Effect of RNA interference targeting human telomerase reverse transcriptase on telomerase and its related protein expression in nasopharyngeal carcinoma cells

2006 ◽  
Vol 121 (5) ◽  
pp. 476-482 ◽  
Author(s):  
Yan Wang ◽  
Hong-Gang Duan ◽  
Shi-Ming Chen ◽  
Bo-Kui Xiao ◽  
Jie Cheng ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Proliferating Cell Nuclear Antigen ◽  
Short Hairpin Rna ◽  
Nuclear Antigen ◽  
Telomerase Reverse Transcriptase ◽  
Hairpin Rna ◽  
Human Telomerase Reverse Transcriptase ◽  
Short Hairpin ◽  
Human Telomerase ◽  
Proliferating Cell

Objective: Analysis of the correlation between telomerase and the expression of its related proteins may provide insight into the molecular mechanism of nasopharyngeal carcinogenesis. We investigated the effect of short hair pin ribonucleic acid (RNA) specific for human telomerase reverse transcriptase messenger RNA on the expression of the proteins c-myc (the transcription factor c-myc is a shortlived nuclear phospho-protein involved in cell proliferation and differentiation, belongs to the myc family), proliferating cell nuclear antigen and Caspase-3 in nasopharyngeal carcinoma cells.Methods: Short hairpin RNA expression vectors targeting the messenger RNA of human telomerase reverse transcriptase were constructed. Cells were treated with the short hairpin RNA expression vectors targeting human telomerase reverse transcriptase or vectors that included mismatched short hairpin RNA, and telomerase activity was measured by telomeric repeat amplification enzyme-linked immunosorbent assay. Cell viability was examined using the 3-(4,5-dimethyl thizol-2-yl) 2,5-diphenyl tetrazolium bromide assay. The expression of the three proteins (c-myc, proliferating cell nuclear antigen and Caspase-3) was determined by Western blotting.Results: Short hairpin RNA specific for human telomerase reverse transcriptase messenger RNA significantly inhibited telomerase activity. In addition, the expression of and proliferating cell nuclear antigen were both inhibited, while the expression of Caspase-3 was up-regulated.Conclusions: Our results suggest that short hairpin RNA directed against human telomerase reverse transcriptase inhibits cell viability by regulating telomerase activity and its related proteins expression in nasopharyngeal carcinoma cells. Therefore, RNA interference technology may be a promising strategy for the treatment of nasopharyngeal cancer.

scholarly journals Total Flavones Isolated from Lycium Barbarum L. Inhibit the Proliferation, Migration and Invasion of GlioblastomaU-87MG Cells by Decreasing Formyl Peptide Receptor 1 Expression

2021 ◽  
Author(s):  
Yuankui Chu ◽  
Qing Wang ◽  
Yuan Li ◽  
Yating Zhu ◽  
Baibin Pang ◽  
...  
Keyword(s):  
Short Hairpin Rna ◽  
Formyl Peptide Receptor ◽  
Nuclear Antigen ◽  
Migration And Invasion ◽  
Dependent Manner ◽  
Lycium Barbarum ◽  
Hairpin Rna ◽  
Formyl Peptide ◽  
Short Hairpin ◽  
Proliferating Cell

Abstract Background: To study the role and mechanisms of total flavones from Lycium barbarum L. (TFL) and FPR1 in the growth of glioblastoma U-87MG cells.Main Methods: CCK-8, wound-healing and Transwell were used for investigating proliferation, motility and invasion of U87 cells after treating with total flavones. RT-qPCR and Western blot were used to study the effect of total flavones on proliferating cell nuclear antigen (PCNA), matrix metalloproteinase2 (MMP2) and FPR1. The short hairpin RNA and FPR agonist fMLP were used to delineate the role of FPR1.Results: TFL was successfully isolated, and its concentration was determined to be 6.205 mg/l. TFL inhibited the proliferation, migration and invasion of U-87MG cells in a time and dose-dependent manner compared to controls. Decreasing FPR1 expression using short hairpin RNA significantly inhibited the migration and invasion of U-87MG cells. Notably, increased expression of FPR1 and treatment with FPR-agonist peptides such as N-formylmethionyl-leucyl-phenylalanine induced the migration and invasion of U-87MG cells, which was significantly decreased when the cells were treated with TFL. Conclusion: TFL inhibits the proliferation, migration and invasion of human glioblastoma U-87MG cells through decreasing the expression of FPR1. These findings provide valuable evidence for the development of antitumor drugs.

scholarly journals Latency-Associated Nuclear Antigen of Kaposi's Sarcoma-Associated Herpesvirus Up-Regulates Transcription of Human Telomerase Reverse Transcriptase Promoter through Interaction with Transcription Factor Sp1

2004 ◽  
Vol 78 (19) ◽  
pp. 10348-10359 ◽  
Author(s):  
Subhash C. Verma ◽  
Sumit Borah ◽  
Erle S. Robertson
Keyword(s):  
Reverse Transcriptase ◽  
Kaposi's Sarcoma ◽  
Promoter Activity ◽  
Human Cells ◽  
Nuclear Antigen ◽  
Telomerase Reverse Transcriptase ◽  
Htert Promoter ◽  
Human Telomerase Reverse Transcriptase ◽  
Infected Cells ◽  
Human Telomerase

ABSTRACT Telomerase is required for the maintenance of telomere length and is an important determinant for cell immortalization. In human cells, telomerase activity is due to the expression of its enzymatic subunit, human telomerase reverse transcriptase (hTERT). The expression of hTERT is not typically detectable in healthy somatic human cells but is present in cancerous tissues and immortalized cells. We have previously shown that hTERT promoter activity is up-regulated by the Kaposi's sarcoma-associated herpesvirus (KSHV)-encoded latency-associated nuclear antigen (LANA). LANA is expressed in all forms of human malignancies associated with KSHV. The hTERT promoter sequence located at positions −130 to +5 contains several Sp1 binding motifs and was shown be important for up-regulation by LANA. In this report, we demonstrate that hTERT promoter activity is due to the direct interaction of LANA with Sp1. The interaction of LANA with Sp1 was demonstrated through in vitro binding experiments and coimmunoprecipitation and is supported by the colocalization of these two molecules in the nuclei of KSHV-infected cells. Moreover, LANA modulates Sp1-mediated transcription in transient GAL4 fusion reporter assays. Mapping of the regions involved in binding and transcriptional activation showed that the amino terminus of LANA is the major site for interaction and up-regulation but that it can cooperate with the carboxy terminus to enhance these functions. An analysis of Sp1 binding to its cognate sequence corroborated the binding data. Together, our results suggest that the interaction of LANA with Sp1 up-regulates the telomerase promoter, potentially contributing to the immortalization of KSHV-infected cells.

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