Partial silencing of fucosyltransferase 8 gene expression inhibits proliferation of Ishikawa cells, a cell line of endometrial cancer

Objective: The present study aimed to explore the function of sirtuin 2 (SIRT2) on cell proliferation, apoptosis, rat sarcoma virus (RAS)/ extracellular signal-regulated kinase (ERK) pathway in endometrial cancer (EC). Methods: SIRT2 expression in human EC cell lines and human endometrial (uterine) epithelial cell (HEEC) line was assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot. SIRT2 knock-down and control knock-down plasmids were transfected into HEC1A cells, respectively; SIRT2 overexpression and control overexpression plasmids were transfected into Ishikawa cells, respectively. After transfection, SIRT2, HRas proto-oncogene, GTPase (HRAS) expressions were evaluated by RT-qPCR and western blot. ERK and phosphorylated ERK (pERK) expressions were evaluated by western blot. Meanwhile, cell proliferation and cell apoptosis were measured. Results: Compared to normal HEEC cell line, SIRT2 mRNA and protein expressions were increased in most human EC cell lines (including HEC1A, RL952 and AN3CA), while were similar in Ishikawa cell line. In HEC1A cells, SIRT2 knock-down decreased cell proliferation but increased apoptosis. In Ishikawa cells, SIRT2 overexpression induced cell proliferation but inhibited apoptosis. For RAS/ERK pathway, SIRT2 knock-down reduced HRAS and inactivated pERK in HEC1A cells, whereas SIRT2 overexpression increased HRAS and activated pERK in Ishikawa cells, suggesting that SIRT2 was implicated in the regulation of RAS/ERK pathway in EC cells. Conclusion: SIRT2 contributes to the EC tumorigenesis, which appears as a potential therapeutic target.

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Derivation and Characterization of a ES-Like Cell Line from Indian CatfishHeteropneustes fossilisBlastulas

The Scientific World JOURNAL ◽

10.1155/2014/427497 ◽

2014 ◽

Vol 2014 ◽

pp. 1-9 ◽

Cited By ~ 2

Author(s):

Anindya S. Barman ◽

Kuldeep K. Lal ◽

Gaurav Rathore ◽

Vindhya Mohindra ◽

Rajeev K. Singh ◽

...

Keyword(s):

Gene Expression ◽

Cell Line ◽

Es Cells ◽

Embryonic Cells ◽

Dna Profile ◽

Expression Studies ◽

C Subunit ◽

Gfp Reporter Gene ◽

A Cell ◽

Gene Expression Studies

A cell line designated as HFB-ES was established from blastula stage embryos ofH. fossilis(Singhi). The embryonic cells were harvested and maintained in Leibovitz’s medium supplemented with 15% fetal bovine serum. The cell line had been subcultured for more than 90 passages in a period of 24 months. HFB-ES cells were able to grow at temperatures between 25 and 35°C with an optimum temperature of 28°C. The growth rate of HFB-ES was proportional to FBS concentration, with optimum growth seen at 15% FBS concentration. The originality of the cell line was confirmed by sequencing of cytochrome oxidase c subunit I (COI), cytochrome b gene, and microsatellite DNA profile. Results of chromosome complements of HFB showed normal karyo-morphology with 56 (2n) diploid number of chromosomes after 40 passages which indicated that the developed cell line is chromosomally stable. The pluripotency of HFB was demonstrated by alkaline phosphatase activity and Oct-4 gene expression. Expression of GFP reporter gene was successful in HFB-ES. These results indicated that HFB-ES could be utilized for future gene expression studies.

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KLE: A cell line with defective estrogen receptor derived from undifferentiated endometrial cancer

Gynecologic Oncology ◽

10.1016/0090-8258(84)90080-5 ◽

1984 ◽

Vol 17 (2) ◽

pp. 213-230 ◽

Cited By ~ 50

Author(s):

George S. Richardson ◽

G.Richard Dickersin ◽

Leonard Atkins ◽

David T. MacLaughlin ◽

Shanthi Raam ◽

...

Keyword(s):

Estrogen Receptor ◽

Endometrial Cancer ◽

Cell Line ◽

A Cell

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Visualizing Gene Expression in Real-Time

Microscopy Today ◽

10.1017/s1551929500051567 ◽

2005 ◽

Vol 13 (3) ◽

pp. 3-7

Author(s):

Stephen W. Carmichael

Keyword(s):

Gene Expression ◽

Cell Line ◽

Real Time ◽

Temporal Resolution ◽

Protein Sequence ◽

Living Cells ◽

Entire Process ◽

Gene Chips ◽

A Cell ◽

Dna Template

Gene expression has been visualized for a few decades, but in static forms such as blots and gene chips. Susan Janicki, Toshiro Tsukamoto, Sim one Salghetti, William Tansey, Ravi Sachidanandam, Kannanganattu Prasanth, Thomas Ried, Yaron Shav-Tal, Edouard Bertrand, Robert Singer, and David Spector have recently designed a cell line in which gene expression can be observed with stunningly accurate spatial and temporal resolution. Gene expression is a cascade of events beginning with transcription of RNA from the DNA template and ending with translation into a protein sequence. Janicki et al. were able to visualize the entire process at the levels of DNA, RNA, and proteins in living cells!

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Development of Caudal Fin Cell Line from Hill Trout Barilius Bendelisis (Hamilton, 1807) for Cytotoxicity and Transfection Studies

Croatian Journal of Fisheries ◽

10.2478/cjf-2021-0002 ◽

2021 ◽

Vol 79 (1) ◽

pp. 15-24

Author(s):

Murali Sanjeev Kumar ◽

Pankaj Soni ◽

Ravindra Kumar ◽

Neha Singh ◽

Shreya Srivastava ◽

...

Keyword(s):

Gene Expression ◽

Cell Line ◽

Mitochondrial Gene ◽

Toxicity Screening ◽

Green Fluorescence ◽

Caudal Fin ◽

A Cell ◽

Barilius Bendelisis ◽

Early Toxicity ◽

N Vector

Abstract A cell line named BBdF-1, established from the caudal fin of hill stream fish Barilius bendelisis, has been subcultured for more than 52 passages and is being maintained in L-15 media containing 20% FBS. Species origin of the cell line was confirmed using amplification of partial region of 16S and COI mitochondrial gene sequences. The optimum temperature for growth of BBdF-1 cell line was found to be 28°C. Karyotyping revealed diploid chromosome number as 50. Cells exhibited strong binding for cytokeratin marker and thus were found to be epithelial-like. Strong green fluorescence was observed in BBdF-1 cells transfected with phrGFP-II-N vector, indicating its suitability for utilization in gene expression and manipulation studies. Successful assessment of cytotoxicity of two heavy metals, viz. mercury and chromium, was performed. The cell line can serve as a useful resource material for early toxicity screening of pesticides/pollutant and gene expression.

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In Vivo and In Vitro Analysis of Baculovirusie-2 Mutants

Journal of Virology ◽

10.1128/jvi.73.3.2460-2468.1999 ◽

1999 ◽

Vol 73 (3) ◽

pp. 2460-2468 ◽

Cited By ~ 40

Author(s):

Elena A. Prikhod’ko ◽

Albert Lu ◽

Joyce A. Wilson ◽

Lois K. Miller

Keyword(s):

Gene Expression ◽

Cell Line ◽

Late Gene ◽

Wild Type ◽

Late Gene Expression ◽

Specific Effects ◽

Viral Promoters ◽

Occlusion Bodies ◽

A Cell ◽

Budded Virus

ABSTRACT Upon transient expression in cell culture, the ie-2gene of Autographa californica nuclear polyhedrosis virus (AcMNPV) displays three functions: transactivation of viral promoters, direct or indirect stimulation of virus origin-specific DNA replication, and arrest of the cell cycle. The ability of IE2 to trans stimulate DNA replication and coupled late gene expression is observed in a cell line derived fromSpodoptera frugiperda but not in a cell line derived fromTrichoplusia ni. This finding suggested that IE-2 may exert cell line-specific or host-specific effects. To examine the role of ie-2 in the context of infection and its possible influence on the host range, we constructed recombinants of AcMNPV containing deletions of different functional regions within ie-2 and characterized them in cell lines and larvae of S. frugiperda and T. ni. Theie-2 mutant viruses exhibited delays in viral DNA synthesis, late gene expression, budded virus production, and occlusion body formation in SF-21 cells but not in TN-5B1-4 cells. In TN-5B1-4 cells, the ie-2 mutants produced more budded virus and fewer occlusion bodies but the infection proceeded without delay. Examination of the effects of ie-2 and the respective mutants on immediate-early viral promoters in transient expression assays revealed striking differences in the relative levels of expression and differences in responses to ie-2 and its mutant forms in different cell lines. In T. ni and S. frugiperda larvae, the infectivities of the occluded form ofie-2 mutant viruses by the normal oral route of infection was 100- and 1,000-fold lower, respectively, than that of wild-type AcMNPV. The reduction in oral infectivity was traced to the absence of virions within the occlusion bodies. The infectivity of the budded form of ie-2 mutants by hemocoelic injection was similar to that of wild-type virus in both species. Thus,ie-2 mutants are viable but exhibit cell line-specific effects on temporal regulation of the infection process. Due to its effect on virion occlusion, mutants of IE-2 were essentially noninfectious by the normal route of infection in both species tested. However, since budded viruses exhibited normal infectivity upon hemocoelic injection, we conclude that ie-2 does not affect host range per se. The possibility that IE-2 exerts tissue-specific effects has not been ruled out.

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Hydroethanolic Extracts of the Aristotelia Chilensis (Maqui) Berry Reduces Cellular Viability and Invasiveness in the Endometrial Cancer Cell Line Ishikawa

Integrative Cancer Therapies ◽

10.1177/15347354211007560 ◽

2021 ◽

Vol 20 ◽

pp. 153473542110075

Author(s):

Javier Mena ◽

Estefanía Elgueta ◽

Francisca Espinola-Gonzales ◽

Hugo Cardenas ◽

Pedro A. Orihuela

Keyword(s):

Endometrial Cancer ◽

Cell Viability ◽

Cell Line ◽

Cancer Cell ◽

Cancer Cell Line ◽

Boyden Chamber ◽

Female Population ◽

Endometrial Cancer Cell ◽

Ishikawa Cells ◽

Aristotelia Chilensis

Cancer of the reproductive tract includes diseases with higher prevalence in the female population. This investigation examined whether an anthocyanin-enriched extract of Aristotelia chilensis, commonly known as “maqui,” could affect some hallmarks of endometrial cancer. Cultures of the human endometrial cancer cell line Ishikawa were treated with a hydroethanolic maqui extract at 1, 3, 10, 30, 100, 300, or 1000 µg/mL to determine the effect on cell viability by MTT assay. Then, we used the 50% Effective Concentration (EC50) to evaluate whether the effect of the maqui extract is mediated via an arrest of the cell cycle or induction of apoptosis using flow cytometry or Annexin V-FITC assays, respectively. The effects of sublethal doses of the maqui extract on migration and invasiveness of Ishikawa cells were also evaluated by the wound healing and Boyden Chamber assay, respectively. Our results show that the hydroethanolic maqui extract inhibits the cell viability with an EC50 of 472.3 µg/mL via increased apoptosis, and that reduces the invasive capacity but not migration of Ishikawa cells. These findings suggest that the hydroethanolic maqui extract has antineoplastic properties for endometrial cancer and merits further studies to corroborate its efficiency as anticancer therapy in reproductive organs.

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The Establishment of Bortezomib Resistant Myeloma Cell Line KM3/BTZ and Explore the Resistance Mechanism

Blood ◽

10.1182/blood.v124.21.5226.5226 ◽

2014 ◽

Vol 124 (21) ◽

pp. 5226-5226 ◽

Cited By ~ 2

Author(s):

Luqun Wang ◽

Qiong Liu ◽

Hao Li ◽

Li Lizhen ◽

Xin Wang

Keyword(s):

Gene Expression ◽

Multiple Myeloma ◽

Cell Line ◽

Resistance Index ◽

Resistant Cell ◽

Resistant Cell Line ◽

Rt Pcr ◽

A Cell ◽

Family Gene ◽

Mdr 1

Abstract Background: Multiple myeloma (MM) is an incurable B-cell malignancy resulting in significant morbidity and mortality, the incidence of second place in hematological malignancies. The proteasome inhibitor bortezomib inhibits IkappaB degradation, prevents NF-kappaB activation, and induces apoptosis in MM cells, has become first-line drug to MM. Despite its promising activity in traditional chemoresistant myeloma patients, however, some patients are resistant or become refractory to bortezomib. Chauhan D et al reported that Blockade of Hsp27 overcomes bortezomib resistance in lymphoma cells (Chauhan D et al., Cancer Re, 2003). However, bortezomib resistance mechanisms in MM remain controversial, the molecular basis of this reduced responsiveness is currently not fully understood. Objective: To establish a bortezomib-resistant cell line KM3 /BTZ of MM, then search for genes related to bortezomib resistance phenotype by analyzing the differential gene expression patterns with cDNA microarray,and explain the mechanism of bortezomib-resistant multiple myeloma. Methods: Bortezomib-resistant KM3/BTZ cell line was generated by increasing concentration of bortezomib to induce KM3 cells in vitro. MTT assay was employed to detect the cytotoxic effect on KM3 and KM3/BTZ cells. The cells apoptosis were analyzed by flow cytometry. We detect the gene expression profile changes between KM3/BTZ and its parent cell line KM3 cells by CDNA microarray. Combining of molecular annotation system MAS3.0 software and detailed analysis of documented resistance genes were used to analyze the data. Specific differentially-expressed genes were chosen for further verification using real-time RT-PCR. The expression of multi-drug resistance 1(MDR-1) gene mRNA was determined by RT-PCR. Results: The KM3/BTZ cell line was established successfully. Bortezomib resistance index values were significantly higher in KM3/BTZ (IC50 =351.2±3.51ng /ml) when compared to KM3 cells (IC50 = 17.8±1.03ng /ml),and the resistance index is 19.7 (P< 0.05). Through gene expression profiling filtering, compared with KM3,770 significantly differently expressed genes were screened out in the KM3/BTZ cell line, of which up and down regulated genes were 287 and 383 respectively, in which heat shock protein family gene HSPB2 was significantly up-regulated (Ratio=14.7455). Downregulated gene list in KM3/BTZ cell line included transcriptional regulators like ZNF family proteins (Tab 1). Upregulated genes in KM3/BTZ cell line included signal transduction related genes like MS4A family protein (Tab 1) also transcriptional regulators like ZNF family proteins (Tab 1). The expression trend of eight genes for further verification was almost consistent with the microarray, excepting gene of JUN (Fig 1). The expression of MDR-1 mRNA was not observed in either resistant or parental cells (Fig 2). Conclusion: We have successfully established bortezomib-resistant cell line KM3/BTZ. MDR-1 was not involved in bortezomib-resistant multiple myeloma cells.HSPB2, ZNF and MS4A family genes are probably related to bortezomib resistance in KM3/BTZ cells. Our results suggest that they may be new targets to overcome resistance to bortezomib in patients with MM. Table 1. ZNF and MS4A family gene expression in KM3/BTZ and KM3 cells Gene B signal* B detection* A signal* A detection* BvsA signal log ratio* BvsA change* ZNF711 85.08 P 206.16 P 0.41269 D ZNF92 6.75 P 14.57 P 0.46328 D ZNF704 10.7 A 26.78 P 0.39955 D ZNF492 7.08 P 3.48 P 2.03448 I ZNF532 87.93 P 43.54 A 2.01952 I ZNF594 10.25 P 4.63 P 2.21382 I ZNF789 269.79 P 97.3 P 2.77276 I ZNF506 330.36 P 149.48 P 2.21006 I MS4A3 1721.58 P 145.3 P 11.84845 I MS4A4A 2688.23 P 100.8 P 26.66895 I MS4A6A 141.96 P 56.47 M 2.513901 I MS4A1 49.02 P 53.11 P 0.92299 NC P: High expression; A: Low expression; M:Marginally D: Decrease; I: Increase; NC: No change *: A:KM3; B: KM3/BTZ Figure 1 Figure 1. Correlations of gene expression changes between oligonueleotide Microarray and qRT-PCR, showing change of selected gene in KM3/BTZ compared with KM3.The Y-axis indicates the log2 transformed ratio of mRNA expression. Figure 2 Figure 2. The expression level of MDR-1 mRNA in KM3 and KM3/BTZ and K562/A cell lines. M: DNA Marker; 1-2: K562/A cell line; 3-4: KM3cell line; 5-6: KM3/BTZ cell line Disclosures No relevant conflicts of interest to declare.

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Ribozyme-mediated inhibition of bcr-abl gene expression in a Philadelphia chromosome-positive cell line

Blood ◽

10.1182/blood.v82.2.600.600 ◽

1993 ◽

Vol 82 (2) ◽

pp. 600-605 ◽

Cited By ~ 104

Author(s):

DS Snyder ◽

Y Wu ◽

JL Wang ◽

JJ Rossi ◽

P Swiderski ◽

...

Keyword(s):

Gene Expression ◽

Cell Line ◽

Cell Biology ◽

Fusion Gene ◽

Therapeutic Potential ◽

Blast Crisis ◽

Philadelphia Chromosome ◽

Myelogenous Leukemia ◽

A Cell

Abstract The bcr-abl fusion gene is the molecular counterpart of the Philadelphia chromosome (Ph1) and is directly involved in the pathogenesis of Ph1+ leukemia. Inhibition of bcr-abl gene expression may have profound effects on the cell biology of Ph1+ cells, as recent experiments with antisense oligonucleotides have shown. In this study we have designed and synthesized a unique ribozyme that is directed against bcr-abl mRNA. The ribozyme cleaved bcr-abl mRNA in a cell-free in vitro system. A DNA-RNA hybrid ribozyme was then incorporated into a liposome vector and transfected into EM-2 cells, a cell line derived from a patient with blast crisis of chronic myelogenous leukemia. The ribozyme decreased levels of detectable bcr-abl mRNA in these cells, inhibited expression of the bcr-abl gene product, p210bcr-abl, and inhibited cell growth. This anti-bcr-abl ribozyme may be a useful tool to study the cell biology of Ph1+ leukemia and may ultimately have therapeutic potential in treating patients with Ph1 leukemias.

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Gene expression effects of lithium and valproic acid in a serotonergic cell line

Physiological Genomics ◽

10.1152/physiolgenomics.00069.2018 ◽

2019 ◽

Vol 51 (2) ◽

pp. 43-50 ◽

Cited By ~ 5

Author(s):

Diana Balasubramanian ◽

John F. Pearson ◽

Martin A. Kennedy

Keyword(s):

Gene Expression ◽

Valproic Acid ◽

Cell Line ◽

Mood Stabilizer ◽

Underlying Mechanism ◽

Matrix Remodeling ◽

Rna Seq ◽

Significant Enrichment ◽

A Cell ◽

Serotonergic Cell

Valproic acid (VPA) and lithium are widely used in the treatment of bipolar disorder. However, the underlying mechanism of action of these drugs is not clearly understood. We used RNA-Seq analysis to examine the global profile of gene expression in a rat serotonergic cell line (RN46A) after exposure to these two mood stabilizer drugs. Numerous genes were differentially regulated in response to VPA (log2 fold change ≥ 1.0; i.e., odds ratio of ≥2, at false discovery rate <5%), but only two genes ( Dynlrb2 and Cdyl2) showed significant differential regulation after exposure of the cells to lithium, with the same analysis criteria. Both of these genes were also regulated by VPA. Many of the differentially expressed genes had functions of potential relevance to mood disorders or their treatment, such as several serpin family genes (including neuroserpin), Nts (neurotensin), Maob (monoamine oxidase B), and Ap2b1, which is important for synaptic vesicle function. Pathway analysis revealed significant enrichment of Gene Ontology terms such as extracellular matrix remodeling, cell adhesion, and chemotaxis. This study in a cell line derived from the raphe nucleus has identified a range of genes and pathways that provide novel insights into potential therapeutic actions of the commonly used mood stabilizer drugs.

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