RNA Interference of Human Papillomavirus Type 18 E6 and E7 Induces Senescence in HeLa Cells

ABSTRACT The human papillomavirus oncoproteins E6 and E7 promote cell proliferation and contribute to carcinogenesis by interfering with the activities of cellular tumor suppressors. We used a small interfering RNA molecule targeting the E7 region of the bicistronic E6 and E7 mRNA to induce RNA interference, thereby reducing expression of E6 and E7 in HeLa cells. RNA interference of E6 and E7 also inhibited cellular DNA synthesis and induced morphological and biochemical changes characteristic of cellular senescence. These results demonstrate that reducing E6 and E7 expression is sufficient to cause HeLa cells to become senescent.

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Intratumor injection of small interfering RNA-targeting human papillomavirus 18 E6 and E7 successfully inhibits the growth of cervical cancer

International Journal of Oncology ◽

10.3892/ijo.29.3.541 ◽

2006 ◽

Cited By ~ 3

Author(s):

Takuma Fujii ◽

Miyuki Saito ◽

Eri Iwasaki ◽

Takahiro Ochiya ◽

Yoshifumi Takei ◽

...

Keyword(s):

Cervical Cancer ◽

Human Papillomavirus ◽

Small Interfering Rna ◽

Rna Targeting ◽

E6 And E7 ◽

Human Papillomavirus 18 ◽

Interfering Rna

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Faculty Opinions recommendation of DICER-LIKE 4 is required for RNA interference and produces the 21-nucleotide small interfering RNA component of the plant cell-to-cell silencing signal.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1029052.345437 ◽

2005 ◽

Author(s):

Andy Maule

Keyword(s):

Rna Interference ◽

Plant Cell ◽

Small Interfering Rna ◽

Interfering Rna

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Amphiphysin 1 Is Important for Actin Polymerization during Phagocytosis

Molecular Biology of the Cell ◽

10.1091/mbc.e07-04-0296 ◽

2007 ◽

Vol 18 (11) ◽

pp. 4669-4680 ◽

Cited By ~ 30

Author(s):

Hiroshi Yamada ◽

Emiko Ohashi ◽

Tadashi Abe ◽

Norihiro Kusumi ◽

Shun-AI Li ◽

...

Keyword(s):

Rna Interference ◽

Sertoli Cells ◽

Small Interfering Rna ◽

Actin Polymerization ◽

Actin Dynamics ◽

Polymerization Process ◽

Amphiphysin 1 ◽

Phagocytic Cups ◽

Interfering Rna ◽

Constitutively Active

Amphiphysin 1 is involved in clathrin-mediated endocytosis. In this study, we demonstrate that amphiphysin 1 is essential for cellular phagocytosis and that it is critical for actin polymerization. Phagocytosis in Sertoli cells was induced by stimulating phosphatidylserine receptors. This stimulation led to the formation of actin-rich structures, including ruffles, phagocytic cups, and phagosomes, all of which showed an accumulation of amphiphysin 1. Knocking out amphiphysin 1 by RNA interference in the cells resulted in the reduction of ruffle formation, actin polymerization, and phagocytosis. Phagocytosis was also drastically decreased in amph 1 (−/−) Sertoli cells. In addition, phosphatidylinositol-4,5-bisphosphate–induced actin polymerization was decreased in the knockout testis cytosol. The addition of recombinant amphiphysin 1 to the cytosol restored the polymerization process. Ruffle formation in small interfering RNA-treated cells was recovered by the expression of constitutively active Rac1, suggesting that amphiphysin 1 functions upstream of the protein. These findings support that amphiphysin 1 is important in the regulation of actin dynamics and that it is required for phagocytosis.

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HPV-18 E6 Oncoprotein and Its Spliced Isoform E6*I Regulate the Wnt/β-Catenin Cell Signaling Pathway through the TCF-4 Transcriptional Factor

International Journal of Molecular Sciences ◽

10.3390/ijms19103153 ◽

2018 ◽

Vol 19 (10) ◽

pp. 3153 ◽

Cited By ~ 6

Author(s):

J. Muñoz-Bello ◽

Leslie Olmedo-Nieva ◽

Leonardo Castro-Muñoz ◽

Joaquín Manzo-Merino ◽

Adriana Contreras-Paredes ◽

...

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Signaling Pathway ◽

Transcriptional Activation ◽

Target Genes ◽

Promote Cell Proliferation ◽

E6 And E7 ◽

Hpv 18

The Wnt/β-catenin signaling pathway regulates cell proliferation and differentiation and its aberrant activation in cervical cancer has been described. Persistent infection with high risk human papillomavirus (HR-HPV) is the most important factor for the development of this neoplasia, since E6 and E7 viral oncoproteins alter cellular processes, promoting cervical cancer development. A role of HPV-16 E6 in Wnt/β-catenin signaling has been proposed, although the participation of HPV-18 E6 has not been previously studied. The aim of this work was to investigate the participation of HPV-18 E6 and E6*I, in the regulation of the Wnt/β-catenin signaling pathway. Here, we show that E6 proteins up-regulate TCF-4 transcriptional activity and promote overexpression of Wnt target genes. In addition, it was demonstrated that E6 and E6*I bind to the TCF-4 (T cell factor 4) and β-catenin, impacting TCF-4 stabilization. We found that both E6 and E6*I proteins interact with the promoter of Sp5, in vitro and in vivo. Moreover, although differences in TCF-4 transcriptional activation were found among E6 intratype variants, no changes were observed in the levels of regulated genes. Furthermore, our data support that E6 proteins cooperate with β-catenin to promote cell proliferation.

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DrosophilaoncogeneGas41is an RNA interference modulator that intersects heterochromatin and the small interfering RNA pathway

FEBS Journal ◽

10.1111/febs.13115 ◽

2014 ◽

Vol 282 (1) ◽

pp. 153-173 ◽

Cited By ~ 4

Author(s):

Sumit G. Gandhi ◽

Indira Bag ◽

Saswati Sengupta ◽

Manika Pal-Bhadra ◽

Utpal Bhadra

Keyword(s):

Rna Interference ◽

Small Interfering Rna ◽

Interfering Rna

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Transfection of Hairpin Small Interfering RNA Expression Vector Targeting Rat Nuclear Factor (NF) (κB) Inhibits Rat Cell Proliferation Induced by NF-κB Signal Pathway Activation

Transplantation Proceedings ◽

10.1016/j.transproceed.2010.09.168 ◽

2010 ◽

Vol 42 (10) ◽

pp. 4633-4637 ◽

Cited By ~ 2

Author(s):

X.P. He ◽

X.X. Li ◽

Z.H. Wang ◽

Y.W. Bi ◽

F.Y. Liu ◽

...

Keyword(s):

Cell Proliferation ◽

Small Interfering Rna ◽

Expression Vector ◽

Signal Pathway ◽

Rna Expression ◽

Nuclear Factor ◽

Pathway Activation ◽

Vector Targeting ◽

Interfering Rna

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Structurally modulated codelivery of siRNA and Argonaute 2 for enhanced RNA interference

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1719565115 ◽

2018 ◽

Vol 115 (12) ◽

pp. E2696-E2705 ◽

Cited By ~ 20

Author(s):

Jiahe Li ◽

Connie Wu ◽

Wade Wang ◽

Yanpu He ◽

Elad Elkayam ◽

...

Keyword(s):

Molecular Structure ◽

Rna Interference ◽

Small Interfering Rna ◽

Sirna Delivery ◽

Effector Protein ◽

Side Group ◽

Argonaute 2 ◽

Fundamental Research ◽

Delivery Vehicles ◽

Interfering Rna

Small interfering RNA (siRNA) represents a promising class of inhibitors in both fundamental research and the clinic. Numerous delivery vehicles have been developed to facilitate siRNA delivery. Nevertheless, achieving highly potent RNA interference (RNAi) toward clinical translation requires efficient formation of RNA-induced gene-silencing complex (RISC) in the cytoplasm. Here we coencapsulate siRNA and the central RNAi effector protein Argonaute 2 (Ago2) via different delivery carriers as a platform to augment RNAi. The physical clustering between siRNA and Ago2 is found to be indispensable for enhanced RNAi. Moreover, by utilizing polyamines bearing the same backbone but distinct cationic side-group arrangements of ethylene diamine repeats as the delivery vehicles, we find that the molecular structure of these polyamines modulates the degree of siRNA/Ago2-mediated improvement of RNAi. We apply this strategy to silence the oncogene STAT3 and significantly prolong survival in mice challenged with melanoma. Our findings suggest a paradigm for RNAi via the synergistic coassembly of RNA with helper proteins.

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