scholarly journals Expression of the TIMP2 gene is not regulated by promoter hypermethylation in the Caski cell line

2012 ◽  
Vol 3 (5) ◽  
pp. 1079-1082 ◽  
Author(s):  
GAURAV PARASHAR ◽  
NEENA CAPALASH

2020 ◽  
Vol 8 (1) ◽  
pp. 12 ◽  
Author(s):  
Sachin Bhoora ◽  
Yuvelia Pather ◽  
Sumari Marais ◽  
Rivak Punchoo

Vitamin D has displayed anti-cancer actions in numerous in vitro studies. Here, we investigated the anti-cancer actions of cholecalciferol, a vitamin D precursor, on a metastatic cervical cancer cell line, namely, CaSki. Experimental cultures were incubated for 72 h and treated with cholecalciferol (10–1000 ng/mL). In the present study, cell count, viability, proliferation and cell cycle were analyzed by a crystal violet assay, trypan blue assay, Ki67 proliferation, and a cell cycle assay, respectively. Biomarkers of apoptosis, necrosis, and autophagic cell death were measured by the Caspase 3/7 and Annexin V/7-AAD Muse™ assays, a LC3-II assay, and a lactate dehydrogenase release assay, respectively. The ultrastructural features of cell death were assessed by transmission electron microscopy. A statistical analysis was performed using a one-way ANOVA and Bonferroni’s post-hoc analysis test, and p < 0.05 is considered statistically significant here. The results identify statistical decreases in cell count and viability at high-dose treatments (100 and 1000 ng/mL). In addition, significant increases in apoptotic biochemical markers and apoptotic ultrastructure are shown to be present at high-dose treatments. In conclusion, high-dose cholecalciferol treatments inhibit cell count and viability, which are both mediated by apoptotic induction in the CaSki cell line.



2014 ◽  
Vol 18 (03) ◽  
pp. 182-187 ◽  
Author(s):  
Pankaj Kumar Chaturvedi ◽  
Yong-Wan Kim ◽  
Sang Soo Kim ◽  
Woong Shick Ahn

Photodynamic therapy (PDT) is a promising modality in both the curative and palliative treatment against a variety of experimental and naturally occurring human cancers. At present, chlorophyll a derivatives are extensively used for the synthesis of photosensitizers (PSs) for PDT of tumors. In the present study, chlorophyll-a was extracted from the blue-green algae Spirulina platensis by refluxing with acetone. The extract was further acid treated to obtain methylpheophorbide-a (MPa), which was then refluxed in collidine and methylpyropheophorbide-a (Mppa) was obtained. After that, Mppa was converted to pyropheophorbide-a (Ppa) by treatment with 50% sulfuric acid. Finally, phototoxicity and dark toxicity of purified Ppa in two different cell lines, TC-1 and CaSki, were examined by MTT assay. The results suggest that Ppa is more toxic to TC-1 cell line than CaSki cell line. In vivo, the photosensitizing efficiency of Ppa was also higher than those of unloaded PS. These results indicate the potential of Ppa in PDT.



1997 ◽  
Vol 65 (2) ◽  
pp. 287-298 ◽  
Author(s):  
Tzyy-Choou Wu ◽  
Sung-Tsang Hsieh ◽  
Benjamin W. Purow ◽  
Robert J. Kurman


Author(s):  
Dr. Reena Rani ◽  
Dr. Anupam Kumar Singh ◽  
Dr. Samreen Khan ◽  
Prof. Najmul Islam

Background & objectives: Resveratrol (trans-3,4,5’ –trihydroxystilbene) is an active compound in food, such as redgrapes, peanuts, and berries. A body of evidence shows that resveratrol is able to inhibit thegrowth of many cancers such as leukemia, breast cancer and primary brain tumors. The objective of the present study was to demonstrate the dose and time dependent apoptotic effect of Resveratrol on Human Caski cell line. Methods: We used cell viability assays like MTT and Trypan blue assays to demonstrate the inhibition of cancer cell proliferation in a dose and time dependent manner. Results: Our results have shown that Resveratrol inhibited proliferation of Caski cells in a concentration and time dependent manner in 48 hr cultures. No significant effect was observed in 24 hour cultures of cell viability assays. Interpretation & conclusions: The results indicated that Resveratrol and NAC inhibited cell growth in Caski cells as a function of dose and time. Keywords: Resveratrol , Cervical cancer, apoptosis



2012 ◽  
Vol 11 (4) ◽  
pp. 237-242 ◽  
Author(s):  
Zhiguo Rao ◽  
Jianfei Gao ◽  
Bicheng Zhang ◽  
Bo Yang ◽  
Jiren Zhang


2022 ◽  
Vol 12 ◽  
Author(s):  
Shuaibing Yang ◽  
Qianqian Zhao ◽  
Lihua Tang ◽  
Zejia Chen ◽  
Zhaoting Wu ◽  
...  

Human papillomavirus (HPV) is a causal agent for most cervical cancers. The physical status of the HPV genome in these cancers could be episomal, integrated, or both. HPV integration could serve as a biomarker for clinical diagnosis, treatment, and prognosis. Although whole-genome sequencing by next-generation sequencing (NGS) technologies, such as the Illumina sequencing platform, have been used for detecting integrated HPV genome in cervical cancer, it faces challenges of analyzing long repeats and translocated sequences. In contrast, Oxford nanopore sequencing technology can generate ultra-long reads, which could be a very useful tool for determining HPV genome sequence and its physical status in cervical cancer. As a proof of concept, in this study, we completed whole genome sequencing from a cervical cancer tissue and a CaSki cell line with Oxford Nanopore Technologies. From the cervical cancer tissue, a 7,894 bp-long HPV35 genomic sequence was assembled from 678 reads at 97-fold coverage of HPV genome, sharing 99.96% identity with the HPV sequence obtained by Sanger sequencing. A 7904 bp-long HPV16 genomic sequence was assembled from data generated from the CaSki cell line at 3857-fold coverage, sharing 99.99% identity with the reference genome (NCBI: U89348). Intriguingly, long reads generated by nanopore sequencing directly revealed chimeric cellular–viral sequences and concatemeric genomic sequences, leading to the discovery of 448 unique integration breakpoints in the CaSki cell line and 60 breakpoints in the cervical cancer sample. Taken together, nanopore sequencing is a unique tool to identify HPV sequences and would shed light on the physical status of HPV genome in its associated cancers.



2018 ◽  
Author(s):  
Ratna Veeramachaneni ◽  
Thomas Walker ◽  
Antoine De Weck ◽  
Timothée Revil ◽  
Dunarel Badescu ◽  
...  

AbstractHead and neck squamous cell carcinoma (HNSCC) is a widely prevalent cancer globally with high mortality and morbidity. We report here changes in the genomic landscape in the development of HNSCC from potentially premalignant lesions (PPOLS) to malignancy and lymph node metastases. Frequent likely pathological mutations are restricted to a relatively small set of genes includingTP53, CDKN2A,FBXW7,FAT1,NOTCH1andKMT2D; these arise early in tumour progression and are present in PPOLs withNOTCH1mutations restricted to cell lines from lesions that subsequently progressed to HNSCC. The most frequent genetic changes are of consistent somatic copy number alterations (SCNA). The earliest SCNAs involved deletions ofCSMD1(8p23.2),FHIT(3p14.2) andCDKN2A(9p21.3) together with gains of chromosome 20.CSMD1deletions or promoter hypermethylation were present in all of the immortal PPOLs and occurred at high frequency in the immortal HNSCC cell lines (promoter hypermethylation ~63%, hemizygous deletions ~75%, homozygous deletions ~18%). Forced expression ofCSMD1in the HNSCC cell line H103 showed significant suppression of proliferation (p=0.0053) and invasioninvitro(p=5.98X10−5) supporting a role forCSMD1inactivation in early head and neck carcinogenesis. In addition, knockdown ofCSMD1in theCSMD1-expressing BICR16 cell line showed significant stimulation of invasionin vitro(p=1.82 x 10−5) but not cell proliferation (p=0.239). HNSCC with and without nodal metastases showed some clear differences including high copy number gains ofCCND1, hsa-miR-548k andTP63in the metastases group. GISTIC peak SCNA regions showed significant enrichment (adj P<0.01) of genes in multiple KEGG cancer pathways at all stages with disruption of an increasing number of these involved in the progression to lymph node metastases. Sixty-seven genes from regions with statistically significant differences in SCNA/LOH frequency between immortal PPOL and HNSCC cell lines showed correlation with expression including 5 known cancer drivers.Lay SummaryCancers affecting the head and neck region are relatively common. A large percentage of these are of one particular type; these are generally detected late and are associated with poor prognosis. Early detection and treatment dramatically improve survival and reduces the damage associated with the cancer and its treatment. Cancers arise and progress because of changes in the genetic material of the cells. This study focused on identifying such changes in these cancers particularly in the early stages of development, which are not fully known. Identification of these changes is important in developing new treatments as well as markers of behaviour of cancers and also the early or ‘premalignant’ lesions. We used a well-characterised panel of cell lines generated from premalignant lesions as well as cancers, to identify mutations in genes, and an increase or decrease in number of copies of genes. We mapped new and previously identified changes in these cancers to specific stages in the development of these cancers and their spread. We additionally report here for the first time, alterations inCSMD1gene in early premalignant lesions; we further show that this is likely to result in increased ability of the cells to spread and possibly, multiply faster as well.



2021 ◽  
Vol 11 ◽  
Author(s):  
May Poh Yik Goh ◽  
Norhayati Ahmad ◽  
Hartini Yasin ◽  
Abdalla Jama

Background: Dillenia suffruticosa (Griff.) Mart. has been traditionally used to promote wound healing, relieve rheumatism, fever and some cancerous growths. The leaves of the local variety of D. suffruticosa lack scientific studies on its biological applications in the context of antibacterial, antioxidant and cytotoxic activities. Objective: To evaluate the antioxidant, antibacterial and cytotoxic properties of the leaves of D. suffruticosa from Brunei Darussalam. Methods: The leaves were extracted using 80% (v/v) methanol, 80% (v/v) ethanol and aqueous. The antioxidant capacities were determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, 2,2’-azino-bis(3-ethylbenzothia zoline-6-sulfonic acid) (ABTS) radical scavenging and ferric ion reducing antioxidant power (FRAP) assays. The FolinCiocalteu and aluminium chloride colorimetric assays were also used to evaluate the total phenolic and flavonoid contents. The antibacterial and cytotoxic activities of the extracts were determined using the Kirby-Bauer disc diffusion and MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell proliferation assays. Results: The methanolic extract of the D. suffruticosa leaves displayed the highest antioxidant activity despite having comparable phenol content when extracted using the ethanol extraction solvent. The methanolic extract also demonstrated antibacterial activity on Staphylococcus aureus at a concentration of 50 mg/mL or above. The cytotoxicity of the methanolic extract was higher against the CaSki cell line than the A549 lung cancer cell line in the first 24 h but became more cytotoxic against A549 than CaSki at 48 h and 72 h. Conclusion: Our findings suggest that the methanolic extract of the leaves of D. suffruticosa from Brunei Darussalam has significant antioxidant and antibacterial activity against S. aureus and moderate cytotoxicity against A549 and CaSki cell lines.



2021 ◽  
Author(s):  
Serena Judith DSouza ◽  
Arati Mane ◽  
Linata Patil ◽  
Aazam Shaikh ◽  
Madhuri Thakar ◽  
...  

Abstract Background High Risk Human Papilloma Viruses (HR-HPV) recurrently infect women having Human Immunodeficiency Virus − 1 (HIV-1)infection. Transforming HPV E6 and E7 genes promote invasive cancers and interact with Notch-1receptor. Concomitantly, HIV-1 Tat binds to EGF motifs within the Notch-1extracellular domain. HR-HPV infection activates Notch-1 signalling. Permissive HIV-1entry into the cervix is allowed. Notch-1 inhibitors may offer solace to the aggressive cancer phenotype in HIV-1 positive women. Still, the molecular cross talk between different oncogenes within the Notch-1pathway during HIV-1/HPV-16 + co-infections has not been elucidated. Methods Adherent cervical tumor derived cell lines-CaSki cell line (with inherent HPV16+ sequence and endogenous Notch-1 activity) and C33A cell line (cervical cancer phenotype, HPV -ve for HPV DNA and RNA) were used. Plasmids (pLEGFPN1 encoding HIV-1 Tat and pNL 4 − 3 encoding HIV-1(full HIV-1 genome), were transfected at 600 ng/mL into the above mentioned cell lines, in order to examine independent effects of HIV-1 Tat and HIV-1 transfection in HPV-16+ cervical cancer cells. We performed western blotting, cell cycle analysis and RT-qPCR post transfection. Three sets of independent experiments were analyzed by Graph Pad Prism 5. Statistical significance was calculated using Student t -test. Data expressed as mean ± standard deviation (SD). p values ≤ 0.05* were considered statistically significant. Results HIV-1 Tat and HIV-1 inhibited Notch-1expression, with differential effects on EGFR when comparing C33A and CaSki cell lines. CDK2 induction in Tat transfected CaSki cells, showed concomitant G0/G1 phase accumulation favouring cancer progression. Notch-1 inhibition shut off significant Cyclin D expression with a significant p21 induction and increased G2-M cell population in CaSki cells. On the contrary, HIV-1 infection utilized Hes-1-EGFR-CyclinD-p21axis, G2-M arrest, DDR response and cancer progression. Conclusion Our study highlights for the first time that HIV-1 Tat and/or HIV-1 driven cancers in HPV-16+ CaSki cells, show Notch-1 suppression and CDK2 dependent activity. HIV-1 Tat activates Hes-1 amplifying the EGFR gene which improves the aggressive state possibly through irreversible oxidative-stress induced senescence. HIV-1, favours cancer progression through its CXCR4 receptor, responsible for unbridled mitosis, G2-M arrest and damaged DNA response (DDR). Treatment of CaSki cells with a Notch-1 inhibitor, DAPT showed marginal recovery in p21expression with Go/G1 and S phase recovery.



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