumor treatment. Cancer stem cells (CSCs) are the main population of tumor cells for drug resistance. We have reported that high aldehyde dehydrogenase (ALDH) activity represents a functional marker for cervical CSCs. Here we aim at disulfiram (DS), an ALDH inhibitor, that has the potential as a novel treatment to be used for cervical cancer.Methods: MTT assay, western blot, FCS analysis and sorting, vector construction and transfection, in vivo anti-tumor assays were performed using cervical cancer cell lines SiHa and HeLa. Cell cycle distribution and cell apoptosis were carried out by flow cytometry. The cytotoxicity of DS was detected by MTT assay and a xenograft cervical cancer model. Results: Disulfiram was cytotoxic to cervical cancer cell lines in a copper (Cu)-dependent manner. Disulfiram/copper (DS/Cu) complex induced deregulation of S-phase and inhibited the expression of stemness marker in cervical cancer cells. DS/Cu caused the death of LGR5-positive cervical cancer cells, a cancer stem-like cell population, which lead to cisplatin resistance in cervical cancer cells. Furthermore, DS/Cu complex had the greater antitumor efficacy on cervical cancer than cisplatin group in vitro and in vivo. Conclusion: Our findings indicate that the cytotoxicity of DS/Cu complex may be superior to cisplatin because of targeting LGR5-positive cervical cancer stem-like cells in cervical cancer. Thus, the DS/Cu complex may represent a potential therapeutic strategy for cervical cancer patients.

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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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LncRNA SNHG1 enhances cell proliferation, migration, and invasion in cervical cancer

Biochemistry and Cell Biology ◽

10.1139/bcb-2017-0188 ◽

2018 ◽

Vol 96 (1) ◽

pp. 38-43 ◽

Cited By ~ 34

Author(s):

Yang Liu ◽

Yanling Yang ◽

Lei Li ◽

Yuan Liu ◽

Peng Geng ◽

...

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Cell Migration ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

Cervical Cancer Cell ◽

Knock Down ◽

Cervical Cancer Cells

Objective: This study investigated the effects of lncRNA SNHG1 on the proliferation, migration, and invasiveness of cervical cancer cells. Methods: Three pairs of cervical cancer tissue samples and their corresponding adjacent samples were analyzed using Human LncRNA Microarray V3.0 chip for differential analysis. The expression of SNHG1 in cervical cancer cell lines was verified by qRT–PCR. CCK8 assays and colony formation assays were used to study the changes in cell proliferation. Cell migration and Transwell assays were used to study changes in cell migration and invasiveness. Results: SNHG1 was highly expressed in cervical cancer tissues and cervical cancer cell lines. SNHG1 siRNA could knock-down the expression level of SNHG1 in cervical cancer cell lines HeLa and C33-A. After knock-down of SNHG1, cell proliferation and migration as well as invasiveness in HeLa and C-33A cells decreased. Conclusion: LncRNA SNHG1 promotes the development of cervical cancer cells.

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Doxycycline inhibits proliferation and induces apoptosis of both human papillomavirus positive and negative cervical cancer cell lines

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2015-0481 ◽

2016 ◽

Vol 94 (5) ◽

pp. 526-533 ◽

Cited By ~ 13

Author(s):

Yan Zhao ◽

Xinyu Wang ◽

Lei Li ◽

Changzhong Li

Keyword(s):

Cervical Cancer ◽

Human Papillomavirus ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

Cervical Cancer Cell ◽

Cervical Cancer Cells

The clinical management of cervical cancer remains a challenge and the development of new treatment strategies merits attention. However, the discovery and development of novel compounds can be a long and labourious process. Drug repositioning may circumvent this process and facilitate the rapid translation of hypothesis-driven science into the clinics. In this work, we show that a FDA-approved antibiotic, doxycycline, effectively targets human papillomavirus (HPV) positive and negative cervical cancer cells in vitro and in vivo. Doxycycline significantly inhibits proliferation of a panel of cervical cancer cell lines. It also induces apoptosis of cervical cancer cells in a time- and dose-dependent manner. In addition, the apoptosis induced by doxycycline is through caspase-dependent pathway. Mechanism studies demonstrate that doxycycline affects oxygen consumption rate, glycolysis, and reduces ATP levels in cervical cancer cells. In HeLa xenograft mouse model, doxycycline significantly inhibits growth of tumour. Our in vitro and in vivo data clearly demonstrate the inhibitory effects of doxycycline on the growth and survival of cervical cancer cells. Our work provides the evidence that doxycycline can be repurposed for the treatment of cervical cancer and targeting energy metabolism may represent a potential therapeutic strategy for cervical cancer.

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Proteomics Analysis of Andrographolide-Induced Apoptosis Via the Regulation of Tumor Suppressor p53 Proteolysis in Cervical Cancer-Derived Human Papillomavirus 16-Positive Cell Lines

International Journal of Molecular Sciences ◽

10.3390/ijms22136806 ◽

2021 ◽

Vol 22 (13) ◽

pp. 6806

Author(s):

Pariyakorn Udomwan ◽

Chamsai Pientong ◽

Panwad Tongchai ◽

Ati Burassakarn ◽

Nuchsupha Sunthamala ◽

...

Keyword(s):

Cervical Cancer ◽

Tumor Suppressor ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

Human Papillomaviruses ◽

Cervical Cancer Cell ◽

World Population ◽

Cervical Cancer Cells

Regardless of the prophylactic vaccine accessibility, persistent infections of high-risk human papillomaviruses (hr-HPVs), recognized as an etiology of cervical cancers, continues to represent a major health problem for the world population. An overexpression of viral early protein 6 (E6) is linked to carcinogenesis. E6 induces anti-apoptosis by degrading tumor suppressor proteins p53 (p53) via E6-E6-associated protein (E6AP)-mediated polyubiquitination. Thus, the restoration of apoptosis by interfering with the E6 function has been proposed as a selective medicinal strategy. This study aimed to determine the activities of andrographolide (Androg) on the disturbance of E6-mediated p53 degradation in cervical cancer cell lines using a proteomic approach. These results demonstrated that Androg could restore the intracellular p53 level, leading to apoptosis-induced cell death in HPV16-positive cervical cancer cell lines, SiHa and CaSki. Mechanistically, the anti-tumor activity of Androg essentially relied on the reduction in host cell proteins, which are associated with ubiquitin-mediated proteolysis pathways, particularly HERC4 and SMURF2. They are gradually suppressed in Androg-treated HPV16-positive cervical cancer cells. Collectively, the restoration of p53 in HPV16-positive cervical cancer cells might be achieved by disruption of E3 ubiquitin ligase activity by Androg, which could be an alternative treatment for HPV-associated epithelial lesions.

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Anti-Silencing Function 1B Overexpression Affects Prognosis and Promotes Invasion in Cervical Carcinoma via Activation of Wnt/β-Catenin Signaling Pathway

10.21203/rs.3.rs-345176/v1 ◽

2021 ◽

Author(s):

Weijing Zhang ◽

Han Li ◽

Xiaoying Sun ◽

Yongjie Shi ◽

Yadi Yang ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Western Blotting ◽

Cervical Carcinoma ◽

Cancer Cell Lines ◽

Cervical Cancer Cell ◽

Clinicopathological Parameters ◽

Cervical Cancer Cells

Abstract Background: Anti-silencing function 1B histone chaperone (ASF1B) has been identified to compensate for growth defects and sensitivity to replication stress. Recent studies demonstrated that ASF1B plays an important role in the tumorigenesis of human cancer. However, its relationship with clinical outcome of cervical carcinoma is unknown. Therefore, we aimed to investigate the expression pattern and clinical significance of ASF1B in cervical cancer and its role in regulating invasion and metastasis of cervical cancer cell lines.Methods: Expression of ASF1B was analyzed in eight cervical cancer cell lines, and eight pairs of cervical cancer samples and the adjacent normal specimens, using real-time PCR and western blotting. Immunohistochemistry was used to analyze ASF1B expression in paraffin-embedded tissues from 147 cervical cancer patients. The association between ASF1B expression levels, clinicopathological parameters, and prognosis was analyzed statistically. Moreover, the biological function and potential mechanism of ASF1B in migration and invasion of cervical cancer cells were investigated by in vitro experiments and Western blotting.Results: ASF1B mRNA and protein levels were overexpressed in cervical cancer cell lines and tissues compared with normal cells and adjacent normal cervical specimens. In paraffin-embedded cervical carcinoma tissues, upregulation of ASF1B protein was identified in 86 (58.5%) of 147 cases, and was remarkably associated with pelvic lymph node metastasis, lymphovascular space involvement, property of surgical margin, FIGO stage, sqaumous cell carcinoma antigen and poor survival. Cox analysis demonstrated that ASF1B expression was an independent risk predictor for survival in patients with cervical carcinoma. Additionally, down-regulation of ASF1B significantly inhibited invasion and migration of cervical cancer cells. Moreover, it was demonstrated that ASF1B regulated the Wnt/β-catenin pathway in cervical carcinoma.Conclusions: This study suggested that ASF1B might serve as a important prognostic biomarker and therapeutic target for patients with cervical carcinoma.

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Four electrode-based impedimetric biosensors for evaluating cytotoxicity of tamoxifen on cervical cancer cells

RSC Advances ◽

10.1039/d0ra09155c ◽

2021 ◽

Vol 11 (2) ◽

pp. 798-806

Author(s):

Rangadhar Pradhan ◽

Ashish Kalkal ◽

Shlok Jindal ◽

Gopinath Packirisamy ◽

Sanjeev Manhas

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Cancer Cell Lines ◽

Cervical Cancer Cell ◽

Cervical Cancer Cells

In the current study, novel four electrode-based impedimetric biosensors have been fabricated using photolithography techniques and utilized to evaluate the cytotoxicity of tamoxifen on cervical cancer cell lines.

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Human papillomavirus type 18 oncoproteins exert their oncogenicity in esophageal and tongue squamous cell carcinoma cell lines distinctly

10.21203/rs.2.14671/v1 ◽

2019 ◽

Author(s):

Siaw Shi Boon ◽

Zigui Chen ◽

Karen Lee Yin Ching ◽

Liuyang Cai ◽

Jintao Li ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Tongue Cancer ◽

Cancer Cell Lines ◽

Cervical Cancer Cells ◽

Esophageal Cancer Cell ◽

Hpv18 E6 ◽

E6 And E7

Abstract Background: Increasing evidence indicates an etiological role of human papillomavirus (HPV) in head and neck cancers, particularly oropharyngeal squamous cell carcinoma (OPSCC). However, the association between HPV and other cancers, including esophageal and tongue remains unclear. Methods: We compared the molecular characteristics of HPV18 E6 and E7 in esophageal (EC109 and EC9706) and tongue (Tca83) cancer cell lines with reference to cervical cancer (HeLa). We studied the expression of HPV transcripts using Next Generation RNA sequencing. We used small interference RNA (siRNA) against HPV18 oncoproteins to verify their oncogenic roles in molecular signaling, targeting metalloproteases and apoptotic pathways in these cancers. Results: We observed that the HPV transcription profiles of esophageal and tongue cancer cells mimicked that of cervical cancer cells, with notable disruption of E2, and expression of E6, spliced E6 (E6 * ), E7, E1 and L1 transcripts. As with cervical cancer cells, p53 and its downstream transactivation target, p21, were found to be the major targets of E6 in esophageal and tongue cancer cell lines. Intriguingly, E7 preferentially targeted p130 in the two esophageal cancer cell lines, instead of pRb as in cervical cancer. Tca83 exhibited an E7 to E6 transcript ratio comparable to HeLa (cervix), targeted the ERK1/2 and MMP2 pathways, and was dependent on E6 and E7 to survive and proliferate. In contrast, both the esophageal cancer cell lines were distinct from HeLa in these aspects. Conclusion: This is the first study to delineate the transcripts expression and role of HPV18 E6 and E7 in esophageal and tongue cancer cell lines. This findings suggest that HPV18 plays an oncogenic role in inducing these cancers, albeit via distinct pathways than those observed in cervical cancer.

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Ectonucleotidase expression profile and activity in human cervical cancer cell lines

Biochemistry and Cell Biology ◽

10.1139/bcb-2013-0051 ◽

2014 ◽

Vol 92 (2) ◽

pp. 95-104 ◽

Cited By ~ 9

Author(s):

Aline Beckenkamp ◽

Danielle Bertodo Santana ◽

Alessandra Nejar Bruno ◽

Luciane Noal Calil ◽

Emerson André Casali ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Adenine Nucleotide ◽

Expression Profiles ◽

Cancer Cell Lines ◽

Cervical Cancer Cell ◽

Human Cervical Cancer Cell ◽

Human Cervical Cancer

Cervical cancer is the third most frequent cancer in women worldwide. Adenine nucleotide signaling is modulated by the ectonucleotidases that act in sequence, forming an enzymatic cascade. Considering the relationship between the purinergic signaling and cancer, we studied the E-NTPDases, ecto-5′-nucleotidase, and E-NPPs in human cervical cancer cell lines and keratinocytes. We evaluated the expression profiles of these enzymes using RT-PCR and quantitative real-time PCR analysis. The activities of these enzymes were examined using ATP, ADP, AMP, and p-nitrophenyl-5′-thymidine monophosphate (p-Nph-5′-TMP) as substrate, in a colorimetric assay. The extracellular adenine nucleotide hydrolysis was estimated by HPLC analysis. The hydrolysis of all substrates exhibited a linear pattern and these activities were cation-dependent. An interesting difference in the degradation rate was observed between cervical cancer cell lines SiHa, HeLa, and C33A and normal imortalized keratinocytes, HaCaT cells. The mRNA of ecto-5′-nucleotidase, E-NTPDases 5 and 6 were detectable in all cell lines, and the dominant gene expressed was the Entpd 5 enzyme, in SiHa cell line (HPV16 positive). In accordance with this result, a higher hydrolysis activity for UDP and GDP nucleotides was observed in the supernatant of the SiHa cells. Both normal and cancer cells presented activity and mRNAs of members of the NPP family. Considering that these enzymes exert an important catalytic activity, controlling purinergic nucleotide concentrations in tumors, the presence of ectonucleotidases in cervical cancer cells can be important to regulate the levels of extracellular adenine nucleotides, limiting their effects.

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Histone Deacetylation Regulated by KDM1A to Suppress DACT1 in Proliferation and Migration of Cervical Cancer

Analytical Cellular Pathology ◽

10.1155/2021/5555452 ◽

2021 ◽

Vol 2021 ◽

pp. 1-16

Author(s):

Lingjuan Zeng ◽

Chunyan Chen ◽

Chanjiao Yao

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Histone Deacetylation ◽

Cervical Cancer Cell ◽

Migration Ability ◽

Cervical Cancer Cells ◽

And Migration ◽

Proliferation And Migration

Objective. Increased expression of KDM1A and decreased expression of DACT1 in cervical cancer cells were noticed in a previous study. This study is aimed at exploring the mechanism behind the KDM1A regulation on DACT1 in cervical cancer cells. Methods. The expression profile of KDM1A and DACT1 in cervical cancer tissues was searched in TCGA database. In vitro experiments verified the effect of KDM1A and DACT1 on proliferation and migration ability of cervical cancer cell lines after cell transfection. The interaction of KDM1A with HDAC1 was identified by coimmunoprecipitation (Co-IP). The expression levels of KDM1A and DACT1 in cervical cancer cell lines were determined by qRT-PCR and western blot. Results. TCGA database showed that cervical cancer tissues had elevated expression of KDM1A and decreased expression of DACT1, which was consistent with the observation in cervical cancer cell lines. KDM1A was found to negatively regulate DACT1 through histone deacetylation. Meanwhile, the downregulation of KDM1A or overexpression of DACT1 could suppress the cell proliferation and migration ability in HeLa and SiHa cells. Cotransfection of KDM1A and DACT1 overexpression could reverse the increased cell proliferation and migration ability induced by KDM1A overexpression. Conclusion. KDM1A can downregulate DACT1 expression through histone deacetylation and therefore suppress the proliferation and migration of cervical cancer cells.

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Human papillomavirus type 18 oncoproteins exert their oncogenicity in esophageal and tongue squamous cell carcinoma cell lines distinctly

10.21203/rs.2.14671/v2 ◽

2019 ◽

Author(s):

Siaw Shi Boon ◽

Zigui Chen ◽

Jintao Li ◽

Karen Lee Yin Ching ◽

Liuyang Cai ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Cancer Cell ◽

Tongue Cancer ◽

Cancer Cell Lines ◽

Cervical Cancer Cells ◽

Esophageal Cancer Cell ◽

Hpv18 E6 ◽

E6 And E7

Abstract Increasing evidence indicates an etiological role of human papillomavirus (HPV) in head and neck cancers, particularly oropharyngeal squamous cell carcinoma (OPSCC). However, the association between HPV and other cancers, including esophageal and tongue remains unclear. This study delineated the molecular characteristics of HPV18 E6 and E7 in esophageal (EC109 and EC9706) and tongue (Tca83) cancer cell lines with reference to cervical cancer (HeLa). Overall, the HPV transcription profiles of esophageal and tongue cancer cells mimicked that of cervical cancer cells, with notable disruption of E2, and expression of E6, spliced E6 (E6*), E7, E1 and L1 transcripts. As with cervical cancer cells, p53 and its downstream transactivation target, p21, were found to be the major targets of E6 in esophageal and tongue cancer cell lines. Intriguingly, E7 preferentially targeted p130 in the two esophageal cancer cell lines, instead of pRb as in cervical cancer. Tca83 exhibited an E7 to E6 transcript ratio comparable to HeLa (cervix), targeted the ERK1/2 and MMP2 pathways, and was dependent on E6 and E7 to survive and proliferate. In contrast, both the esophageal cancer cell lines were distinct from HeLa in these aspects. In conclusion, this is the first study that delineates transcript expression and protein interaction of HPV18 E6 and E7 in esophageal and tongue cancer cell lines, suggesting that HPV plays a role in inducing these cancers, albeit via distinct pathways than those observed in cervical cancer.

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