Highly potent and specific siRNAs against E6 or E7 genes of HPV16- or HPV18-infected cervical cancers

Chang, Jeng-Shou; Kuo, T-F; Yj, Chen; Chiu, Cheng-Tang; Lu, Yao; Li, H-F; Shen, Chia-Rui; Cheng, Ann‐Joy

doi:10.1038/cgt.2010.38

Cited by 81 publications

(69 citation statements)

References 21 publications

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“…Detailed information on cell lines is provided in Supplementary Table S4 and Supplementary Fig. S4. RNA extraction, cDNA generation, and quantitative PCR Cellular RNA isolation using the Nucleospin RNA II Kit (Macherey-Nagel) and cDNA generation was previously described (34). cDNA samples were subjected to real-time quantitative PCR using RT2 SYBR green master mix (SA Biosciences; ref.…”

Section: Cell Linesmentioning

confidence: 99%

“…Four siRNA oligonucleotides were synthesized by Dharmacon, including siPy2-X (which targets the 3 0 -UTR sequence of hPygo2 of endogenous hPygo2 mRNA), siPy2-Z (which targets the coding region of hPygo2), siE7 (34), and a nonspecific negative control, siNTC. We have previously described the Elf-1 targeting siRNA, siElf-1, (7).…”

Section: Rna Interference and Rescue Assaysmentioning

confidence: 99%

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Human Papilloma Virus (HPV) E7-Mediated Attenuation of Retinoblastoma (Rb) Induces hPygopus2 Expression via Elf-1 in Cervical Cancer

Tzenov

Andrews

Voisey

et al. 2013

Molecular Cancer Research

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The human papillomavirus (HPV) is the etiologic agent of cervical cancer. In this study, we provide evidence for the human Pygopus (hPygo)2 gene as a cellular biomarker for HPV-related disease. In a tumor microarray of cervical cancer progression, hPygo2 levels were greater in high-grade lesions and squamous cell carcinomas than in normal epithelia. Similarly, hPygo2 mRNA and protein levels were greater in HPV-positive cervical cancer cells relative to uninfected primary cells. RNA interference (RNAi)-mediated depletion of HPV-E7 increased whereas E74-like factor (Elf)-1 RNAi decreased association of Retinoblastoma (Rb) tumor suppressor with the hPygo2 promoter in cervical cancer cell lines. Transfection of dominant-active Rb inhibited Elf-1-dependent activation of hPygo2, whereas Elf-1 itself increased hPygo2 expression. Chromatin immunoprecipitation assays showed that Rb repressed hPygo2 by inhibiting Elf-1 at the Ets-binding site in the hPygo2 promoter. These results suggested that abrogation of Rb by E7 resulted in derepression of Elf-1, which in turn stimulated expression of hPygo2.

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Section: Cell Linesmentioning

confidence: 99%

Section: Rna Interference and Rescue Assaysmentioning

confidence: 99%

Human Papilloma Virus (HPV) E7-Mediated Attenuation of Retinoblastoma (Rb) Induces hPygopus2 Expression via Elf-1 in Cervical Cancer

Tzenov

Andrews

Voisey

et al. 2013

Molecular Cancer Research

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“…In recent years, there have been a number of publications showing the potential use of RNAi as a treatment for cervical cancer. [14][15][16][22][23][24][25] We demonstrated that RNAi triggered by short-hairpin RNA (shRNA) targeting a specific site within E7 mRNA could even induce immunity to E7 in immune-competent mice. 26 All the data indicate that RNAi-based therapy can be developed as a promising treatment for cervical cancer.…”

Section: Introductionmentioning

confidence: 99%

Silencing oncogene expression in cervical cancer stem-like cells inhibits their cell growth and self-renewal ability

Yeo

McMillan

et al. 2011

Cancer Gene Ther

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Accumulating evidence supports the concept that cancer stem cells (CSCs) are responsible for tumor initiation and maintenance. They are also considered as an attractive target for advanced cancer therapy. Using a sphere culture method that favors the growth of self-renewal cells, we have isolated sphere-forming cells (SFCs) from cervical cancer cell lines HeLa and SiHa. HeLa-SFCs were resistant to multiple chemotherapeutic drugs and were more tumorigenic, as evidenced by the growth of tumors following injection of immunodeficient mice with 1 Â 10 4 cells, compared with 1 Â 10 6 parental HeLa cells required to grow tumors of similar size in the same time frame. These cells showed an expression pattern of CD44 high /CD24 low that resembles the CSC surface biomarker of breast cancer. We further demonstrated that HeLa-SFCs expressed a higher level (6.9-fold) of the human papillomavirus oncogene E6, compared with that of parental HeLa cells. Gene silencing of E6 with a lentiviral-short-hairpin RNA (shRNA) profoundly inhibited HeLa-SFC sphere formation and cell growth. The inhibition of cell growth was even greater than that for sphere formation after E6 silence, suggesting that the loss of self-renewing ability may be more important. We then measured the expression of selfrenewal genes, transformation growth factor-beta (TGF-b) and leukemia-inhibitory factor (LIF), in shRNA-transduced HeLa-SFCs and found that expression of all three TGF-b isoforms was significantly downregulated while LIF remained unchanged. Expression of the Ras gene (a downstream component of TGF-b) was also markedly decreased, suggesting that the growth-inhibitory effect could be via the TGF-b pathway. The above data indicate RNA interference-based therapy may offer a new approach for CSCtargeted cancer therapy.

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“…These results are consistent with previous studies. Chang et al (37) demonstrated that siRNA-mediated suppression of HPV E6 and E7 inhibited the growth of tumor cells from cervical cancer. Sima et al (38) reported that HPV16 E7 shRNA inhibits E6 and E7 expression and induces apoptosis in cancer cells via activation of p53, p21 and Rb.…”

Section: Discussionmentioning

confidence: 99%

Induction of apoptosis by chitosan/HPV16 E7 siRNA complexes in cervical cancer cells

Yang

Guo

et al. 2012

Molecular Medicine Reports

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Abstract. Consecutive expression of the high-risk human papillomavirus (HPV) oncoproteins, E6 and E7, is pivotal for malignant transformation and maintenance of the malignant phenotype. These oncogenes may be potential targets of gene silencing-based molecular therapies for human cervical cancer. The aim of the present study was to evaluate the efficacy of chitosan-based HPV16 E7 siRNA delivery and the chitosan/HPV16 E7 siRNA complex in the induction of apoptosis in CaSki cells constitutively expressing HPV16 E6 and E7. Chitosan/siRNA nanoparticles were prepared by adding a chitosan solution drop-wise to an equal volume of siRNA solution. Formation of the chitosan/siRNA complex was verified by gel retardation assays and the entry of siRNA into the cells was confirmed by fluorescence microscopy. Expression of HPV16 E7 was examined by western blot analysis and apoptotic cells were detected by TUNEL staining. Chitosan formed complexes with HPV16 E7 siRNA. The chitosan/siRNA nanoparticles were efficiently delivered into CaSki cells and were observed to induce apoptosis. In conclusion, chitosan is suitable for use as a carrier for delivery of siRNA into cancer cells. The delivery of chitosan/HPV16 E7 siRNA nanoparticles in vivo may serve as a promising therapy for cervical cancer.

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Highly potent and specific siRNAs against E6 or E7 genes of HPV16- or HPV18-infected cervical cancers

Cited by 81 publications

References 21 publications

Human Papilloma Virus (HPV) E7-Mediated Attenuation of Retinoblastoma (Rb) Induces hPygopus2 Expression via Elf-1 in Cervical Cancer

Human Papilloma Virus (HPV) E7-Mediated Attenuation of Retinoblastoma (Rb) Induces hPygopus2 Expression via Elf-1 in Cervical Cancer

Silencing oncogene expression in cervical cancer stem-like cells inhibits their cell growth and self-renewal ability

Induction of apoptosis by chitosan/HPV16 E7 siRNA complexes in cervical cancer cells

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