Detection of HPV infection and molocular cytogenetic relationship between HPV DNA and c-myc gene in uterine cervical cancer cells.

ABT-737, a B cell lymphoma-2 (Bcl-2) family inhibitor, activates apoptosis in cancer cells. Arsenic trioxide is an apoptosis activator that impairs cancer cell survival. The aim of this study was to evaluate the effect of a combination treatment with ABT-737 and arsenic trioxide on uterine cervical cancer cells. MTT (3-(4,5-dimethylthiazol-2-yl)-25-diphenyltetrazolium bromide) assay revealed that ABT-737 and arsenic trioxide induced a synergistic effect on uterine cervical cancer cells. Arsenic trioxide enhanced ABT-737-induced apoptosis and caspase-7 activation and the ABT-737-mediated reduction of anti-apoptotic protein Mcl-1 in Caski cells. Western blot assay revealed that arsenic trioxide promoted the ABT-737-mediated reduction of CDK6 and thymidylate synthetase in Caski cells. Arsenic trioxide promoted ABT-737-inhibited mitochondrial membrane potential and ABT-737-inhibited ANT expression in Caski cells. However, ABT-737-elicited reactive oxygen species were not enhanced by arsenic trioxide. The combined treatment induced an anti-apoptosis autophagy in SiHa cells. This study is the first to demonstrate that a combination treatment with ABT-737 and arsenic trioxide induces a synergistic effect on uterine cervical cancer cells through apoptosis. Our findings provide new insights into uterine cervical cancer treatment.

Download Full-text

Optimisation of Folate-Mediated Liposomal Encapsulated Arsenic Trioxide for Treating HPV-Positive Cervical Cancer Cells In Vitro

International Journal of Molecular Sciences ◽

10.3390/ijms20092156 ◽

2019 ◽

Vol 20 (9) ◽

pp. 2156 ◽

Cited By ~ 5

Author(s):

Akhtar ◽

Ghali ◽

Wang ◽

Bell ◽

Li ◽

...

Keyword(s):

Cervical Cancer ◽

Folic Acid ◽

Cancer Cells ◽

Arsenic Trioxide ◽

Folate Receptor ◽

Hpv Infection ◽

Cytotoxicity Studies ◽

Cervical Cancer Cells ◽

Infected Cells

High-risk human papilloma virus (HPV) infection is directly associated with cervical cancer development. Arsenic trioxide (ATO), despite inducing apoptosis in HPV-infected cervical cancer cells in vitro, has been compromised by toxicity and poor pharmacokinetics in clinical trials. Therefore, to improve ATO’s therapeutic profile for HPV-related cancers, this study aims to explore the effects of length of ligand spacers of folate-targeted liposomes on the efficiency of ATO delivery to HPV-infected cells. Fluorescent ATO encapsulated liposomes with folic acid (FA) conjugated to two different PEG lengths (2000 Da and 5000 Da) were synthesised, and their cellular uptake was examined for HPV-positive HeLa and KB and HPV-negative HT-3 cells using confocal microscopy, flow cytometry, and spectrophotometer readings. Cellular arsenic quantification and anti-tumour efficacy was evaluated through inductively coupled plasma-mass spectrometry (ICP-MS) and cytotoxicity studies, respectively. Results showed that liposomes with a longer folic acid-polyethylene glycol (FA-PEG) spacer (5000 Da) displayed a higher efficiency in targeting folate receptor (FR) + HPV-infected cells without increasing any inherent cytotoxicity. Targeted liposomally delivered ATO also displayed superior selectivity and efficiency in inducing higher cell apoptosis in HPV-positive cells per unit of arsenic taken up than free ATO, in contrast to HT-3. These findings may hold promise in improving the management of HPV-associated cancers.

Download Full-text

The degradation of Rap1GAP via E6AP-mediated ubiquitin-proteasome pathway is associated with HPV16/18-infection in cervical cancer cells

Infectious Agents and Cancer ◽

10.1186/s13027-021-00409-9 ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Yinghui Wang ◽

Yihang Xie ◽

Boxuan Sun ◽

Yuwei Guo ◽

Ling Song ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Suppressor Gene ◽

Hpv Infection ◽

Degradation Pathway ◽

Human Papillomaviruses ◽

Siha Cells ◽

Cervical Cancer Cells ◽

Ubiquitin Proteasome ◽

Proteasome Pathway

Abstract Background Cervical cancers are closely associated with persistent high-risk human papillomaviruses (HR HPV) infection. The main mechanism involves the targeting of tumor suppressors, such as p53 and pRB, for degradation by HR HPV-encoded oncoproteins, thereby leading to tumorigenesis. Rap1GAP, a tumor suppressor gene, is down-regulated in many cancers. Previous studies have revealed that down-regulation of Rap1GAP is correlated with HPV16/18 infection in cervical cancer. However, the molecular mechanism remains unclear. In this study, we aimed to address the degradation pathway of Rap1GAP in HPV-positive cervical cancer cells. Methods HPV-positive (HeLa and SiHa) and negative (C33A) cervical cancer cells were used to analyze the pathways of Rap1GAP degradation. MG132 (carbobenzoxy-leucyl-leucyl-leucine) was used to inhibit protein degradation by proteasome. Co-immunoprecipitation (co-IP) was used to detect the interaction between Rap1GAP and E6AP. siRNA for E6AP was used to silence the expression of E6AP. Rapamycin was used to induce cell autophagy. Western blotting was used to check the levels of proteins. Results Following treatment with MG132, the levels of Rap1GAP were increased in the HR HPV-positive HeLa and SiHa cells, but not in the HPV-negative C33A cells. Co-immunoprecipitation assay revealed ubiquitinated Rap1GAP protein in HeLa and SiHa cells, but not in C33A cells. E6-associated protein (E6AP) mediated the ubiquitination of Rap1GAP by binding to it in HeLa and SiHa cells, but not in C33A cells. However, the levels of Rap1GAP were decreased in HeLa and SiHa cells after knocking down E6AP by siRNA. Silencing of E6AP did not affect the levels of Rap1GAP in C33A cells. Autophagy marker p62 was decreased and LC3 II/LC3 I was increased after knocking down E6AP in HeLa cells, but not in C33A cells. The levels of Rap1GAP were decreased after treating the cells with rapamycin to induce cell autophagy in HeLa and C33A cells. Conclusion Rap1GAP may be degraded by autophagy in cervical cancer cells, but HPV infection can switch the degradation pathway from autophagy to E6AP-mediated ubiquitin-proteasome degradation. E6AP may be a key component of the switch.

Download Full-text

Effects of ABT‑737 combined with irradiation treatment on uterine cervical cancer cells

Oncology Letters ◽

10.3892/ol.2019.10755 ◽

2019 ◽

Cited By ~ 1

Author(s):

Huang‑Pin Shen ◽

Wen‑Jun Wu ◽

Jiunn‑Liang Ko ◽

Tzu‑Fan Wu ◽

Shun‑Fa Yang ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Uterine Cervical Cancer ◽

Irradiation Treatment ◽

Cervical Cancer Cells

Download Full-text

Adenosine uptake is the major effector of extracellular ATP toxicity in human cervical cancer cells

Molecular Biology of the Cell ◽

10.1091/mbc.e14-01-0042 ◽

2014 ◽

Vol 25 (19) ◽

pp. 2905-2918 ◽

Cited By ~ 24

Author(s):

Paola de Andrade Mello ◽

Eduardo Cremonese Filippi-Chiela ◽

Jéssica Nascimento ◽

Aline Beckenkamp ◽

Danielle Bertodo Santana ◽

...

Keyword(s):

Cervical Cancer ◽

Cell Growth ◽

Cancer Cells ◽

Purinergic Signaling ◽

Hpv Infection ◽

Extracellular Atp ◽

Parp Cleavage ◽

Cervical Cancer Cells ◽

Human Cervical Cancer Cells ◽

Human Cervical Cancer

In cervical cancer, HPV infection and disruption of mechanisms involving cell growth, differentiation, and apoptosis are strictly linked with tumor progression and invasion. Tumor microenvironment is ATP and adenosine rich, suggesting a role for purinergic signaling in cancer cell growth and death. Here we investigate the effect of extracellular ATP on human cervical cancer cells. We find that extracellular ATP itself has a small cytotoxic effect, whereas adenosine formed from ATP degradation by ectonucleotidases is the main factor responsible for apoptosis induction. The level of P2×7 receptor seemed to define the main cytotoxic mechanism triggered by ATP, since ATP itself eliminated a small subpopulation of cells that express high P2×7 levels, probably through its activation. Corroborating these data, blockage or knockdown of P2×7 only slightly reduced ATP cytotoxicity. On the other hand, cell viability was almost totally recovered with dipyridamole, an adenosine transporter inhibitor. Moreover, ATP-induced apoptosis and signaling—p53 increase, AMPK activation, and PARP cleavage—as well as autophagy induction were also inhibited by dipyridamole. In addition, inhibition of adenosine conversion into AMP also blocked cell death, indicating that metabolization of intracellular adenosine originating from extracellular ATP is responsible for the main effects of the latter in human cervical cancer cells.

Download Full-text

Disruption of HPV16-E7 by CRISPR/Cas System Induces Apoptosis and Growth Inhibition in HPV16 Positive Human Cervical Cancer Cells

BioMed Research International ◽

10.1155/2014/612823 ◽

2014 ◽

Vol 2014 ◽

pp. 1-9 ◽

Cited By ~ 57

Author(s):

Zheng Hu ◽

Lan Yu ◽

Da Zhu ◽

Wencheng Ding ◽

Xiaoli Wang ◽

...

Keyword(s):

Cervical Cancer ◽

Growth Inhibition ◽

Cancer Cells ◽

Hpv Infection ◽

Hek293 Cells ◽

Guide Rna ◽

System A ◽

Cervical Cancer Cells ◽

E6 And E7 ◽

Human Cervical Cancer Cells

High-risk human papillomavirus (HR-HPV) has been recognized as a major causative agent for cervical cancer. Upon HPV infection, early genes E6 and E7 play important roles in maintaining malignant phenotype of cervical cancer cells. By using clustered regularly interspaced short palindromic repeats- (CRISPR-) associated protein system (CRISPR/Cas system), a widely used genome editing tool in many organisms, to target HPV16-E7 DNA in HPV positive cell lines, we showed for the first time that the HPV16-E7 single-guide RNA (sgRNA) guided CRISPR/Cas system could disrupt HPV16-E7 DNA at specific sites, inducing apoptosis and growth inhibition in HPV positive SiHa and Caski cells, but not in HPV negative C33A and HEK293 cells. Moreover, disruption of E7 DNA directly leads to downregulation of E7 protein and upregulation of tumor suppressor protein pRb. Therefore, our results suggest that HPV16-E7 gRNA guided CRISPR/Cas system might be used as a therapeutic strategy for the treatment of cervical cancer.

Download Full-text

The Dynamics of HPV Infection and Cervical Cancer Cells

Bulletin of Mathematical Biology ◽

10.1007/s11538-015-0124-2 ◽

2015 ◽

Vol 78 (1) ◽

pp. 4-20 ◽

Cited By ~ 10

Author(s):

Tri Sri Noor Asih ◽

Suzanne Lenhart ◽

Steven Wise ◽

Lina Aryati ◽

F. Adi-Kusumo ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Hpv Infection ◽

Cervical Cancer Cells

Download Full-text

Abnormal level of paxillin in cervical cancer cells is involved in tumor progression and invasion

Acta Biochimica Polonica ◽

10.18388/abp.2020_5379 ◽

2021 ◽

Author(s):

Hongqing Gu ◽

Jing Wen

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

Uterine Cervical Cancer ◽

Cancer Growth ◽

Over Expression ◽

Apoptosis Inhibition ◽

Cervical Cancer Cells ◽

Elevated Expression ◽

Abnormal Level

Human papillomavirus (HPV) is the primary causative agent for the uterine cervical cancer. The expression of oncoproteins E6/E7 promotes apoptosis inhibition and increases the risk of cervical cancer progression. Some research reported that elevated expression of paxillin (PXN) stimulated cancer growth and invasion. However, the clinical significance of PXN in cervical cancer has not been well characterized so far. We found that PXN mRNA expression and protein level are significantly upregulated in cervical cancer cells compared to adjacent normal cells. Furthermore, the paxillin over-expression was correlated with potential of tumorigenesis and invasion. Cervical cancer cells with increased paxillin expression had an ability to form more tumor clones and were characterized by higher invasiveness as well. Therefore, our findings suggest that paxillin may act as an important prognostic factor for cervical cancer patients as it promotes tumor regeneration and invasion.

Download Full-text