Analyses of P16INK4a gene promoter methylation relative to molecular, demographic and clinical parameters characteristics in non-small cell lung cancer patients: A pilot study

Jurišić, Vladimir; Obradovic, Jasmina; Nikolic, Nadja; Javorac, Jovan; Perin, Branislav; Milašin, Jelena

doi:10.1007/s11033-022-07982-1

Cited by 7 publications

(4 citation statements)

References 41 publications

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“…Furthermore, the promoter hypermethylation of the p16 gene leads to gene silencing, which is of great significance in confirming the downregulation of p16 protein expression in NSCLC ( 18 ). However, more than half of NSCLC failed to be detected with p16 Cgi methylation, indicating that the inactivation of p16 in lung cancer might be only partially caused by abnormal DNA methylation that occurs in the p16 Cgi region ( 19 ). Actually, methylation of Cgi shore has been shown to be inversely associated with gene expression, similar to the function of Cgi ( 20 ), However, few studies have focused on the changes of DNA methylation in regions outside the Cgi of p16 .…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, the significant role of p16 in the development and progression of LUAD cannot be solely attributed to these minor changes in methylation levels. Therefore, it is difficult to differentiate between LUAD and normal tissues solely based on p16 Cgi methylation, which is a commonly used method for detecting cancer in humans ( 19 ). To gain a deeper understanding, it is essential to investigate which specific region of p16 is crucial for DNA methylation to exert its function, as along with exploring the underlying mechanism involved.…”

Section: Discussionmentioning

confidence: 99%

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The transcription activity of OTX2 on p16 expression is significantly blocked by methylation of CpG shore in non-promoter of lung cancer cell lines

Peng,

Fu,

Chang

et al. 2023

Transl Cancer Res

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Background The aberrant expression of the classical tumor suppressor gene p16 is a frequent event in lung cancer mainly due to the hypermethylation of its 5’-cytosine-phosphate-guanine-3’ island (Cgi). However, whether methylation happens in other regions and how p16 expression and function are affected are largely unknown. Methods Clustered Regularly Interspaced Short Palindromic Repeats/dCas9 (CRISPR/dCas9) technology was used for methylation editing at specific site of p16 . The effects of methylation editing were detected by 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium, inner salt (MTS), transwell migration and wound healing tests. Chromatin immnoprecipitation-quantitative polymerase chain reaction (CHIP-qPCR) was performed to explore the impact of Cgi shore methylation on the binding abilities of transcription factors (TFs) including YY1 , SP1 , ZNF148 and OTX2 to p16 gene. A rescue experiment was performed to verify the regulatory effect of OTX2 on p16 . The negative relationship between p16 expression and the methylation level of Cgi shore in non-promoter region was further verified with datasets from The Cancer Genome Atlas (TCGA) program and lung adenocarcinoma (LUAD) patients’ samples. Results The suppressive effect of p16 Cgi shore methylation on its expression was demonstrated in both HEK293 and A549 cells using CRISPR/dCas9-mediated specific site methylation editing. Methylation of the Cgi shore in the p16 non-promoter region significantly decreased its expression and promoted cell growth and migration. The ability of OTX2 bound to p16 was significantly reduced by 19.35% after methylation modification. Over-expression of OTX2 in A549 cells partly reversed the inhibitory effect of methylation on p16 expression by 19.04%. The verification results with TCGA and LUAD patients’ samples supported that the p16 Cgi shore is a key methylation regulatory region. Conclusions Our findings suggested that methylation of the Cgi shore in the p16 non-promoter region can hamper the transcriptional activity of OTX2 , leading to a reduction in the expression of p16 , which might contribute to the development of lung cancer.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

The transcription activity of OTX2 on p16 expression is significantly blocked by methylation of CpG shore in non-promoter of lung cancer cell lines

Peng,

Fu,

Chang

et al. 2023

Transl Cancer Res

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“…Non-small cell lung cancer (NSCLC), including adenocarcinoma, squamous cell carcinoma and large cell carcinoma, comprises ~85% of all lung cancer cases (1,2). Advances in recent decades have led to the identification of numerous oncogenic factors in NSCLC, such as gene mutations in EGFR, BRAF, ROS proto-oncogene 1, p16 INK4a and human epidermal growth factor receptor 2, and rearrangements in anaplastic lymphoma kinase and RET (3)(4)(5)(6) Although target therapies and immunotherapies have enhanced the overall survival time of patients, the occurrence of metastasis and resistance (primary or acquired) may cause difficulties for effective second-line treatment options. This has resulted in an urgent unmet need for NSCLC treatment (7).…”

Section: Introductionmentioning

confidence: 99%

Doxazosin inhibits vasculogenic mimicry in human non‑small cell lung cancer through inhibition of the VEGF‑A/VE‑cadherin/mTOR/MMP pathway

Hsu,

Leu,

Hsu

et al. 2024

Oncol Lett

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Lung cancer is the leading cause of cancer-related death worldwide, and ~85% of lung cancers are non-small cell lung cancer (NSCLC), which has a low 5-year overall survival rate and high mortality. Several therapeutic strategies have been developed, such as targeted therapy, immuno-oncotherapy and combination therapy. However, the low survival rate indicates the urgent need for new NSCLC treatments. Vasculogenic mimicry (VM) is an endothelial cell-free tumor blood supply system of aggressive and metastatic tumor cells present during tumor neovascularization. VM is clinically responsible for tumor metastasis and resistance, and is correlated with poor prognosis in NSCLC, making it a potential therapeutic target. In the present study, A549 cells formed glycoprotein-rich lined tubular structures, and transcript levels of VM-related genes were markedly upregulated in VM-forming cells. Based on a drug repurposing strategy, it was demonstrated that doxazosin (an antihypertensive drug) displayed inhibitory activity on VM formation at non-cytotoxic concentrations. Doxazosin significantly reduced the levels of vascular endothelial growth factor A (VEGF-A) and matrix metalloproteinase-2 (MMP-2) in the cell media during VM formation. Further experiments revealed that the protein expression levels of VEGF-A and vascular endothelial-cadherin (VE-cadherin), which contribute to tumor aggressiveness and VM formation, were downregulated following doxazosin treatment. Moreover, the downstream signaling Ephrin type-A receptor 2 (EphA2)/AKT/mTOR/MMP/Laminin-5γ2 network was inhibited in response to doxazosin treatment. In conclusion, the present study demonstrated that doxazosin displayed anti-VM activity in an NSCLC cell model through the downregulation of VEGF-A and VE-cadherin levels, and the suppression of signaling pathways related to the receptor tyrosine kinase, EphA2, protein kinases, AKT and mTOR, and proteases, MMP-2 and MMP-9. These results support the add-on anti-VM effect of doxazosin as a potential agent against NSCLC.

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“… 15 , 16 In some countries, smoking is widespread and is associated with lung tumors. 17 , 18 Recently, cigarette smoking has not only be regarded as a predisposing factor for the occurrence of HCC, but it has also been investigated as an independent predictor of mortality and been shown to increase the risk of development of HCC in people with HCV or HBV infections and individuals with alcoholism. 19 - 21 However, the association between cigarette smoking and recurrence after curative resection in HCC has not yet been well investigated and is still controversial.…”

Section: Introductionmentioning

confidence: 99%

Smoking as a Risk Factor for Very Late Recurrence in Surgically Resected Early-Stage Primary Hepatocellular Carcinoma

Cho,

Wang,

Tsai

et al. 2024

Clin Med Insights Oncol

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Background: The risk of first recurrence of hepatocellular carcinoma (HCC) within years 5 to 10 after curative hepatectomy remains unknown. We aimed to assess the incidence and prognostic factors for very late recurrence among patients who achieved 5 years’ recurrence-free survival (RFS) after primary resection. Methods: We retrospectively analyzed 337 patients with early-stage HCC underwent primary tumor resection and achieved more than 5 years’ RFS. Results: A total of 77 patients (22.8%) developed very late recurrence. The cumulative very late recurrence rate increased from 6.9% and 11.7% to 16.6% at 6, 7, and 8 years, respectively. Patients stopped smoking had a higher rate of very late RFS. Conclusions: The high rates of very late recurrence in HCC indicate that patients warrant continued surveillance, even after 5 recurrence-free years. Moreover, smoking is a risk factor for very late HCC recurrence, and quitting smoking may reduce the risk of very late recurrence.

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Analyses of P16INK4a gene promoter methylation relative to molecular, demographic and clinical parameters characteristics in non-small cell lung cancer patients: A pilot study

Cited by 7 publications

References 41 publications

The transcription activity of OTX2 on p16 expression is significantly blocked by methylation of CpG shore in non-promoter of lung cancer cell lines

The transcription activity of OTX2 on p16 expression is significantly blocked by methylation of CpG shore in non-promoter of lung cancer cell lines

Doxazosin inhibits vasculogenic mimicry in human non‑small cell lung cancer through inhibition of the VEGF‑A/VE‑cadherin/mTOR/MMP pathway

Smoking as a Risk Factor for Very Late Recurrence in Surgically Resected Early-Stage Primary Hepatocellular Carcinoma

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