scholarly journals CRISPR/Cas9 nanoeditor of double knockout large fragments of E6 and E7 oncogenes for reversing drugs resistance in cervical cancer

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Xianhuang Li ◽  
Mingming Guo ◽  
Bei Hou ◽  
Bin Zheng ◽  
Zhiyun Wang ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Tumor Cells ◽  
Chemotherapy Resistance ◽  
Drug Tolerance ◽  
Double Knockout ◽  
Knock Out ◽  
Chemotherapy Drugs ◽  
E6 And E7 ◽  
Target Gene Therapy ◽  
Growth Of Tumor

AbstractDrug resistance of tumor cells is always a headache problem in clinical treatment. In order to combat chemotherapy-resistance in cervical cancer and improve treatment effect, we design a CRISPR/Cas9 nanoeditor to knock out two key oncogenes E6 and E7 that lead to drug tolerance. Meanwhile, the deletion of these two oncogenes can effectively reactivate p53 and pRB signaling pathways that inhibit the growth of tumor cells. Our results demonstrated the nanoeditor could simultaneously delete two oncogenes, and the size of DNA fragments knocked out reaches an unprecedented 563 bp. After the preparation of cationic liposomes combined with chemotherapy drug docetaxel (DOC), this nanosystem can significantly inhibit the drug tolerance of cancer cells and improve the therapeutic effect of cervical cancer. Therefore, this study provides a promising strategy for the treatment of cervical cancer by combining chemotherapy and double-target gene therapy. This strategy can also be applied in other disease models to customize personalized anti-tumor strategies by simply changing chemotherapy drugs and targeted genes.

scholarly journals Human Papillomavirus Type 16 (HPV-16) Virus-Like Particle L1-Specific CD8+ Cytotoxic T Lymphocytes (CTLs) Are Equally Effective as E7-Specific CD8+ CTLs in Killing Autologous HPV-16-Positive Tumor Cells in Cervical Cancer Patients: Implications for L1 Dendritic Cell-Based Therapeutic Vaccines

2009 ◽  
Vol 83 (13) ◽  
pp. 6779-6789 ◽  
Author(s):  
Stefania Bellone ◽  
Karim El-Sahwi ◽  
Emiliano Cocco ◽  
Francesca Casagrande ◽  
Marilisa Cargnelutti ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
T Lymphocytes ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Copy Number ◽  
Autologous Tumor ◽  
Hpv 16 ◽  
Expression Levels ◽  
E6 And E7

ABSTRACT Papillomavirus-like particles (VLPs) based on L1 capsid protein represent a promising prophylactic vaccine against human papillomavirus (HPV) infections. However, cell-mediated immune responses against this antigen are believed to be of limited therapeutic value in established HPV-infected cervical lesions and, for this reason, have not been intensively investigated in cervical cancer patients. In this study we analyzed and quantified by real-time PCR (RT-PCR) the RNA expression levels of E6, E7, and L1 genes in flash-frozen HPV-16 cervical carcinomas. In addition, the kinetics of expression of E6, E7, and L1 in HPV-16-infected primary cell lines established as long-term cultures in vitro was also evaluated at RNA and protein levels. Finally, in order to evaluate the therapeutic potential of L1-specific CD4+ and CD8+ T lymphocytes responses in cervical cancer patients, L1 VLP-loaded dendritic cells (DCs) were used to stimulate peripheral blood lymphocytes from cervical cancer patients and such responses were compared to those elicited by the E7 oncoprotein. We show that 22 of 22 (100%) flash-frozen cervical biopsy samples collected from HPV-16-positive cervical cancer patients harbor L1, in addition to E6 and E7 RNA, as detected by RT-PCR. E7 RNA copy number (mean, 176.2) was significantly higher in HPV-16-positive cervical cancers compared to the E6 RNA copy number (mean, 47.3) and the L1 copy number (mean, 58.3) (P < 0.0001 and P < 0.001, respectively). However, no significant differences in expression levels between E6 and L1 were found. Kinetic studies of E6, E7, and L1 RNA and protein expression levels in primary tumors showed a sharp reduction in L1 expression after multiple in vitro passages compared to E6 and E7. Autologous DCs pulsed with HPV-16 VLPs or recombinant full-length E7 elicited strong type 1 L1- and E7-specific responses in CD4+ and CD8+ T cells from cervical cancer patients. Importantly, L1 VLP-specific CD8+ T lymphocytes expressed strong cytolytic activity against autologous tumor cells and were as effective as E7-specific cytotoxic T lymphocytes in lysing naturally HPV-16-infected autologous tumor cells. Taken together, these data demonstrate a consistent expression of L1 in primary cervical tumors and the possibility of inducing effective L1/tumor-specific CD4+ and CD8+ T-lymphocyte responses in patients harboring HPV-infected cervical cancer. These results may have important implications for the treatment of patients harboring established HPV-infected lesions with L1 VLPs or combined E7/L1 DC-based vaccinations.

scholarly journals Multiobjective approach in the treatment of cancer

2020 ◽  
Author(s):  
Soukaina Sabir ◽  
Nadia Raissi ◽  
Mustapha Serhani
Keyword(s):  
Optimal Control ◽  
Tumor Cells ◽  
Pareto Front ◽  
Multi Objective Optimization ◽  
Pareto Solutions ◽  
Effector Cells ◽  
Multi Objective ◽  
Chemotherapy Drugs ◽  
Multiobjective Approach ◽  
Growth Of Tumor

In this work we deal with a cancer problem involving the growth of tumor cells and their interaction with effector cells. The goal is to find an optimal control minimizing tumor cells density together with the amount of chemotherapy drugs and maximizing the density of effector cells. By invoking the multi-objective optimization we characterize optimal Pareto solutions and give simulation of Pareto front.

Effect on humoral and cellular immunity and on apoptosis in CIN2, CIN3, and HPV16+ cervical cancer of therapeutic divalent genetic vaccination with CMV replicon system (CRS) delivering HPV16 recombinantly mutated E6 and E7 viral oncogenes targeting p53 and Rb, respectively

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. 3062-3062
Author(s):  
J. Giannios ◽  
J. Peristeris ◽  
N. Alexandropoulos ◽  
T. Kononas ◽  
P. Ginopoulos
Keyword(s):  
Cervical Cancer ◽  
Immune Responses ◽  
Cancer Cells ◽  
Tumor Cells ◽  
Binding Sites ◽  
Injection System ◽  
Cellular Immune Responses ◽  
Cervical Cancer Cells ◽  
E6 And E7 ◽  
Cellular Immune

3062 Background: Prophylactic vaccines have no therapeutic capacity for all the women who are already infected with HPV16 and have developed cervical intraepithelial neoplasia (CIN) or cervical cancer. Approximately 300 million women with CIN and cervical cancer will require therapy in the next decades. Thus, there is a great demand for a therapeutic HPV vaccine. Methods: We developed a cytomegalovirus (CMV) replicon system (CRS) for delivery of the HPV16 recombinantly mutated E6 and E7 genes replacing part of the CMV genome for the HPV genes, which were genetically altered to block binding sites for p53 and Rb.The replicon-vectors infected and co-transfected CIN and cervical cancer cells in animal models derived from HPV16(+) CIN, and cervical Ca cells obtained from patients. The genetic vaccine was administered subcutaneously (SC) with a needless injection system. Results: After vaccination,the viral E6 oncogene did not degrade apoptotic p53, and it blocked activation of telomerase. This induced apoptosis and DNA repair in CIN and cervical cancer cells. Furthermore, the E7 viral oncogene did not degrade the retinoblastoma oncogene (Rb) protein releasing transcription factor E2F. This vaccination led to scheduled cell cycle entry, genetic stability, and mortalization of tumor cells. Humoral and cellular immune responses were exhibited, which led to irreversible D2 apoptotic stage of PCD type I leading to a bystander killing effect of CIN, and cervical cancer cells. BrdU and MTT analysis exhibited inhibition of DNA synthesis and metabolic activity of vaccinated tumor cells compared to controls. Conclusions: This genetic divalent vaccine coding for E6 and E7 mutations designed to prevent p53 and Rb binding sites activated humoral and cellular immune responses leading to apoptosis of CIN and cervical cancer cells. It is up to translational medicine to bring this therapeutic vaccine into the clinic for patients with CIN2, CIN3, and cervical cancer patients. No significant financial relationships to disclose.

E6 and E7 oncoproteins: Potential targets of cervical cancer

2020 ◽  
Vol 27 ◽  
Author(s):  
Ramarao Malla ◽  
Mohammad Amjad Kamal
Keyword(s):  
Cervical Cancer ◽  
Drug Resistance ◽  
Host Cell ◽  
Molecular Mechanisms ◽  
Immune Therapy ◽  
Cervical Lesions ◽  
Diagnostic Strategies ◽  
E6 And E7 Oncoproteins ◽  
E6 And E7

: Cervical cancer (CC) is the fourth leading cancer in women in the age group 15-44 globally. Experimental as well as epidemiological studies identified that type16 and 18 HPV cause 70% of precancerous cervical lesions as well as cervical cancer worldwide by bringing about genetic as well as epigenetic changes in the host genome. The insertion of the HPV genome triggers various defense mechanisms including the silencing of tumor suppressor genes as well as activation of oncogenes associated with cancer metastatic pathway. E6 and E7 are small oncoproteins consisting of 150 and 100 amino acids respectively. These oncoproteins affect the regulation of the host cell cycle by interfering with p53 and pRb. Further these oncoproteins adversely affect the normal functions of the host cell by binding to their signaling proteins. Recent studies demonstrated that E6 and E7 oncoproteins are potential targets for CC. Therefore, this review discusses the role of E6 and E7 oncoproteins in metastasis and drug resistance as well as their regulation, early oncogene mediated signaling pathways. This review also uncovers the recent updates on molecular mechanisms of E6 and E7 mediated phytotherapy, gene therapy, immune therapy, and vaccine strategies as well as diagnosis through precision testing. Therefore, understanding the potential role of E6/E7 in metastasis and drug resistance along with targeted treatment, vaccine, and precision diagnostic strategies could be useful for the prevention and treatment of cervical cancer.

scholarly journals The Role of Glycolysis in the Growth of Tumor Cells

1961 ◽  
Vol 236 (2) ◽  
pp. 285-288
Author(s):  
John Papaconstantinou ◽  
Sidney P. Colowick
Keyword(s):  
Tumor Cells ◽  
Growth Of Tumor

scholarly journals The Role of Glycolysis in the Growth of Tumor Cells

1965 ◽  
Vol 240 (7) ◽  
pp. 2791-2796
Author(s):  
Edward B. Goldberg ◽  
Harold M. Nitowsky ◽  
Sidney P. Colowick
Keyword(s):  
Tumor Cells ◽  
Growth Of Tumor

scholarly journals Association of cervical carcinogenesis risk with HPV16 E6 and E7 variants in the Taizhou area, China

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Mei-Zhen Dai ◽  
Yi Qiu ◽  
Xing-Hong Di ◽  
Wei-Wu Shi ◽  
Hui-Hui Xu
Keyword(s):  
Cervical Cancer ◽  
Cervical Carcinogenesis ◽  
Chi Square ◽  
Major Health Problem ◽  
Hpv16 E6 ◽  
Exact Test ◽  
Chi Square Test ◽  
E6 And E7

Abstract Background Human papillomavirus (HPV) type 16 accounts for a larger share of cervical cancer and has been a major health problem worldwide for decades. The progression of initial infection to cervical cancer has been linked to viral sequence properties; however, the role of HPV16 variants in the risk of cervical carcinogenesis, especially with longitudinal follow-up, is not fully understood in China. Methods We aimed to investigate the genetic variability of HPV16 E6 and E7 oncogenes in isolates from cervical exfoliated cells. Between December 2012 and December 2014, a total of 310 single HPV16-positive samples were selected from women living in the Taizhou area, China. Sequences of all E6 and E7 oncogenes were analysed by PCR-sequencing assay. Detailed sequence comparison, genetic heterogeneity analyses and maximum-likelihood phylogenetic tree construction were performed with BioEdit Sequence Alignment Editor and MEGA X software. Data for cytology tests and histological diagnoses were obtained from our Taizhou Area Study with longitudinal follow-up for at least 5 years. The relationship between HPV16 variants and cervical carcinogenesis risk was analysed by the chi-square test or Fisher’s exact test. Results In this study, we obtained 64 distinct variation patterns with the accession GenBank numbers MT681266-MT681329. Phylogenetic analysis revealed that 98.3% of HPV16 variants belong to lineage A, in which the A4 (Asian) sublineage was dominant (64.8%), followed by A2 (12.1%), A1 (11.4%), and A3 (10.0%). The A4 (Asian) sublineage had a higher risk of CIN2+ than the A1–3 (European) sublineages (OR = 2.69, 95% CI = 1.04–6.97, P < 0.05). Furthermore, nucleotide variation in HPV16 E6 T178G is associated with the development of cervical cancer. Conclusion These data could provide novel insights into the role of HPV16 variants in cervical carcinogenesis risk in China.

scholarly journals ATRT-14. MACROPHAGE-TUMOR CELL INTERACTION PROMOTES ATRT PROGRESSION AND CHEMORESISTANCE

2020 ◽  
Vol 22 (Supplement_3) ◽  
pp. iii278-iii278
Author(s):  
Viktoria Melcher ◽  
Monika Graf ◽  
Marta Interlandi ◽  
Natalia Moreno ◽  
Flavia W de Faria ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Tumor Cells ◽  
Chemotherapy Resistance ◽  
Cell Communication ◽  
Xenograft Model ◽  
Cell Interaction ◽  
Immunofluorescent Staining ◽  
Genetic Traits ◽  
Rhabdoid Tumors

Abstract Atypical teratoid/rhabdoid tumors (ATRT) are pediatric brain neoplasms that are known for their heterogeneity concerning pathophysiology and outcome. The three genetically rather uniform but epigenetically distinct molecular subgroups of ATRT alone do not sufficiently explain the clinical heterogeneity. Therefore, we examined the tumor microenvironment (TME) in the context of tumor diversity. By using multiplex-immunofluorescent staining and single-cell RNA sequencing (scRNA-seq) we unveiled the pan-macrophage marker CD68 as a subgroup-independent negative prognostic marker for survival of ATRT patients. ScRNA-seq analysis of murine ATRT-SHH, ATRT-MYC and extracranial RT (eRT) provide a delineation of the TME, which is predominantly infiltrated by myeloid cells: more specifically a microglia-enriched niche in ATRT-SHH and a bone marrow-derived macrophage infiltration in ATRT-MYC and eRT. Exploring the cell-cell communication of tumor cells with tumor-associated immune cells, we found that Cd68+ tumor-associated macrophages (TAMs) are central to intercellular communication with tumor cells. Moreover, we uncovered distinct tumor phenotypes in murine ATRT-MYC that share genetic traits with TAMs. These intermediary cells considerably increase the intratumoral heterogeneity of ATRT-MYC tumors. In vitro co-culture experiments recapitulated the capability of ATRT-MYC cells to interchange cell material with macrophages extensively, in contrast to ATRT-SHH cells. We found that microglia are less involved in the exchange of information with ATRT cells and that direct contact is a prerequisite for incorporation. A relapse xenograft model implied that intermediary cells are involved in the acquisition of chemotherapy resistance. We show evidence that TAM-tumor cell interaction is one mechanism of chemotherapy resistance and relapse in ATRT.

Effect of Garlic, Chinese Medicinal Drugs and Amino Acids on Growth of Erlich Ascites Tumor Cells in Mice

1983 ◽  
Vol 11 (01n04) ◽  
pp. 69-73 ◽  
Author(s):  
Y.M. Choy ◽  
T.T. Kwok ◽  
K.P. Fung ◽  
C.Y. Lee
Keyword(s):  
Amino Acids ◽  
Tumor Cells ◽  
Peritoneal Cavity ◽  
Growth And Development ◽  
Ehrlich Ascites Tumor Cells ◽  
Ascites Tumor ◽  
Tumor Bearing ◽  
Ehrlich Ascites ◽  
Medicinal Drugs ◽  
Growth Of Tumor

A number of food materials or drugs have been screened for the effect on the growth and development of transplantable Ehrlich ascites tumor cells. Growth of tumor-bearing mice was significantly inhibited by feeding garlic as well as some amino acids. These materials significantly reduced the total number of free tumor cells growing in the peritoneal cavity of mice and prolonged significantly the length of time for 50% death of tumor-bearing mice.

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