Regulation of laryngeal squamous cell cancer progression by the lncRNA RP11‐159K7.2/miR‐206/DNMT3A axis

Wang, Xin; Yu, Boyu; Jin, Qianqian; Zhang, Junyi; Yan, Bingrui; Yang, Like; Li, Yushan; Li, Qiuying; Wang, Peng; Sun, Chuanhui; Liu, Ming; Tian, Linli; Sun, Yanan

doi:10.1111/jcmm.15331

Cited by 22 publications

(16 citation statements)

References 39 publications

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“…Total RNA from LSCC tissues and cell lines was extracted using Trizol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. The process of qRT-PCR was as previously reported [ 18 ]. Data were calculated using the 2 -ΔΔCt method.…”

Section: Methodsmentioning

confidence: 99%

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RBM15 facilitates laryngeal squamous cell carcinoma progression by regulating TMBIM6 stability through IGF2BP3 dependent

Wang

Tian

et al. 2021

J Exp Clin Cancer Res

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108

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Background Laryngeal cancer has the highest mortality rate among head and neck tumours. RNA N6-methyladenosine (m6A) is the most plentiful and variable in mammalian mRNA. Yet, the m6A regulatory mechanism underlying the carcinogenesis or progression of LSCC remains poorly understood. Methods The m6A RNA methylation quantification kit was used to detect tissue methylation levels. m6A microarray analysis, mRNA transcriptomic sequencing (mRNA-seq), and proteomics were used to determine RBM15, TMBIM6, and IGF2BP3. Immunohistochemical (IHC), quantitative real-time PCR (qRT-PCR) and Western blot were used to investigate RBM15, TMBIM6, and IGF2BP3 expression in tissue samples and cell lines. The biological effects of RBM15 were detected both in vitro and in vivo. The combination relationship between RBM15/IGF2BP3 and TMBIM6 was verified by RNA immunoprecipitation (RIP) assay, Methylated RNA immunoprecipitation sequencing (MeRIP-seq), RNase Mazf, and luciferase report assay. RNase Mazf was used to determine the methylation site on TMBIM6 mRNA. Hoechst staining assay was used to confirm the apoptotic changes. The actinomycin D verified TMBIM6 stability. Results The global mRNA m6A methylation level significantly increased in LSCC patients. RBM15, as a “writer” of methyltransferase, was significantly increased in LSCC and was associated with unfavorable prognosis. The knockdown of RBM15 reduced the proliferation, invasion, migration, and apoptosis of LSCC both in vitro and in vivo. The results were reversed after overexpressing RBM15. Mechanically, TMBIM6 acted as a downstream target of RBM15-mediated m6A modification. Furthermore, RBM15-mediated m6A modification of TMBIM6 mRNA enhanced TMBIM6 stability through IGF2BP3-dependent. Conclusion Our results revealed the essential roles of RBM15 and IGF2BP3 in m6A methylation modification in LSCC, thus identifying a novel RNA regulatory mechanism.

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Section: Methodsmentioning

confidence: 99%

“…AMC-HN-8 and TU-212 cells in 24-well plates were transfected with a luciferase reporter and indicated expression constructs. The assays were carried out as previously reported [ 18 ].…”

Section: Methodsmentioning

confidence: 99%

RBM15 facilitates laryngeal squamous cell carcinoma progression by regulating TMBIM6 stability through IGF2BP3 dependent

Wang

Tian

et al. 2021

J Exp Clin Cancer Res

Self Cite

182

108

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“…Similarly, Xin Wang et al knocked out the endogenous long noncoding RNA RP11-159K7.2 in TU-212 cells and AMC-HN-8 cells with the CRISPR-Cas9 system. The results showed that RP11-159K7.2 knockout reduced the proliferation and invasion of laryngeal squamous carcinoma cells (LSCC) in vivo and in vitro, suggesting that lncRNA RP11-159K7.2 might be a potential biomarker for LSCC therapy [91]. On the other hand, microRNAs (miRNAs) are attracting increasing attention for their role in cancer progression.…”

Section: Application Of Crispr -Cas9 In Cancer Treatmentmentioning

confidence: 99%

Review of applications of CRISPR-Cas9 gene-editing technology in cancer research

Zhao

Tong

et al. 2021

Biol Proced Online

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Characterized by multiple complex mutations, including activation by oncogenes and inhibition by tumor suppressors, cancer is one of the leading causes of death. Application of CRISPR-Cas9 gene-editing technology in cancer research has aroused great interest, promoting the exploration of the molecular mechanism of cancer progression and development of precise therapy. CRISPR-Cas9 gene-editing technology provides a solid basis for identifying driver and passenger mutations in cancer genomes, which is of great value in genetic screening and for developing cancer models and treatments. This article reviews the current applications of CRISPR-Cas9 gene-editing technology in various cancer studies, the challenges faced, and the existing solutions, highlighting the potential of this technology for cancer treatment.

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“…Luciferase reporter assay AMC-HN-8 and TU-212 cells in 24-well plates were transfected with a luciferase reporter and indicated expression constructs. The assays were carried out as previously reported [18].…”

Section: Western Blotmentioning

confidence: 99%

RBM15 facilitates laryngeal squamous cell carcinoma progression by regulating TMBIM6 stability through IGF2BP3 dependent

Wang

Tian

et al. 2021

Preprint

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Background: Laryngeal cancer has the highest mortality rate among head and neck tumours. RNA N6-methyladenosine (m6A) is the most plentiful and variable in mammalian mRNA. Yet, the m6A regulatory mechanism underlying the carcinogenesis or progression of LSCC remains poorly understood. Methods: The m6A RNA methylation quantification kit was used to detect tissue methylation levels. m6A microarray analysis, mRNA transcriptomic sequencing (mRNA-seq), and proteomics were used to determine RBM15, TMBIM6, and IGF2BP3. Immunohistochemical (IHC), quantitative real-time PCR (qRT-PCR) and Western blot were used to investigate RBM15, TMBIM6, and IGF2BP3 expression in tissue samples and cell lines. The biological effects of RBM15 were detected both in vitro and in vivo. The combination relationship between RBM15/IGF2BP3 and TMBIM6 was verified by RNA immunoprecipitation (RIP) assay, Methylated RNA immunoprecipitation sequencing (MeRIP-seq), RNase Mazf, and luciferase report assay. RNase Mazf was used to determine the methylation site on TMBIM6 mRNA. Hoechst staining assay was used to confirm the apoptotic changes. The actinomycin D verified TMBIM6 stability. Results: The global mRNA m6A methylation level significantly increased in LSCC patients. RBM15, as a “writer” of methyltransferase, was significantly increased in LSCC and was associated with unfavorable prognosis. The knockdown of RBM15 reduced the proliferation, invasion, migration, and apoptosis of LSCC both in vitro and in vivo. The results were reversed after overexpressing RBM15. Mechanically, TMBIM6 acted as a downstream target of RBM15-mediated m6A modification. Furthermore, RBM15-mediated m6A modification of TMBIM6 mRNA enhanced TMBIM6 stability through IGF2BP3-dependent. Conclusion: Our results revealed the essential roles of RBM15 and IGF2BP3 in m6A methylation modification in LSCC, thus identifying a novel RNA regulatory mechanism.

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Regulation of laryngeal squamous cell cancer progression by the lncRNA RP11‐159K7.2/miR‐206/DNMT3A axis

Cited by 22 publications

References 39 publications

RBM15 facilitates laryngeal squamous cell carcinoma progression by regulating TMBIM6 stability through IGF2BP3 dependent

RBM15 facilitates laryngeal squamous cell carcinoma progression by regulating TMBIM6 stability through IGF2BP3 dependent

Review of applications of CRISPR-Cas9 gene-editing technology in cancer research

RBM15 facilitates laryngeal squamous cell carcinoma progression by regulating TMBIM6 stability through IGF2BP3 dependent

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