Comprehensive High-Throughput RNA Sequencing Analysis Reveals Contamination of Multiple Nasopharyngeal Carcinoma Cell Lines with HeLa Cell Genomes

Strong, Michael J.; Baddoo, Melody; Nanbo, Asuka; Xu, Miao; Puetter, Adriane; Lin, Zhen

doi:10.1128/jvi.01457-14

Cited by 88 publications

(93 citation statements)

References 41 publications

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“…In this study, several considerations for utilizing HK1 include its origin from an EBV-negative NPC and p53 status, such that the cell line would be naive to the effects of EBV and mimic intact p53 mechanisms in NPC tumors (19,20,31). Although heterozygous for a mutation in p53 (codon 130), HK1 cells still can activate p53-dependent targets such as PUMA and still are responsive to p53 stabilization and growth arrest induced by Nutlin-3 treatment (20,23,(31)(32)(33) and data not shown). In stably transduced HK1 cells, LMP1 and LMP2A coexpression did not affect cell growth, with a subtle increase in motility to fibronectin (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…HK1 cells are one of the few authenticated NPC cell lines that are sensitive to serum deprivation and chemotherapeutics but can resist these cell death treatments by infection with EBV (6,23,31). Additionally, HK1 cells are free of HeLa contamination, which confounds the interpretation of many NPC-derived cell lines (32,33). To investigate cell death responses activated by DNA damage, HK1 cell lines were treated with several DNA-damaging reagents and replication stress inducers, including the topoisomerase II inhibitor etoposide, ionizing radiation, methyl methanesulfonate (MMS), and aphidicolin.…”

Section: Importancementioning

confidence: 99%

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Regulation of DNA Damage Signaling and Cell Death Responses by Epstein-Barr Virus Latent Membrane Protein 1 (LMP1) and LMP2A in Nasopharyngeal Carcinoma Cells

Wasil

Wei

Chang

et al. 2015

J Virol

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Nasopharyngeal carcinoma (NPC) is closely associated with latent Epstein-Barr virus (EBV) infection. Although EBV infection of preneoplastic epithelial cells is not immortalizing, EBVcan modulate oncogenic and cell death mechanisms. The viral latent membrane proteins 1 (LMP1) and LMP2A are consistently expressed in NPC and can cooperate in bitransgenic mice expressed from the keratin-14 promoter to enhance carcinoma development in an epithelial chemical carcinogenesis model. In this study, LMP1 and LMP2A were coexpressed in the EBV-negative NPC cell line HK1 and examined for combined effects in response to genotoxic treatments. In response to DNA damage activation, LMP1 and LMP2A coexpression reduced ␥H2AX (S139) phosphorylation and caspase cleavage induced by a lower dose (5 M) of the topoisomerase II inhibitor etoposide. Regulation of ␥H2AX occurred before the onset of caspase activation without modulation of other DNA damage signaling mediators, including ATM, Chk1, or Chk2, and additionally was suppressed by inducers of DNA single-strand breaks (SSBs) and replication stress. Despite reduced DNA damage repair signaling, LMP1-2A coexpressing cells recovered from cytotoxic doses of etoposide; however, LMP1 expression was sufficient for this effect. LMP1 and LMP2A coexpression did not enhance cell growth, with a moderate increase of cell motility to fibronectin. This study supports that LMP1 and LMP2A jointly regulate DNA repair signaling and cell death activation with no further enhancement in the growth properties of neoplastic cells. E pstein-Barr virus (EBV) is a human gammaherpesvirus that establishes lifelong latency in memory B cells, with sporadic reactivation and transmission from oral epithelia (1). More than 90% of the adult population is latently infected, and a subset can develop EBV-associated malignancies, including nasopharyngeal carcinoma (NPC), gastric cancer, Burkitt lymphoma, Hodgkin lymphoma, and lymphomas in the immunocompromised, including AIDS-associated lymphoma and posttransplant lymphoproliferative disease (2, 3). Epithelial cell infection in vitro frequently results in productive replication, and latently infected oral epithelial cells are rare in persistently infected healthy individuals (4, 5). However, epithelial tumors such as NPC consistently express a type II latency program, which includes latent membrane protein 1 (LMP1), LMP2A, and LMP2B (1, 5). Additionally, monoclonal EBV episomes are detected in NPC, suggesting that NPC tumors are the clonal outgrowth of an initially infected cell likely predisposed to oncogenic transformation from additional genetic and environmental cofactors, such as the loss of p16 and exposure to dietary nitrosamines (2, 3). In contrast to the immortalizing properties of EBV to primary B cells, the contribution of EBV infection to epithelial cell oncogenesis is less understood, as infection alone is insufficient to immortalize or induce oncogenic potential in preneoplastic cell lines from the nasopharynx (5, 6).LMP1 and LMP2A transcripts are ...

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Section: Discussionmentioning

confidence: 99%

Section: Importancementioning

confidence: 99%

Regulation of DNA Damage Signaling and Cell Death Responses by Epstein-Barr Virus Latent Membrane Protein 1 (LMP1) and LMP2A in Nasopharyngeal Carcinoma Cells

Wasil

Wei

Chang

et al. 2015

J Virol

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“…B95.8, an EBV-positive marmoset B cell line, was obtained from the American Type Culture Collection (Manassas, VA, USA) and preserved at the Cancer Research Institute, Xiangya School of Medicine (Central South University, Changsha, China) (27). CNE1 is an LMP1-negative EBV-associated epithelial carcinoma cell line that has been identified to be cross-contaminated with HeLa cells and an additional cell line of unknown origin (28). The CNE1-LMP1 cell line, which stably expresses LMP1, was obtained from the Cancer Research Institute of Central South University (26,29).…”

Section: Methodsmentioning

confidence: 99%

(-)-Epigallocatechin‑3‑gallate inhibition of Epstein‑Barr virus spontaneous lytic infection involves downregulation of latent membrane protein 1

Liu

Tang

et al. 2017

Exp Ther Med

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Abstract. The Epstein-Barr virus (EBV) lytic cycle contributes to the development of EBV-associated diseases. EBV-encoded latent membrane protein 1 (LMP1) is key to EBV lytic replication, and our previous work indicated that epigallocatechin-3-gallate (EGCG) inhibited constitutive EBV lytic infection through the suppression of LMP1-activated phosphoinositide 3-kinase/Akt and mitogen-activated protein kinase kinase/extracellular signal-related protein kinase 1/2 signaling. The present study demonstrated that LMP1 in CNE-LMP1 constructed cells significantly induced the expression of the EBV lytic proteins BZLF1 (P<0.001) and BMRF1 (P<0.05) compared with CNE1 cells. Following treatment with a specific DNAzyme that targets LMP1, significantly reduced protein expression levels of BZLF1 and BMRF1 in EBV-associated epithelial carcinoma CNE1-LMP1 cells (P<0.001 and P<0.01, respectively) and lymphoma B95.8 cells (both P<0.01) were observed. Furthermore, EGCG significantly inhibited the mRNA and protein expression levels of LMP1 (P<0.05) in an apparent dose-dependent manner in CNE1-LMP1 and B95.8 cells. Thus, the present findings indicated that the molecular mechanism underlying EGCG inhibition of EBV lytic infection involves downregulation of LMP1. IntroductionEpstein-Barr virus (EBV) is a human herpes virus that infects over 90% of the human population worldwide (1). EBV is suggested to be an environmental factor associated with the development of several human malignancies, including nasopharyngeal carcinoma (NPC), Burkitt's lymphoma, Hodgkin's disease (HD), gastric cancer, natural killer/T-cell lymphoma, acquired immune deficiency syndrome-and transplantation-associated lymphoma (2,3), breast carcinoma, and hepatocellular carcinoma (4,5). EBV, as all other herpes viruses, may establish a latent or lytic infection in host cells (6). Notably, studies suggest that EBV reactivation into a lytic cycle may contribute to the pathogenesis of malignancies (7,8). EBV lytic infection in vivo has been identified by elevated antibody titers against EBV lytic antigens and by increased viral DNA load in the serum/plasma, and these observations correspond with advanced cancer stages, poor prognosis or tumor recurrence following therapy (9,10). Additionally, serological studies have indicated that EBV lytic infection may occur months or years prior to a clinical diagnosis of NPC, HD or Burkitt's lymphoma, which suggests EBV lytic infection may be a risk factor for cancer development (11-13).Green tea contains (-)-epigallocatechin-3-gallate (EGCG), which is reported to have antioxidant, antibacterial and antitumor effects (14,15). EGCG has been indicated to modulate multiple signaling pathways, including the phosphoinositide 3-kinase (PI3-K)/Akt and mitogen-activated protein kinase (MAPK) signaling pathways, which may enable it to exert its cancer chemopreventive and therapeutic effects (16,17).EBV encoded latent membrane protein 1 (LMP1), which is considered to have oncogenic properties, has been identified in 90% of patients with NPC...

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“…We selected 14 paired samples that exhibited higher increase in expression of HOTAIR in tumor over their paired normal tissues. We analyzed HOTAIR transcripts by transcript per million (TPM) and percentage of each isoform in the tumor samples using RSEM (Li and Dewey, 2011; Strong et al ., 2014). In nine tumor samples, HOTAIR‐N exhibited the highest expression among all three isoforms (Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…The datasets were then analyzed using the RSEM algorithm for the quantification of isoforms of HOTAIR transcripts as previously described (Li and Dewey, 2011; Strong et al ., 2014). …”

Section: Methodsmentioning

confidence: 99%

Induction of a novel isoform of the lncRNA HOTAIR in Claudin‐low breast cancer cells attached to extracellular matrix

Zhuang

et al. 2017

Molecular Oncology

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Elevated overexpression of the lncRNA HOTAIR mediates invasion and metastasis in breast cancer. In an apparent paradox, we observed low expression of HOTAIR in the invasive Claudin‐low MDA‐MB‐231 and Hs578T cells in two‐dimensional culture (2D). However, HOTAIR expression exhibited robust induction in laminin‐rich extracellular matrix‐based three‐dimensional organotypic culture (lrECM 3D) over that in 2D culture. Induction of HOTAIR required intact ECM signaling, namely integrin α2 and SRC kinase activity. Moreover, invasive growth was suppressed by HOTAIR‐specific siRNA. Induction of HOTAIR in lrECM 3D culture resulted from the activation of a novel isoform of HOTAIR (HOTAIR‐N) whose transcription is started from the first intron of the HOXC11 gene. The HOTAIR‐N promoter exhibited increased trimethylation of histone H3 lysine 4, a histone marker of active transcription, and binding of BRD4, a reader of transcriptionally active histone markers. Inhibition of BRD4 substantially reduced the expression of HOTAIR in lrECM 3D culture. In summary, our results indicate that HOTAIR expression is activated by BRD4 binding to a novel HOTAIR‐N promoter in Claudin‐low breast cancer cells that are attached to ECM. Induction of HOTAIR is required for invasive growth of Claudin‐low breast cancer cells in lrECM 3D culture.

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Comprehensive High-Throughput RNA Sequencing Analysis Reveals Contamination of Multiple Nasopharyngeal Carcinoma Cell Lines with HeLa Cell Genomes

Cited by 88 publications

References 41 publications

Regulation of DNA Damage Signaling and Cell Death Responses by Epstein-Barr Virus Latent Membrane Protein 1 (LMP1) and LMP2A in Nasopharyngeal Carcinoma Cells

Regulation of DNA Damage Signaling and Cell Death Responses by Epstein-Barr Virus Latent Membrane Protein 1 (LMP1) and LMP2A in Nasopharyngeal Carcinoma Cells

(-)-Epigallocatechin‑3‑gallate inhibition of Epstein‑Barr virus spontaneous lytic infection involves downregulation of latent membrane protein 1

Induction of a novel isoform of the lncRNA HOTAIR in Claudin‐low breast cancer cells attached to extracellular matrix

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