scholarly journals MPS1 is involved in the HPV16-E7-mediated centrosomes amplification

Cell Division ◽  
2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Yair Alfaro-Mora ◽  
Guadalupe Domínguez-Gómez ◽  
Rodrigo E. Cáceres-Gutiérrez ◽  
Laura Tolentino-García ◽  
Luis A. Herrera ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Western Blot ◽  
Cell Line ◽  
Protein Stabilization ◽  
The Other ◽  
Protein Levels ◽  
Human Papillomavirus Type 16 ◽  
Qrt Pcr ◽  
Transcriptional Target

Abstract Background It has been reported that the oncoprotein E7 from human papillomavirus type 16 (HPV16-E7) can induce the excessive synthesis of centrosomes through the increase in the expression of PLK4, which is a transcriptional target of E2F1. On the other hand, it has been reported that increasing MPS1 protein stability can also generate an excessive synthesis of centrosomes. In this work, we analyzed the possible role of MPS1 in the amplification of centrosomes mediated by HPV16-E7. Results Employing qRT-PCR, Western Blot, and Immunofluorescence techniques, we found that E7 induces an increase in the MPS1 transcript and protein levels in the U2OS cell line, as well as protein stabilization. Besides, we observed that inhibiting the expression of MPS1 in E7 protein-expressing cells leads to a significant reduction in the number of centrosomes. Conclusions These results indicate that the presence of the MPS1 protein is necessary for E7 protein to increase the number of centrosomes, and possible implications are discussed.

scholarly journals Loss of HOXB3 promotes development of hormone receptor negative breast cancer

2020 ◽  
Author(s):  
Lizhe Zhu ◽  
Shibo Yu ◽  
Siyuan Jiang ◽  
Guanqun Ge ◽  
Yu Yan ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Western Blot ◽  
Cell Line ◽  
Hormone Receptor ◽  
Poor Prognosis ◽  
Cancer Gene ◽  
Qrt Pcr ◽  
Lower Expression ◽  
Breast Cancer Gene

Abstract BackgroundThe homobox (HOX) gene family as a transcription factor encoding a specific nuclear protein is essential for embryonic development, differentiation, and homeostasis. The role of HOXB3 protein varies in different tumors. This study aims to explore the role of the HOXB3 gene in breast cancer.MethodDifferentiated expressed genes were screened by analyzing metastatic breast cancer gene chip data in TCGA and GEO database. The function of selected HOXB3 gene was also analyzed by GEPIA, Kaplan-Meier Plotter, Breast Cancer Gene-Expression Miner and metascape. Molecular biology methods such as qRT-PCR, western blot and IF was used to verify bio-informatics findings.ResultsBoth bio-informatics analyses and western blot showed that HOXB3 was lost in breast cancer compared to normal breast tissue. Survival analysis also showed that lower expression of HOXB3 was associated with poor prognosis. Bio-informatics analyses further showed that HOXB3 was positively correlated with hormone receptors. qRT-PCR, immunofluorescence and western blot also confirmed that HOXB3 had the highest expression in the immortalized breast epithelial cell line MCF-10A, lower in luminal breast cancer cell line T47D and the lowest in triple negative breast cancer (TNBC) cell line MDA-MB-231. Metascape for GO analysis of GEO data provided possible mechanism that HOXB3 could positively regulate cell adhesion, inhibit cell proliferation and activate immune response in breast cancer, and considered that HOXB3 might cause cell malignant transformation through the above pathways.ConclusionIn summary, HOXB3 expression was decreased in breast cancer, especially in hormone receptor-negative breast cancer. The lower expression of HOXB3 was associated with poor prognosis. It might become a new biomarker to predict prognosis of breast cancer.

scholarly journals Knockdown of lncRNA X inactive specific transcript (XIST) radiosensitizes non-small cell lung cancer (NSCLC) cells through regulation of miR-16-5p/WEE1 G2 checkpoint kinase (WEE1) axis

2021 ◽  
Vol 35 ◽  
pp. 205873842096608
Author(s):  
Ran Du ◽  
Feng Jiang ◽  
Yanhua Yin ◽  
Jinfen Xu ◽  
Xia Li ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Western Blot ◽  
Small Cell ◽  
Luciferase Reporter ◽  
Small Cell Lung ◽  
Luciferase Reporter Gene ◽  
G2 Checkpoint ◽  
Qrt Pcr ◽  
Specific Transcript

Long non-coding RNA (lncRNA) X inactive specific transcript (XIST) is reported to play an oncogenic role in non-small cell lung cancer (NSCLC). However, the role of XIST in regulating the radiosensitivity of NSCLC cells remains unclear. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expressions of XIST and miR-16-5p in NSCLC in tissues and cells, and Western blot was used to assess the expression of WEE1 G2 checkpoint kinase (WEE1). Cell counting kit-8 (CCK-8), colony formation and flow cytometry assays were used to determine cell viability and apoptosis after NSCLC cells were exposed to different doses of X-rays. The interaction between XIST and miR-16-5p was confirmed by StarBase database, qRT-PCR and dual-luciferase reporter gene assays. TargetScan database was used to predict WEE1 as a target of miR-16-5p, and their targeting relationship was further validated by Western blot, qRT-PCR and dual-luciferase reporter gene assays. XIST was highly expressed in both NSCLC tissue and cell lines, and knockdown of XIST repressed NSCLC cell viability and cell survival, and facilitated apoptosis under the irradiation. MiR-16-5p was a target of XIST, and rescue experiments demonstrated that miR-16-5p inhibitors could reverse the role of XIST knockdown on radiosensitivity in NSCLC cells. WEE1 was validated as a target gene of miR-16-5p, and WEE1 could be negatively regulated by XIST. XIST promotes the radioresistance of NSCLC cells by regulating the expressions of miR-16-5p and WEE1, which can be a novel target for NSCLC therapy.

scholarly journals E1∧E4 Protein of Human Papillomavirus Type 16 Associates with Mitochondria

2004 ◽  
Vol 78 (13) ◽  
pp. 7199-7207 ◽  
Author(s):  
Kenneth Raj ◽  
Samuel Berguerand ◽  
Shirley Southern ◽  
John Doorbar ◽  
Peter Beard
Keyword(s):  
Human Papillomavirus ◽  
Terminal Portion ◽  
Hpv Dna ◽  
Human Papillomavirus Type 16 ◽  
Dead Box ◽  
Virion Production ◽  
Severe Reduction ◽  
Initial Binding ◽  
Binding To Mitochondria

ABSTRACT The human papillomavirus (HPV) E1∧E4 protein is the most abundantly expressed viral protein in HPV-infected epithelia. It possesses diverse activities, including the ability to bind to the cytokeratin network and to DEAD-box proteins, and in some cases induces the collapse of the former. E1∧E4 is also able to prevent the progression of cells into mitosis by arresting them in the G2 phase of the cell cycle. In spite of these intriguing properties, the role of this protein in the life cycle of the virus is not clear. Here we report that after binding to and collapsing the cytokeratin network, the HPV type 16 E1∧E4 protein binds to mitochondria. When cytokeratin is not present in the cell, E1∧E4 appears associated with mitochondria soon after its synthesis. The leucine cluster within the N-terminal portion of the E1∧E4 protein is pivotal in mediating this association. After the initial binding to mitochondria, the E1∧E4 protein induces the detachment of mitochondria from microtubules, causing the organelles to form a single large cluster adjacent to the nucleus. This is followed by a severe reduction in the mitochondrial membrane potential and an induction of apoptosis. HPV DNA replication and virion production occur in terminally differentiating cells which are keratin-rich, rigid squamae that exfoliate after completion of the differentiation process. Perturbation of the cytokeratin network and the eventual induction of apoptotic properties are processes that could render these unyielding cells more fragile and ease the exit of newly synthesized HPVs for subsequent rounds of infection.

scholarly journals Knockdown of Long Non-Coding RNA XIST Inhibited Doxorubicin Resistance in Colorectal Cancer by Upregulation of miR-124 and Downregulation of SGK1

2018 ◽  
Vol 51 (1) ◽  
pp. 113-128 ◽  
Author(s):  
Jia Zhu ◽  
Rui Zhang ◽  
Dongxiang Yang ◽  
Jibin Li ◽  
Xiaofei Yan ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Western Blot ◽  
Molecular Mechanisms ◽  
Luciferase Reporter ◽  
Regulatory Function ◽  
Glutathione S Transferase ◽  
Protein Levels ◽  
Qrt Pcr ◽  
Ic50 Value

Background/Aims: Doxorubicin (DOX) is a widely used chemotherapeutic agent for colorectal cancer (CRC). However, the acquirement of DOX resistance limits its clinical application for cancer therapy. Mounting evidence has suggested that aberrantly expressed lncRNAs contribute to drug resistance of various tumors. Our study aimed to explore the role and molecular mechanisms of lncRNA X-inactive specific transcript (XIST) in chemoresistance of CRC to DOX. Methods: The expressions of XIST, miR-124, serum and glucocorticoid-inducible kinase 1 (SGK1) mRNA in DOX-resistant CRC tissues and cells were detected by qRT-PCR or western blot analysis. DOX sensitivity was assessed by detecting IC50 value of DOX, the protein levels of P-glycoprotein (P-gp) and glutathione S-transferase-π (GST-π) and apoptosis. The interactions between XIST, miR-124 and SGK1 were confirmed by luciferase reporter assay, qRT-PCR and western blot. Xenograft tumor assay was used to verify the role of XIST in DOX resistance in CRC in vivo. Results: XIST expression was upregulated and miR-124 expression was downregulated in DOX-resistant CRC tissues and cells. Knockdown of XIST inhibited DOX resistance of CRC cells, as evidenced by the reduced IC50 value of DOX, decreased P-gp and GST-π levels and enhanced apoptosis in XIST-silenced DOX-resistant CRC cells. Additionally, XIST positively regulated SGK1 expression by interacting with miR-124 in DOX-resistant CRC cells. miR-124 suppression strikingly reversed XIST-knockdown-mediated repression on DOX resistance in DOX-resistant CRC cells. Moreover, SGK1-depletion-elicited decrease of DOX resistance was greatly restored by XIST overexpression or miR-124 inhibition in DOX-resistant CRC cells. Furthermore, XIST knockdown enhanced the anti-tumor effect of DOX in CRC in vivo. Conclusion: XIST exerted regulatory function in resistance of DOX possibly through miR-124/SGK1 axis, shedding new light on developing promising therapeutic strategy to overcome chemoresistance in CRC patients.

Cloning and sequencing of the murine farnesyltransferase α-encoding cDNA from a cell line which expresses the human papillomavirus type-16 E6 gene

Gene ◽  
1995 ◽  
Vol 164 (2) ◽  
pp. 373-374 ◽  
Author(s):  
Hiroshi Shirasawa ◽  
Tomoaki Kinoshita ◽  
Yuji Shino ◽  
Kohji Mori ◽  
Kumiko Shimizu ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Cell Line ◽  
Cloning And Sequencing ◽  
Human Papillomavirus Type 16 ◽  
E6 Gene ◽  
A Cell

The interaction between steroid hormones, human papillomavirus type 16, E6 oncogene expression, and cervical cancer

2003 ◽  
Vol 13 (6) ◽  
pp. 834-842 ◽  
Author(s):  
M. Moodley ◽  
S. Sewart ◽  
C. S. Herrington ◽  
R. Chetty ◽  
R. Pegoraro ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Steroid Hormones ◽  
Housekeeping Gene ◽  
Hpv 16 ◽  
Tissue Samples ◽  
Hpv Dna ◽  
Human Papillomavirus Type 16 ◽  
Transforming Proteins

Various risk factors have been implicated in the causation of cervical cancer including human papillomavirus (HPV), the early genes (E6 and E7) of which encode the main transforming proteins. Studies have suggested that steroid hormones may enhance the expression of these genes leading to loss of p53 gene-mediated cell apoptosis. A total of 120 cervical tissue samples were obtained from patients with proven cervical cancer. Patients who used depo-medroxyprogesterone acetate steroid contraception were recruited as part of the steroid arm. Only HPV DNA type 16 samples were used for the study. Controls included three cell lines (CaSki, SiHa, & C33A) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal housekeeping gene. Of 120 patients, there were 111 patients with HPV type 16 identified. Of this number, RNA was present in 63 samples. There were 30 women (30/63) who used steroid contraception. In relation to patients who used contraception, HPV 16 E6 gene expression was present in 79% (n = 23) and 88% (n = 30) of steroid users compared to nonusers, respectively. In total there were 25 patients (40%) with expression of the HPV 16 E6*I gene and 30 patients with expression of the E6*II gene. There were 57% of steroid users (n = 17) who had expression of the E6*I/E6*II gene, compared to 52% (n = 17) of nonusers (P = 0.800). From a molecular level, this study does not confirm the role of injectable progesterones in cervical carcinogenesis.

scholarly journals Role of Calpain in the Formation of Human Papillomavirus Type 16 E1^E4 Amyloid Fibers and Reorganization of the Keratin Network

2011 ◽  
Vol 85 (19) ◽  
pp. 9984-9997 ◽  
Author(s):  
J. Khan ◽  
C. E. Davy ◽  
P. B. McIntosh ◽  
D. J. Jackson ◽  
S. Hinz ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Amyloid Fibers ◽  
Human Papillomavirus Type 16

scholarly journals Establishment of the Human Papillomavirus Type 16 (HPV-16) Life Cycle in an Immortalized Human Foreskin Keratinocyte Cell Line

Virology ◽  
1999 ◽  
Vol 262 (2) ◽  
pp. 344-354 ◽  
Author(s):  
Elsa R. Flores ◽  
B.Lynn Allen-Hoffmann ◽  
Denis Lee ◽  
Carol A. Sattler ◽  
Paul F. Lambert
Keyword(s):  
Human Papillomavirus ◽  
Life Cycle ◽  
Cell Line ◽  
Hpv 16 ◽  
Human Papillomavirus Type 16 ◽  
Keratinocyte Cell

scholarly journals The Effects of Lycopene on the Methylation of the GSTP1 Promoter and Global Methylation in Prostatic Cancer Cell Lines PC3 and LNCaP

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Li-Juan Fu ◽  
Yu-Bin Ding ◽  
Lan-Xiang Wu ◽  
Chun-Jie Wen ◽  
Qiang Qu ◽  
...  
Keyword(s):  
Cell Line ◽  
Dna Methyltransferase ◽  
Suppressor Gene ◽  
In Vivo Studies ◽  
Global Methylation ◽  
Protein Levels ◽  
Long Interspersed Element ◽  
Androgen Independent

DNA (cytosine-5-) methylation silencing of GSTP1 function occurs in prostate adenocarcinoma (PCa). Previous studies have shown that there is an inverse relationship between dietary lycopene intake and the risk of PCa. However, it is unknown whether lycopene reactivates the tumor suppressor gene glutathioneS-transferase-π(GSTP1) by demethylation of the hypermethylated CpGs that act to silence the GSTP1 promoter. Here, we demonstrated that lycopene treatment significantly decreased the methylation levels of the GSTP1 promoter and increased the mRNA and protein levels of GSTP1 in an androgen-independent PC-3 cell line. In contrast, lycopene treatment did not demethylate the GSTP1 promoter or increase GSTP1 expression in the androgen-dependent LNCaP cell line. DNA methyltransferase (DNMT) 3A protein levels were downregulated in PC-3 cells following lycopene treatment; however, DNMT1 and DNMT3B levels were unchanged. Furthermore, the long interspersed element (LINE-1) and short interspersed element ALU were not demethylated when treated by lycopene. In LNCaP cells, lycopene treatment did not affect any detected DNMT protein expression, and the methylation levels of LINE-1 and ALU were decreased. These results indicated that the protective effect of lycopene on the prostate is different between androgen-dependent and androgen-independent derived PCa cells. Further, in vivo studies should be conducted to confirm these promising results and to evaluate the potential role of lycopene in the protection of the prostate.

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