Aberrant promoter methylation reduced the expression of protocadherin 17 in nasopharyngeal cancer

2019 ◽  
Vol 97 (4) ◽  
pp. 364-368
Author(s):  
Shiming Yang ◽  
Zhirao Dai ◽  
Weimin Li ◽  
Rongguan Wang ◽  
Dongyan Huang
Keyword(s):  
Flow Cytometry ◽  
Cell Lines ◽  
Nasopharyngeal Cancer ◽  
Underlying Mechanism ◽  
Tissue Samples ◽  
Adjacent Tissue ◽  
Normal Epithelial Cell ◽  
Proliferation And Apoptosis ◽  
Epithelial Cell Lines ◽  
Pcr Method

This study aimed to explore the underlying mechanism of protocadherin 17 (PCDH17) downregulated in nasopharyngeal cancer (NPC). NPC tumor and adjacent tissue samples were collected from NPC patients who received therapy in the Chinese PLA General Hospital. Meanwhile, a normal epithelial cell lines NP96 and 6 NPC and cell lines C666-1, CEN1, CEN2, HNE1, HEN2, and HONE1 were prepared. Then, the expression level of PCDH17 and the methylation level of PCDH17 promoter in both tissues samples and cell lines were determined using the PCR method. Moreover, PCDH17 was overexpressed in CNE2 and HONE1 using Lipo2000. Following this, the proliferation and apoptosis of CNE2 and HONE1 were assessed using MTT and flow cytometry. The expression of PCDH17 was significantly downregulated in NPC tissues compared with the adjacent tissues as well as in the NPC cell lines compared with the normal NP96 cells. Overexpressed PCDH17 could significantly inhibit the proliferation of CNE2 and HONE1 cells but obviously promote the apoptosis of these two cell lines. Aberrant hypermethylation in the promoter might be the explanation of PCDH17 downregulated in PCDH17 and promoted the development of NPC.

scholarly journals Circular RNA hsa_circ_0032463 acts as a tumor promoter in osteosarcoma by regulating miR-498/LEF1 axis

2021 ◽  
Author(s):  
Guanghua Qin ◽  
Xuejian Wu
Keyword(s):  
Wound Healing ◽  
Cell Lines ◽  
Underlying Mechanism ◽  
Circular Rna ◽  
Inhibitory Effect ◽  
Experimental Results ◽  
Tumor Promoter ◽  
Qpcr Analysis ◽  
Proliferation And Apoptosis ◽  
The Relationship

Several studies have examined the relationship between osteosarcoma (OS) and microRNAs (miRNAs). However, only a few researchers have investigated the underlying mechanism of circRNAs in OS development. Our paper aimed to assess how circ_0032463 initiates and regulates OS progression. We detected circ_0032463 expression in OS tissues and cell lines using RT-qPCR analysis and then investigated the interaction between circ_0032463, miR-498 and LEF1 using RNA pull-down, RIP, and luciferase assays. The effect of the circ_0032463/miR-498/LEF1 axis on the migration, proliferation, and apoptosis levels of OS cells was explored using CCK-8, BrdU, wound healing, and FITC assays. Our findings revealed that circ_0032463 expression was upregulated in OS tissues and cell lines. We also found that circ_0032463 interacted with miR-498, thereby reducing the expression of miR-498 in OS cells. Experimental results indicated that miR-498 could directly target LEF1 in OS cells and that circ_0032463 could abrogate the tumor-inhibitory effect of miR-498 by upregulating LEF1 in OS. More specifically, by binding to miR-498 and inhibiting LEF1 expression, circ_0032463 promoted the migration and proliferation abilities of OS cells and suppressed the apoptosis ability of OS cells. Overall, this research suggested that circ_0032463 could promote OS development by regulating the miR-498/LEF1 axis.

scholarly journals Assessment of Global Methylation in Paraffin Embedded Prostatic Tissues and Cell Lines Using Flow Cytometry

2018 ◽  
Vol 1 (1) ◽  
pp. 34-38
Author(s):  
Osama Sharaf Eldin ◽  
Mahmoud M. Elfar ◽  
Abdel-Motaal Fouda
Keyword(s):  
Flow Cytometry ◽  
Cell Lines ◽  
Objective Assessment ◽  
Prostate Adenocarcinoma ◽  
Cell Nuclei ◽  
Global Methylation ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Global Hypomethylation ◽  
Prostatic Cell

Background. The aim of this study is to measure global 5-methylcystosine (5MeC) methylation in paraffin embedded prostatic tissues and cell lines using flow cytometry. Methods. Cell/nuclei suspension from 10 cases of benign prostatic hyperplasia (BPH), 10 cases of prostatic adenocarcinoma, and two prostatic cell lines (PNT1A and LNCaP) were prepared using modified heat pretreatment technique. 5MeC global methylation was assessed by flow cytometry of cell/nuclei suspension and immunostaining of tissue sections. Results. Higher percentage of positively stained cells (PPSC) and mean channel fluorescence (MCF) were detected in PNT1A cell line and BPH cell/nuclei suspensions as compared to LNCaP cell lines and adenocarcinoma cell/nuclei suspensions. Lower scores of 5MeC immunostaining were observed in all prostate adenocarcinoma tissue sections as compared to BPH sections indicating global hypomethylation in prostate adenocarcinoma. Two distinctive populations of cells were detected in histograms generated from most of the BPH cell/nuclei suspensions. Conclusion. The study developed a novel technique that could measure 5MeC global methylation in paraffin embedded prostatic tissues. This represents a rapid and objective assessment of methylation and when combined with tissue micro-dissection and cell sorting, this technique could be applied to larger tissue samples such as post radical prostatectomy and transurethral resected specimens.

scholarly journals Epigenetic reprogramming underlies efficacy of DNA demethylation therapy in osteosarcomas

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Naofumi Asano ◽  
Hideyuki Takeshima ◽  
Satoshi Yamashita ◽  
Hironori Takamatsu ◽  
Naoko Hattori ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cpg Islands ◽  
Underlying Mechanism ◽  
Dna Demethylation ◽  
Epigenetic Reprogramming ◽  
Epigenetic Therapy ◽  
Tissue Samples ◽  
Multiple Tumor ◽  
Microarray Screening

AbstractOsteosarcoma (OS) patients with metastasis or recurrent tumors still suffer from poor prognosis. Studies have indicated the efficacy of DNA demethylation therapy for OS, but the underlying mechanism is still unclear. Here, we aimed to clarify the mechanism of how epigenetic therapy has therapeutic efficacy in OS. Treatment of four OS cell lines with a DNA demethylating agent, 5-aza-2′-deoxycytidine (5-aza-dC) treatment, markedly suppressed their growth, and in vivo efficacy was further confirmed using two OS xenografts. Genome-wide DNA methylation analysis showed that 10 of 28 primary OS had large numbers of methylated CpG islands while the remaining 18 OS did not, clustering together with normal tissue samples and Ewing sarcoma samples. Among the genes aberrantly methylated in primary OS, genes involved in skeletal system morphogenesis were present. Searching for methylation-silenced genes by expression microarray screening of two OS cell lines after 5-aza-dC treatment revealed that multiple tumor-suppressor and osteo/chondrogenesis-related genes were re-activated by 5-aza-dC treatment of OS cells. Simultaneous activation of multiple genes related to osteogenesis and cell proliferation, namely epigenetic reprogramming, was considered to underlie the efficacy of DNA demethylation therapy in OS.

scholarly journals High impact of miRNA-4521 on FOXM1 expression in medulloblastoma

2019 ◽  
Vol 10 (10) ◽  
Author(s):  
Daniel Senfter ◽  
Mahzeiar Samadaei ◽  
Robert M. Mader ◽  
Johannes Gojo ◽  
Andreas Peyrl ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Cell Lines ◽  
Cell Cycle Progression ◽  
Cellular Proliferation ◽  
Underlying Mechanism ◽  
Cyclin B1 ◽  
Tissue Samples ◽  
Medulloblastoma Cell ◽  
Foxm1 Expression ◽  
Medulloblastoma Cell Lines

Abstract Medulloblastoma, an embryonal tumor of the cerebellum/fourth ventricle, is one of the most frequent malignant brain tumors in children. Although genetic variants are increasingly used in treatment stratification, survival of high-risk patients, characterized by leptomeningeal dissemination, TP53 mutation or MYC amplification, is still poor. FOXM1, a proliferation-specific oncogenic transcription factor, is deregulated in various solid tumors, including medulloblastoma, and triggers cellular proliferation, migration and genomic instability. In tissue samples obtained from medulloblastoma patients, the significant upregulation of FOXM1 was associated with a loss of its putative regulating microRNA, miR-4521. To understand the underlying mechanism, we investigated the effect of miR-4521 on the expression of the transcription factor FOXM1 in medulloblastoma cell lines. Transfection of this microRNA reduced proliferation and invasion of several medulloblastoma cell lines and induced programmed cell death through activation of caspase 3/7. Further, downstream targets of FOXM1 such as PLK1 and cyclin B1 were significantly reduced thus affecting the cell cycle progression in medulloblastoma cell lines. In conclusion, a restoration of miRNA-4521 may selectively suppress the pathophysiological effect of aberrant FOXM1 expression and serve as a targeted approach for medulloblastoma therapy.

scholarly journals MALAT1 promotes gastric adenocarcinoma through the MALAT1/miR-181a-5p/AKT3 axis

Open Biology ◽  
2019 ◽  
Vol 9 (9) ◽  
pp. 190095 ◽  
Author(s):  
Zhengmao Lu ◽  
Tianhang Luo ◽  
Tao Pang ◽  
Zongxin Du ◽  
Xiaoyi Yin ◽  
...  
Keyword(s):  
Gastric Adenocarcinoma ◽  
Cell Lines ◽  
Protein Level ◽  
Underlying Mechanism ◽  
Clonogenic Survival ◽  
Specific Protein ◽  
Malignant Tumours ◽  
Non Coding Rna ◽  
Proliferation And Apoptosis ◽  
Long Non Coding Rna

Gastric adenocarcinoma, which originates from the gastric mucosal epithelium, has the highest incidence among various malignant tumours in China. As a crucial long non-coding RNA, metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) has been suggested to play an important role in many tumours. Here, we aimed to investigate the role and underlying mechanism of MALAT1 in gastric adenocarcinoma. Quantitative reverse transcription polymerase chain reaction was applied to determine the expression levels of MALAT1 in serum and cell lines. A CCK-8 assay and a clonogenic survival assay were used to examine cell proliferation and apoptosis. The protein level of RAC-γ serine/threonine-specific protein kinase (AKT3) was determined by western blot. Our results showed that MALAT1 was highly expressed in the serum of patients with gastric adenocarcinoma and in cell lines. Downregulating MALAT1 inhibited proliferation and promoted apoptosis of MGC-803 cells. In addition, MALAT1 directly targeted and decreased the expression of miR-181a-5p, which in turn upregulated the expression of AKT3. Further, overexpressing miR-181a-5p or directly inhibiting the AKT pathway with the inhibitor ipatasertib exhibited similar effects to MALAT1 knockdown. Our research proposes a novel mechanism where the role of MALAT1 is dependent on the MALAT1/miR-181a-5p/AKT3 axis. MALAT1 competes with AKT3 for miR-181a-5p binding, thereby upregulating the AKT3 protein level and ultimately promoting the growth of gastric adenocarcinoma.

scholarly journals Establishment of normal epithelial cell lines from mouse submandibular gland in serum-free cell culture.

1990 ◽  
Vol 36 (4) ◽  
pp. 791-801
Author(s):  
Masafumi YABUMOTO ◽  
Tetsuji OKAMOTO ◽  
Teruhiko OSAKI ◽  
Yoshinari MYOKEN ◽  
Kazuaki TAKADA
Keyword(s):  
Cell Culture ◽  
Epithelial Cell ◽  
Submandibular Gland ◽  
Cell Lines ◽  
Free Cell ◽  
Serum Free ◽  
Normal Epithelial Cell ◽  
Epithelial Cell Lines
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