Luciferase Assay to Screen Tumour-specific Promoters in Lung Cancer

Xu, Rui; Guo, Lei; Xin, Jun; Li, Wen-Mao; Gao, Yan; Zheng, You-Xian; Lin, Yujing; Xie, Ying; Wu, Yaqing; Xu, Rong

doi:10.7314/apjcp.2013.14.11.6557

Cited by 5 publications

(5 citation statements)

References 32 publications

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“…Furthermore, earlier studies have revealed an IL-4/STAT-6/Tfec/IL-4Rα positive feedback regulatory loop, in which Tfec transcribes IL-4Rα expression to promote M2 programming in macrophages, which was implicated in asthma pathogenesis ( 57 ). In addition, several genes were involved in lung cancer and other lung diseases, such as Ceacam1 ( 58 , 59 ), Ereg ( 60 , 61 ) , Selp ( 62 , 63 ), and Pik3r5 ( 64 , 65 ). The most conspicuous genes among downregulated ones are members of the keratins (KRTs) and keratin-associated proteins (KRTAPs), including Krt34 , Krtap3-1 , and Krtap7-1 , which are important for epidermal development and hair follicle morphogenesis ( 66 , 67 ), respectively.…”

Section: Resultsmentioning

confidence: 99%

Perinatal Exposure to Nicotine Alters Sperm RNA Profiles in Rats

et al. 2022

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Perinatal exposure to smoking has been associated with childhood asthma, one of the most common pediatric conditions affecting millions of children globally. Of great interest, this disease phenotype appears heritable as it can persist across multiple generations even in the absence of persistent exposure to smoking in subsequent generations. Although the molecular mechanisms underlying childhood asthma induced by perinatal exposure to smoking or nicotine remain elusive, an epigenetic mechanism has been proposed, which is supported by the data from our earlier analyses on germline DNA methylation (5mC) and histone marks (H3 and H4 acetylation). To further investigate the potential epigenetic inheritance of childhood asthma induced by perinatal nicotine exposure, we profiled both large and small RNAs in the sperm of F1 male rats. Our data revealed that perinatal exposure to nicotine leads to alterations in the profiles of sperm-borne RNAs, including mRNAs and small RNAs, and that rosiglitazone, a PPARγ agonist, can attenuate the effect of nicotine and reverse the sperm-borne RNA profiles of F1 male rats to close to placebo control levels.

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Section: Resultsmentioning

confidence: 99%

Perinatal Exposure to Nicotine Alters Sperm RNA Profiles in Rats

et al. 2022

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“…DKK1 has been reported to be a biomarker in lung cancer, with high levels of specificity and expression ( 4 ). However, our previous study demonstrated low DKK1 promoter activity in lung cancer cells ( 6 ). In the present study, 12 TREs were predicted using DHS and histone modification analysis of the lung cancer-specific DKK1 locus in A549 cells using the UCSC genome browser.…”

Section: Discussionmentioning

confidence: 85%

“…As a control, the SV40 enhancer/promoter fragment was excised from pRL-TK and inserted in forward and reverse orientations into the pGL3-basic plasmid at the Xho I and Bgl II sites to produce pGL3-SV40. Genomic DNA extraction, polymerase chain reaction (PCR) amplification and concentration quantification were performed as previously described ( 6 ). All primer sequences and cloning sites are listed in Table I .…”

Section: Methodsmentioning

confidence: 99%

“…Subcloned fragments were confirmed by sequencing. Predicted TREs were amplified by PCR according to the same protocol ( 6 ); Table II presents the primer sequences. The products were ligated into the multiple cloning site of the pGL3-DKK1-5 vector, upstream of a candidate DKK1 promoter-driven luciferase reporter gene.…”

Section: Methodsmentioning

confidence: 99%

“…The use of DKK1 and other biomarkers may increase the sensitivity of lung cancer detection. Our previous study demonstrated that the activity of the DKK1 promoter was low in lung cancer cells ( 6 ). Thus, it is likely that enhancer elements outside the basal promoter region contribute to its wide range of expression patterns ( 7 ).…”

Section: Introductionmentioning

confidence: 99%

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Prediction and identification of transcriptional regulatory elements at the lung cancer‑specific DKK1 locus

Gao

Zeng

et al. 2018

Oncol Lett

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The glycoprotein dickkopf 1 (DKK1) is highly expressed in lung cancer cell lines and tissues. Our previous study demonstrated that DKK1 promoter activity is low in lung cancer cell lines. This may be because it lacks the necessary transcriptional regulatory elements (TREs) required for higher activity levels. However, it is difficult to computationally predict functionally significant TREs, as TREs from different locations can affect large segments of distant DNA. The Encyclopedia of DNA Elements project features multiple integrated technologies and approaches for the discovery and definition of functional elements, including enhancer elements and enhancer-blocking insulators. In the present study, DNase I hypersensitive sites and histone modifications of DKK1 were investigated in the A549 lung cancer cell line using the UCSC Genome Browser. A set of cis-acting enhancer elements were identified by a dual-luciferase reporter gene assay system to increase activity of the DKK1 promoter with lung cancer specificity. To the best of our knowledge, these data provide the first insight into the role of the DKK1 locus in lung cancer, and confirm the contribution of intronic cis-acting elements to the regulation of DKK1 expression, providing a new insight into gene regulation in lung cancer, which could inform the development of targeted therapy.

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