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Special RCA based sensitive point-of-care detection of HPV mRNA for cervical cancer screening
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作者 Yi Long Shurui Tao +5 位作者 Dongni Shi Xingyu Jiang Tian Yu Yingxi Long libing song Guozhen Liu 《Aggregate》 EI CAS 2024年第4期399-415,共17页
Developing the sensitive point-of-care testing(POCT)of oncogenic nucleic acids from human papillomavirus(HPV)infection is essential in preventing cervical can-cer,especially in resource-limited settings.Rolling circle... Developing the sensitive point-of-care testing(POCT)of oncogenic nucleic acids from human papillomavirus(HPV)infection is essential in preventing cervical can-cer,especially in resource-limited settings.Rolling circle amplification(RCA)is attractive in achieving POCT via nucleic acid-based aggregation under isothermal conditions.However,the influence of RCA product structure on the aggregation remains unexplored resulting in limited sensitivity.Here,a minimum secondary structured RCA technique(MSS-RCA)is developed by designing a unique circu-lar template,demonstrating significantly enhanced detection sensitivity with only one amplification step and one primer under isothermal conditions.The amplifi-cation efficiency of MSS-RCA could be kinetically manipulated by controlling the secondary structure of the circular template.Introducing the invertase probe to MSS-RCA,HPV16 E6/E7 nucleic acid target was detected with a personal glucose meter(PGM)with a sensitivity of 5 fM(50 zmol in 10µL).This integrated MSS-RCA-PGM detection system was successfully applied to detect HPV16 E6/E7 mRNA extracted from 54 cervical swab samples reaching a positive predictive value of 100.00%and negative predictive values of 96.00%(77.77%to 99.40%,95%CI).MSS-RCA-PGM provides a sensitive POCT platform for the detection of nucleic acid biomarkers for screening of cervical cancer or other diseases. 展开更多
关键词 cervical cancer screening HPV mRNA nucleic acid-based aggregation point-of-care detection rolling circle amplification sensitivity
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Up-regulation of miR-1245 by c-myc targets BRCA2 and impairs DNA repair
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作者 libing song Ting Dai +7 位作者 Yingjun Xie Chanjuan Wang Chuyong Lin Zhiqiang Wu Zhe Ying Jueheng Wu Mengfeng Li Jun Li 《Journal of Molecular Cell Biology》 SCIE CAS CSCD 2012年第2期108-117,共10页
BRCA2,a clinical prognostic factor,is significantly up-regulated in mRNA level,while its protein expression is often decreased in sporadic breast cancer.However,how BRCA2 protein expressions are suppressed in these tu... BRCA2,a clinical prognostic factor,is significantly up-regulated in mRNA level,while its protein expression is often decreased in sporadic breast cancer.However,how BRCA2 protein expressions are suppressed in these tumors remains unknown.In this study,we demonstrated that miR-1245 directly suppressed BRCA23′-UTR and translation,impaired homologous recombination(HR)-mediated repair,reduced DNA damage-induced Rad51 nuclear foci,and rendered cells hypersensitive to g-irradiation(IR),ul-timately inducing high chromosomal abnormalities in normal breast cells and breast cancer cells.Conversely,inhibiting miR-1245 in breast cancer cells enhanced BRCA2 levels and induced resistance to IR.Furthermore,we demonstrated that c-myc up-regulated miR-1245 expression via direct binding to the miR-1245 promoter,which led to down-regulation of BRCA2 and reduction in HR efficiency.Significantly,miR-1245 levels in primary breast tumors correlated with c-myc overexpression and BRCA2 suppression.These findings uncover a BRCA2 regulatory and signaling pathway in sporadic breast cancer and support a functionally and clinically relevant epigenetic mechanism in cancer pathogenesis. 展开更多
关键词 BRCA2 miR-1245 DNA damage C-MYC
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