Role of Mutation of NOTCH1 gene in development of oral squamous cell carcinoma: a narrative review.

Cancer of the oral cavity has numerous types and, among all, oral squamous cell carcinoma represents >90% of all cancers of the oral area. Oral squamous cell carcinoma arises from the squamous lining of the oral cavity. Across the globe, most commonly it develops in the regions of tongue followed by floor of the mouth, and lower lip. Neurogenic locus notch homolog protein 1 gene has its association with oral squamous cell carcinoma and is known to be associated with both oncogenic and tumour suppressor roles. The current narrative review comprised literature published from 2013 to 2023. It was searched on Google Scholar, PubMed and Google databases. Globally, neurogenic locus notch homolog protein 1 mutations are associated with the development of oral squamous cell carcinoma. Most of the mutations are linked to ligand bind epidermal growth factor-like repeat region of extracellular domain of neurogenic locus notch homolog protein 1. Once activated, the pathway is involved in tumour progression and metastasis. The Asians compared to Caucasians are more affected by neurogenic locus notch homolog protein 1 mutations.

Relevance research between the expression of p16INK4a , Notch1, and hTERC genes: The development of HPV16-positive cervical cancer.

BACKGROUND: GLOBOCAN 2018 latest data show cervical cancer ranks fourth in morbidity and mortality among women. Many genes in cervical lesions differ in sensitivity and specificity. However, the diagnostic molecules for early cervical cancer are not very clear. This paper screens biomarkers for early molecular diagnosis of Mongolian patients with cervical cancer. METHODS: Immunohistochemical SP method was used to detect the expression of p16INK4a and Notch1 protein in paraffin sections of 226 Mongolian patients with HPV16-positive cervical lesions after pathological examination, and 100 of them were randomly selected by fluorescence in situ hybridization to detect hTERC gene. The HPV16-binding human cervical cancer SiHa cell line was used to silence the expression of HPV16 E6/E7 gene by RNA interference, and the expression of p16INK4a , Notch1, and hTERC genes and protein expression levels were detected by RT-PCR and Western blot. RESULTS: The positive expression rates of p16INK4a , Notch1, and hTERC genes in HPV16-positive cervical cancer, CIN-III, CIN-II, CIN-I, uterine leiomyoma, and chronic cervicitis were significantly different (P < .05); the positive expression rates of the three genes were also significantly different in the same type of cervical lesions (P < .05); RNA interference can effectively inhibit HPV16 E6/E7, p16INK4a and Notch1 gene expression, but has no effect on hTERC gene expression. CONCLUSION: The p16INK4a gene can be used as a biomarker for early screening of cervical cancer, and the hTERC gene can be used to confirm the clinical diagnosis of cervical cancer.

Sequencing of NOTCH1 gene in an Italian population with bicuspid aortic valve: Preliminary results from the GISSI OUTLIERS VAR study.

BACKGROUND: Bicuspid aortic valve (BAV) formation is genetically determined, with reduced penetrance and variable expressivity. NOTCH1 is a proven candidate gene and its mutations have been found in familial and sporadic cases of BAV. METHODS: 66 BAV patients from the GISSI VAR study were genotyped for the NOTCH1 gene. RESULTS: We identified 63 variants, in heterozygous and homozygous states. Fifty-two are common polymorphisms present in almost all patients. Eleven variants are new and never yet reported: two are non-synonymous substitutions, Gly540Asp in exon 10 and Glu851Gln in exon 16; one is in the 3'UTR region and seven in introns, one corresponds to a T allele insertion in intron 27. We selected four statistically noteworthy and seven new variants identified in six BAV patients and correlated them with clinical and demographic variables and with imaging and histological parameters. Preliminary data show that four were BAV patients with isolated stenosis in patients over 60 aged. These variants may correlate with a later need for surgery for the presence of stenosis and not aortic valve regurgitation or ascending aortic aneurysm. CONCLUSIONS: Completing the genotyping of 62 BAV patients we found 11 new variants in the NOTCH1 gene never yet reported. These findings confirm that the identification of new, clinically remarkable biomarkers for BAV requires a deeper genetic understanding of the NOTCH1 gene variants, which could be targeted by future diagnostic and therapeutic strategies.

Familial aggregation of "apple peel" intestinal atresia and cardiac left-sided obstructive lesions: A possible causal relationship with NOTCH1 gene mutations.

"Apple peel" intestinal atresia is a rare form of small bowel atresia, in which the duodenum or proximal jejunum ends in a blind pouch and the distal small bowel wraps around its vascular supply, in a spiral resembling an apple peel. The etiology of "apple peel" intestinal atresia is presently unknown, although a congenital or acquired intestinal vascular accident can have a role in the pathogenesis. We report a family in which the proband affected by "apple peel" intestinal atresia, had a sibling (an interrupted pregnancy), and a paternal cousin with cardiac left-sided obstructive lesions. Molecular testing for NOTCH1 gene was carried out in the proband, because pathogenic mutations in this gene have been associated with familial and sporadic cardiac left-sided obstructive lesions and vascular anomalies, both isolated or within the spectrum of the Adams-Oliver syndrome (AOS). The heterozygous c.2734C>T (p.Arg912Trp) NOTCH1 variant was found in the proband with "apple peel" intestinal atresia and in his father. This result argues for a possible causal relationship between NOTCH1 gene mutations and some forms of intestinal defects, through a vascular mechanism. The spectrum of NOTCH1-associated malformations is widened. Genetic counseling should take into account intrafamilial variable clinical expression and incomplete penetrance.

Mutaciones del gen NOTCH1 en la leucemia linfocítica crónica / NOTCH1 gene mutations in chronic lymphocytic leukemia

Chronic lymphocytic leukemia (CLL) is the most common leukemia in the Western world. The disease has\r\na highly variable clinical course, ranging from very indolent cases to patients with aggressive and rapidly\r\nprogressing outcome. Genetic studies are useful tools in analyzing this pathology, and have been incorporated in international risk classifications. The analysis of genomic rearrangements and the mutational status of immunoglobulin heavy chain variable have allowed risk groups of high prognostic value to be established. More recently, next generation sequencing studies have identified novel somatic mutations that could explain the wide clinical variability of this pathology. Among them, the analysis of NOTCH1 (neurogenic locus notch homolog protein 1) gene mutations are of interest, as deregulation is associated with tumorigenesis. NOTCH11 mutations are mostly located at exon 34 (80% of cases) and 3´UTR (untranslated region). They produce premature stop codons that produce a constitutively active and stable NOTCH1 protein. NOTCH1 mutations are associated with adverse prognosis and refractoriness to treatment. The aim of this study was to analyze NOTCH1 mutations in CLL patients by ASO-PCR and sequencing. Our results found 4.4% of cases with NOTCH1 mutated values concordant with international observations (5%-10%). Including them in the genetic status of CLL patients allows the characterization of risk groups, an aspect of great importance in clinical practice and therapeutic decisions, to be refined.

La leucemia linfocítica crónica (LLC) es la leucemia más frecuente en adultos de Occidente. Presenta\r\nun curso clínico altamente variable, con pacientes que requieren tratamiento inmediato y otros con un curso indolente de la enfermedad. Los estudios genéticos constituyen herramientas de suma utilidad en esta enfermedad, encontrándose incorporados a las clasificaciones de riesgo internacionales. El análisis de los rearreglos genómicos y del estado mutacional de los genes IGHV (immunoglobulin heavy chain variable region) ha hecho factible establecer grupos de riesgo de alto valor pronóstico. Más recientemente, estudios de secuenciación de última generación permitieron la detección de mutaciones\r\nsomáticas previamente desconocidas en esta afección, que podrían explicar la amplia variabilidad clínica\r\nobservada en la LLC. Entre ellas, resultan de interés las observadas en el gen NOTCH1 (neurogenic locus notch homolog protein 1), cuya desregulación se asocia con el desarrollo tumoral. Estas mutaciones se acumulan en mayor medida en el exón 34 (80% de los casos) y en la región 3´UTR (untraslated region), lo que genera codones de terminación prematuros que originan una proteína NOTCH1 constitutivamente activa y más estable, los cuales se asocian con pronóstico adverso y refractariedad al tratamiento. Nuestro objetivo fue evaluar mutaciones de NOTCH1 en nuestros pacientes mediante ASO-PCR y secuenciación. Se detectaron mutaciones en el 4.4% de los casos, valor concordante con los datos internacionales (5% a 10%). Su inclusión en la caracterización genética de los pacientes con LLC permitirá refinar la categorización de los grupos de riesgo, aspecto de suma importancia tanto en el seguimiento clínico como en la toma de decisiones terapéuticas.

Notch1 targeted regulation of mir-224/LRIG2 signaling for the proliferation and apoptosis of cervical cancer cells.

In the present study, we explored the participation of Notch1 targeted regulation of mir-224/LRIG2 gene signal pathway in proliferation and apoptosis of cervical cancer cells. Forty-nine cases of cervical cancer lesion samples from cervical cancer patients treated in our hospital from February 2013 to February 2015 were chosen as subjects (the observation group), and cervical samples of healthy women (42 cases) during the same period were used as the control group. We determined the mRNA and protein expression of Notch1, mir-224, and LRIG2 genes. We also analyzed the mutual relationship between Notch1 gene expression and cervical cancer. The Notch1 genes in the cervical cancer cells (HeLa) were silenced and overexpressed to measure cancer apoptosis with flow cytometry. After obstruction of the Notch1 signal pathway, the mRNA and protein expression in the mir-224 and LRIG2 genes was also measured. It was found that in comparison to the control group, Notch1 gene expression in the observation group was significantly higher (p<0.05), cell growth was suppressed in Notch1 silent cell strains but accelerated in overexpressed Notch1 cells. The silencing of Notch1 genes can lead to the reduction of mir-224/LRIG gene and protein levels, while overexpression of the Notch1 genes increased the mir-224/LRIG gene and protein levels. In conclusion, the Notch1 gene is positively related to cervical cancer and can promote the occurrence of the disease. The potential mechanism shows that Notch1 gene can regulate cervical cancer cell proliferation by regulating the mir-224/LRIG2 signal pathway.

Targeted inhibition of Notch1 gene enhances the killing effects of paclitaxel on triple negative breast cancer cells.

OBJECTIVE: To study the influence of targeted inhibition of Notch1 gene on the killing effects of paclitaxel on triple negative breast cancer cells. METHODS: The triple negative [estrogen receptor (ER)/progesterone receptor (PR)/human epidermal growth factor receptor 2 (Her2)] breast cancer cell line MDA-MB-231 and ER/PR/HER-2-positive breast cancer cell line MCF-7 were cultured, transfected with Notch1-siRNA-overexpression plasmid and blank plasmid, and treated with different concentrations of paclitaxel, and then the cell proliferation activity and apoptosis rate as well as the mRNA expression of Caspase-3, Caspase-9 and Bcl-2 were determined. RESULTS: Paclitaxel could decrease the MDA-MB-231 and MCF-7 cell proliferation activity as well as Bcl-2 mRNA expression, and increase MDA-MB-231 and MCF-7 cell apoptosis rate as well as Caspase-3 and Caspase-9 mRNA expression in dose-dependent manners; with the same dose of paclitaxel treatment, the inhibitory effects on MDA-MB-231 cell proliferation activity and Bcl-2 mRNA expression as well as the promoting effects on MDA-MB-231 cell apoptosis and mRNA expression of Caspase-3 and Caspase-9 were weaker than those on MCF-7 cell; after 0.5 µM paclitaxel combined with Notch1-siRNA treatment, MDA-MB-231 cell proliferation activity and Bcl-2 mRNA expression were significantly lower than those after 0.5 µM paclitaxel combined with control plasmid treatment while cell apoptosis rate and mRNA expression of Caspase-3 and Caspase-9 were higher than those after 0.5 µM paclitaxel combined with control plasmid treatment. CONCLUSIONS: Targeted inhibition of Notch1 gene may enhance the killing effects of paclitaxel on triple negative breast cancer cells by up-regulating the expression of Caspase-3 and Caspase-9 and inhibiting the expression of Bcl-2.

Associations between polymorphisms in the NICD domain of bovine NOTCH1 gene and growth traits in Chinese Qinchuan cattle.

NOTCH1 is one of the four mammalian Notch receptors, which is involved in the Notch signaling pathway. Specifically, NOTCH1 promotes the proliferation of myogenic precursor cells, and the NICD domain of NOTCH1 can impair regeneration of skeletal muscles. However, similar research on the bovine NOTCH1 gene is lacking. In this study, we detected the polymorphisms of the bovine NOTCH1 gene in a total of 448 individuals from Chinese Qinchuan cattle with DNA pooling, forced PCR-RFLP, and DNA sequencing methods. Five novel SNPs were identified within the NICD domain, and eight haplotypes comprising combinations of these five SNPs were studied as well. The association analysis of SNPs' effects with growth traits revealed that g.A48250G was significantly associated with body height, body weight, and height at hip cross, and that g.A49239C only showed significant associations with body height. This suggests that the NOTCH1 gene is a strong candidate gene that could be utilized as a promising marker in beef cattle breeding programs.

Analysis of variants located in 3'UTR regions of NOTCH1 and JAG1 genes for children with conotruncal heart defects / 上海交通大学学报（医学版）

Objective · To explore the correlation between variants located in 3' untranslated regions (3'UTR) of NOTCH1 and JAG1 genes and conotruncal heart defects (CTD). Methods · Six hundred CTD children without 22q11 deletion and three hundred healthy children were enrolled in this hospital-based case-control study. Variants located in the 3'UTR regions of NOTCH1 and JAG1 genes were detected by high-throughput sequencing. The accuracy of the variants were verified by PCR and Sanger sequencing. Online software PicTar, TargetScan and microRNA.org were used to make functional predictions. Results · One mutation and three SNPs were found in the 3'UTR of NOTCH1. Three mutations and six SNPs were found in the 3'UTR of JAG1. The genotypic distributions of two SNPs (rs3840074 and rs8708) located in JAG13'UTR between CTD group and the controls were statistically significant (both P<0.05). Results of prediction showed that all the four mutations and two meaningful SNPs could bind to microRNA. Conclusion · The variants located in 3'UTR regions of NOTCH1 and JAG1 genes may be related to the occurrence of CTD.

Effect of silencing Notch1 gene on proliferation, apoptosis and Akt/mTOR pathyway inhibition in clear cell renal cell carcinoma / 中华泌尿外科杂志

Objective To investigate the effect of silencing Notch1 gene by RNA interference on the proliferation,apoptosis and Akt/mTOR signaling pathway in clear renal cell carcinoma.Methods The optimal segment targeting Notch1 gene was designed and transfected into 786-O cells by Lipofectamine TM2000.The Notch1 mRNA and protein were detected by RT-PCR and Western blot.The proliferation rate of 786-O cells was evaluated by MTT and the variation of apoptosis was measured by TUNEL.The protein expression level of apoptosis-related protein Bcl-2,caspase-3,caspase-9,and signaling pathway protein Akt,p-Akt,p-mTOR,p-P70S6K were detected by Western blott.Results Notchl mRNA and protein was markedly suppressed by the siRNA targeting Notch1.Treated with 0,40,60,80,100 and 120 nmol/L of Notch1 siRNA for 24 hours,cell proliferation rates were (98.51 ± 1.33) ％,(87.34 ± 2.26) ％,(64.72 ± 3.24)％,(57.68 ±3.32)％,(31.91 ± 1.85)％ and (19.27 ±2.73)％,and the difference was statistically significant (P ＜ 0.01).Treated with 0,40,80,and 120 nmol/L of Notchl siRNA for 24 hours,apoptosis rates were (7.6 ± 3.8) ％,(21.5 ± 4.8) ％,(32.3 ± 3.5) ％,and (46.3 ± 4.7％),the difference was statistically significant (P ＜ 0.05).Decreased expression of Akt signaling pathway proteins p-Akt,p-mTOR,p-70S6K and apoptosis-related protein Bcl-2,procaspase-3 was detected,but no change in the total protein of Akt.Conclusions Depletion of Notch1 gene could inhibit cell growth and induce apoptosis in 786-O cell line.It inhibits Akt/mTOR signaling pathway by dephosphorylated.

Targeted inhibition of Notch1 gene enhances the killing effects of paclitaxel on triple negative breast cancer cells

OBJECTIVE@#To study the influence of targeted inhibition of Notch1 gene on the killing effects of paclitaxel on triple negative breast cancer cells.@*METHODS@#The triple negative [estrogen receptor (ER)/progesterone receptor (PR)/human epidermal growth factor receptor 2 (Her2)] breast cancer cell line MDA-MB-231 and ER/PR/HER-2-positive breast cancer cell line MCF-7 were cultured, transfected with Notch1-siRNA-overexpression plasmid and blank plasmid, and treated with different concentrations of paclitaxel, and then the cell proliferation activity and apoptosis rate as well as the mRNA expression of Caspase-3, Caspase-9 and Bcl-2 were determined.@*RESULTS@#Paclitaxel could decrease the MDA-MB-231 and MCF-7 cell proliferation activity as well as Bcl-2 mRNA expression, and increase MDA-MB-231 and MCF-7 cell apoptosis rate as well as Caspase-3 and Caspase-9 mRNA expression in dose-dependent manners; with the same dose of paclitaxel treatment, the inhibitory effects on MDA-MB-231 cell proliferation activity and Bcl-2 mRNA expression as well as the promoting effects on MDA-MB-231 cell apoptosis and mRNA expression of Caspase-3 and Caspase-9 were weaker than those on MCF-7 cell; after 0.5 μM paclitaxel combined with Notch1-siRNA treatment, MDA-MB-231 cell proliferation activity and Bcl-2 mRNA expression were significantly lower than those after 0.5 μM paclitaxel combined with control plasmid treatment while cell apoptosis rate and mRNA expression of Caspase-3 and Caspase-9 were higher than those after 0.5 μM paclitaxel combined with control plasmid treatment.@*CONCLUSIONS@#Targeted inhibition of Notch1 gene may enhance the killing effects of paclitaxel on triple negative breast cancer cells by up-regulating the expression of Caspase-3 and Caspase-9 and inhibiting the expression of Bcl-2.

Targeted inhibition of Notch1 gene enhances the killing effects of paclitaxel on triple negative breast cancer cells

Objective To study the influence of targeted inhibition of Notch1 gene on the killing effects of paclitaxel on triple negative breast cancer cells. Methods The triple negative [estrogen receptor (ER)/progesterone receptor (PR)/human epidermal growth factor receptor 2 (Her2)] breast cancer cell line MDA-MB-231 and ER/PR/HER-2-positive breast cancer cell line MCF-7 were cultured, transfected with Notch1-siRNA-overexpression plasmid and blank plasmid, and treated with different concentrations of paclitaxel, and then the cell proliferation activity and apoptosis rate as well as the mRNA expression of Caspase-3, Caspase-9 and Bcl-2 were determined. Results Paclitaxel could decrease the MDA-MB-231 and MCF-7 cell proliferation activity as well as Bcl-2 mRNA expression, and increase MDA-MB-231 and MCF-7 cell apoptosis rate as well as Caspase-3 and Caspase-9 mRNA expression in dose-dependent manners; with the same dose of paclitaxel treatment, the inhibitory effects on MDA-MB-231 cell proliferation activity and Bcl-2 mRNA expression as well as the promoting effects on MDA-MB-231 cell apoptosis and mRNA expression of Caspase-3 and Caspase-9 were weaker than those on MCF-7 cell; after 0.5 μM paclitaxel combined with Notch1-siRNA treatment, MDA-MB-231 cell proliferation activity and Bcl-2 mRNA expression were significantly lower than those after 0.5 μM paclitaxel combined with control plasmid treatment while cell apoptosis rate and mRNA expression of Caspase-3 and Caspase-9 were higher than those after 0.5 μM paclitaxel combined with control plasmid treatment. Conclusions Targeted inhibition of Notch1 gene may enhance the killing effects of paclitaxel on triple negative breast cancer cells by up-regulating the expression of Caspase-3 and Caspase-9 and inhibiting the expression of Bcl-2.

Effect of Notch1 gene knockdown on migration and proliferation of human prostate cancer cell line PC-3 / 肿瘤研究与临床

Objective To investigate the effect of Notch1 gene dowuregulated on migration and proliferation of human prostate cancer cell line PC-3.Methods The small interfering RNA (siRNA) targeted Notch1 gene or negative control sequences was transfected into PC-3 cells.The expression of Notch1 or Hes1 gene was detected by Real-Time PCR and Western Blot.Then ability of migration or proliferation was measured by Transwell assay or MTS Assay.Results Compared with negative control group (36.097±1.941) and untransfected group (38.762±1.897),Notch1 expression level in siRNA group (3.960±0.510) was significantly reduced (P ＜ 0.01).Meanwhile,Hes1 level in siRNA group was decreased,expression in three groups as follows:siRNA group was 1.690±0.994,negative control group was 8.776±0.916,untransfected group was 9.803±1.001 (P ＜ 0.01).In Transwell assay,the number of migration cells in siRNA group was 657.867±27.610,more than that in the negative control group (158.533±18.263) and untransfected group (146.933±15.733) (P ＜ 0.01).In MTS assay,there was no significant difference among three groups at 0 h point,however,siRNA group was significantly raised at the time points of 24,48 and 72 h (P ＜ 0.01).Conclusions Downregulation of Notch1 gene by transfection of the siRNA-Notch1 sequences significantly promoted ability of migration or proliferation in PC-3 cells,and the effect may be due to the down-regulation of Hes1 expression.

TGF-ß1 induces a nucleus pulposus-like phenotype in Notch 1 knockdown rabbit bone marrow mesenchymal stem cells.

We have investigated the effects of Notch1 knockdown and treatment with TGF-ß1 on the regulation of the directional differentiation of mesenchymal stem cells (MSCs). MSCs were isolated from the femur bone of New Zealand rabbits and purified by using discontinuous gradient density centrifugation. Notch1 siRNAs were designed, synthesised and transiently transfected into these MSCs, and treated with TGF-ß1. The MSCs were examined for morphology, stained with toluidine blue for proteoglycan analysis and gene and protein expression were measured with qRT-PCR and Western blotting respectively. They had an ellipse or fusiform shape and gathered in nests or swirls after being cultured for 7 days. Notch1 expression was knocked down with Notch1 siRNA (the silence rate was 47%; P < 0.001). After knockdown and TGF-ß1 treatment, MSCs expressed more proteoglycan (P < 0.01), and higher levels of collagen II mRNA and protein than control cells (P < 0.001). Thus knockdown of Notch1 expression in MSCs may be useful in the treatment of intervertebral disc degeneration.

Mutation of Notch1 gene and its protein expression in T-cell lymphoma / 白血病·淋巴瘤

Objective To investigate the effect of the expression of Notch1 protein and the mutation of Notch1 gene in.T-cell lymphoma (TCL).Methods Immunohistochemistry was used to detect the expression of Notch1 protein,and PCR amplification and DNA sequencing were used to detect the mutation of Notch1 gene in the 26th and 27th HD domain and the 34th PEST domain in 30 cases.10 cases of reactive hyperplasia tissues of lymph node were as the control.Results The positive rates of Notch1 protein expression and Notch1 gene mutation were 70.0 ％ (21/30) and 56.7 ％ (17/30).8 cases of Notch1 mutations were detected in the HD domain,6 cases in the PEST domain,and 3 case in both HD and PEST domains.Inscrtion,deletion,nonsense mutation and missense mutation were included in Notch 1 mutations.Conclusion Notch1 gene mutation may play an important role in the expression of Notch1 protein.The occur of TCL is related to the expression of Notch1 protein and the mutation of Notch1 gene.

Role of Notch - 1 in the Differentiation of Rat Mesenchymal Stem Cells into Neurons / 重庆医科大学学报

Objectives; To identify the roles of Notch - 1 gene for survivaland differentiation of mesenchymal stem cells ( MSCs) into neurons by way of RNA interference ( RNAi) technique to suppress expression of Notch - 1 gene. Methods; Firstly, construct mNotch - 1 short hairpin RNA (mNotch -IshRNA) and then to oberve configuration of MSCs transfected by mNotch - IshRNA; MTT was employed to evaluate the survival ratio of MSCs before transfection, 24 hours and 3 days after transfection, respectively; agents, such as BDNF, retinoid acid, were obtained to induce MSCs transfected by mNotch - IshRNA in vitro and expression of Nestin, NSE and NF200( neuron marker agent) and GFAP( neural glial cell marker agent) were determined by immunocytochemistry stain technique. Results: The cell cubic configuration was enforced and expression of notch - 1 gene vanished by transfection of mNotch -IshRNA. The survival ratio of MSCs by transfection decreased and there was significant differences between non - ransfection and transfection - controlling group after 24 hours and 3 days of transfection ( P