Expression of RAR β2 gene by real-time RT-PCR: Differential expression in normal subjects compared to cervical cancer patients normalised against GAPDH as a housekeeping gene

Abu, Jafaru; Batuwangala, Madu; Symonds, Paul

doi:10.1016/j.ejogrb.2008.04.009

Cited by 3 publications

(2 citation statements)

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“…In conclusion, we demonstrate that under certain in vitro cellular backgrounds and in the cervix of young K14E7 transgenic mice, the HPV16 E7 oncoprotein induces the RARB gene, as previously observed in benign cervical lesions [38]. The mechanism involved in RARB overexpression induced by the HPV16 E7 oncoprotein needs further investigation.…”

Section: Discussionmentioning

confidence: 66%

“…For example, in CC development, the RARB transcription decrease is correlated with methylation increase of its promoter, from 11 % in low-grade to 29 % in high-grade lesions and up to 33-63 % in invasive cancers [23,24]. In this regard, one study reported that transcription of this important gene shows a 1.5-fold increase in benign cervical lesions, while a progressive reduction of RARB expression was seen with advanced stage of the disease [38]. On the other hand, in foreskin keratinocytes immortalized with HPV16 or HPV18, there is a sequential RARB promoter methylation and a decrease at the RARB mRNA level, although this methylation did not become manifested until late passages of these cells [39].…”

Section: Discussionmentioning

confidence: 97%

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Human papillomavirus type 16 E7 oncoprotein upregulates the retinoic acid receptor-beta expression in cervical cancer cell lines and K14E7 transgenic mice

et al. 2015

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Persistent infection with high-risk human papillomaviruses is the main etiological factor in cervical cancer (CC). The human papillomavirus type 16 (HPV16) E7 oncoprotein alters several cellular processes, regulating the expression of many genes in order to avoid cell cycle control. Retinoic acid receptor beta (RARB) blocks cell growth, inducing differentiation and apoptosis. This tumor suppressor gene is gradually silenced in late passages of foreskin keratinocytes immortalized with HPV16 and in various tumors, including CC, mainly by epigenetic modifications. We investigated the effect of E7 oncoprotein on RARB gene expression. We found that HPV16 E7 increases RARB mRNA and RAR-beta protein expression both in vitro and in the cervix of young K14E7 transgenic mice. In E7-expressing cells, RARB overexpression is further increased in the presence of the tumor suppressor p53 (TP53) R273C mutant. This effect does not change when either C33-A or E7-expressing C33-A cell line is treated with Trichostatin A, suggesting that E7 enhances RARB expression independently of histone deacetylases inhibition. These findings indicate that RARB overexpression is part of the early molecular events induced by the E7 oncoprotein.

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Section: Discussionmentioning

confidence: 66%

Section: Discussionmentioning

confidence: 97%

Human papillomavirus type 16 E7 oncoprotein upregulates the retinoic acid receptor-beta expression in cervical cancer cell lines and K14E7 transgenic mice

et al. 2015

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Bioreactor environment‐sensitive sentinel genes as novel metrics for cell culture scale‐down comparability

Kondragunta

Joshi

Han

et al. 2012

Biotechnology Progress

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Scale-down of bioreactors is currently done based on matching one or more measurable parameters such as k(L) a and P/V, which could result in insufficient process comparability. Currently, there is a lack of genomic translational studies in cell culture scale-down, which could help delineate measurable cellular attributes for improved scale-down. In this study, we scaled-down from a typical bench-scale 5-L bioreactor to a novel high-throughput 35-mL minibioreactor based on matching oxygen transfer rate, which resulted in cell growth and product-related discrepancies using Sp2/0 cells. Performing DNA microarrays on time-course samples from both systems, we identified ∼200 differentially expressed transcripts, presumably because of bioreactor aeration and mixing differences with scale-down. Evaluating these transcripts for bioreactor-relevant cellular functions such as oxidative stress response and DNA damage response, we chose 18 sentinel genes based on their degree of difference and functionality, which we further verified by quantitative real-time polymerase chain reaction (qRT-PCR). Tracking the differential expression of Sod1, Apex1, and Odc1 genes, we were able to correlate sparging-related damage and poor mixing, as possible causes for physiological changes such as prolonged culture in minibioreactors. Additionally, to verify our sentinel gene findings, we performed follow-up improved scale-down studies based on gene analysis and measured transcriptomic changes. As a result, qRT-PCR-based genomic profiles and cell growth profiles showed better convergence between the improved minibioreactor conditions and the model 5-L bioreactor. Our results broadly show that based on the knowledge from transcriptomic changes of sentinel gene profiles, it is possible to improve bioreactor scale-down for more comparable processes.

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CIP2A expression is elevated in cervical cancer

Huang

Adelson

Mordechai

et al. 2011

CBM

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Early detection of cervical cancer is critical for a favorable prognosis. Standard cytological detection methods, such as Pap smear, are highly subjective and HPV detection is not a reliable marker for predicting the malignancy potential of cervical lesions. As a result, there is a demand for a diagnostic assay capable of sensitive and specific detection of cervical cancer. In this preclinical exploratory study, qRT-PCR and western blotting were used to assess expression levels of CIP2A and p16INK4a in cervical tissue samples (n(normal adjacent) = 23, n(tumor) = 29). CIP2A was abundantly expressed in cervical cancer cell lines and was not expressed in normal epithelial cells. CIP2A mRNA levels were higher in cervical tumor tissues in comparison to the level of CIP2A mRNA in normal adjacent tissue from cervical cancer patients. CIP2A protein was specifically expressed in cervical tumor tissues at different cancer grades and stages, and was not observed in normal adjacent tissue. Elevated CIP2A mRNA levels in cervical tissues had a sensitivity of 80% and specificity of 91% and CIP2A protein expression detection had a sensitivity of 83% and specificity of 100%, similar to that of p16INK4a, with no correlation of CIP2A expression with HPV infection, age, race, or other patient characteristics. However the number of samples analyzed in this preliminary study is limited and a large prospective cohort study is necessary to further evaluate CIP2A as a biomarker for cervical cancer.

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Expression of RAR β2 gene by real-time RT-PCR: Differential expression in normal subjects compared to cervical cancer patients normalised against GAPDH as a housekeeping gene

Cited by 3 publications

References 4 publications

Human papillomavirus type 16 E7 oncoprotein upregulates the retinoic acid receptor-beta expression in cervical cancer cell lines and K14E7 transgenic mice

Human papillomavirus type 16 E7 oncoprotein upregulates the retinoic acid receptor-beta expression in cervical cancer cell lines and K14E7 transgenic mice

Bioreactor environment‐sensitive sentinel genes as novel metrics for cell culture scale‐down comparability

CIP2A expression is elevated in cervical cancer

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