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HPV18 E7 induces the over-transcription of eIF4E gene in cervical cancer.

Pang T, Wang S, Gao M, Kang H, Zhao Y, Yao Y, Hu X - Iran J Basic Med Sci (2015)

Bottom Line: Knock down of E7 decreased markedly eIF4E mRNA by 73% (P<0.01) in HPV18 positive HeLa cells.HPV E7 induced eIF4E gene over transcription which might be a new marker for CC.The finding broadens the understanding of the CC carcinogenesis.

View Article: PubMed Central - PubMed

Affiliation: Obstetrics and Gynecology Department of the first Affiliated Hospital of Guangdong Medical College, No. 57 Avenue of the people, Zhanjiang, Guangdong Province 524023, PR China ; Cancer Institute of Guangdong Medical College, No. 1 Xincheng Road, Dongguan, Guangdong Province 523808, PR China.

ABSTRACT

Objectives: Eukaryotic translation initiation factor 4E (eIF4E) is overexpressed in cervical cancer (CC). However, the molecular mechanisms are unclear. This study aimed to investigate the molecular mechanism of eIF4E gene overexpression in CC.

Materials and methods: The human papillomavirus (HPV) type 18 E7 and eIF4E mRNAs were measured following knock down or overexpression of E7 gene by RT-PCR and real-time PCR. Cell counting kit-8 assay was used to determine the cell proliferation. Flow cytometry was used to analyze the cell cycle and apoptosis. Transwell system was employed to determine the cell migration.

Results: Overexpression of E7 gene increased eIF4E mRNA level by 24.3% (P<0.01) in HPV negative C33A cells. Knock down of E7 decreased markedly eIF4E mRNA by 73% (P<0.01) in HPV18 positive HeLa cells. Under the state of high expression of E7, 1) up-regulation of eIF4E drastically promoted the cell proliferation, cell cycle and cell migration, and inhibited the cell apoptosis. 2) down-regulation of eIF4E significantly inhibited the cell proliferation, cell cycle and the ability of cell migration, and also promoted the apoptosis of cervical cancer cells.

Conclusion: HPV E7 induced eIF4E gene over transcription which might be a new marker for CC. The finding broadens the understanding of the CC carcinogenesis.

No MeSH data available.


Related in: MedlinePlus

Human papillomavirus e7 gene induced eukaryotic translation initiation factor 4e expression and promoted the proliferation and migration of C33A cells. Cells were divided into 4 groups: untreated C33A cell group (MOCK), p-EGFP blank plasmid group (NC); E7m, E7 mutant group; E7, E7 expression vector group. (A) ecto-E7 gene expression of C33A cells at 20 hr, detected by RT-PCR. (B) eIF4E gene expression of C33A cells at 20 hr, detected by RT-PCR. (C) Transfection of E7 gene promoted the proliferation of C33A cells, detected by CCK-8 assay. (D) Transfection of E7 gene promoted the migration of C33A cells, detected by the transwell migration assay. *: vs Mock, P <0.05; **: vs Mock, P<0.01
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Figure 1: Human papillomavirus e7 gene induced eukaryotic translation initiation factor 4e expression and promoted the proliferation and migration of C33A cells. Cells were divided into 4 groups: untreated C33A cell group (MOCK), p-EGFP blank plasmid group (NC); E7m, E7 mutant group; E7, E7 expression vector group. (A) ecto-E7 gene expression of C33A cells at 20 hr, detected by RT-PCR. (B) eIF4E gene expression of C33A cells at 20 hr, detected by RT-PCR. (C) Transfection of E7 gene promoted the proliferation of C33A cells, detected by CCK-8 assay. (D) Transfection of E7 gene promoted the migration of C33A cells, detected by the transwell migration assay. *: vs Mock, P <0.05; **: vs Mock, P<0.01

Mentions: The E7 expression vector was transfected into C33A cells. For RT-PCR results, two bands with expected sizes for E7 (271bp) (Figure 1A), E7 mutant (271bp) and GAPDH (224bp) were seen. The eIF4E mRNA detection showed two bands for eIF4E (132bp) and GAPDH (224bp) (Figure 1B). By Image J analysis, the relative eIF4E mRNA levels (eIF4E/GAPDH) were 0.682 (mock), 0.613 (NC), 0.808 (E7m group) and 0.762 (E7 group). The eIF4E bands were stronger in E7 and E7m groups than the mock and NC groups. The changes of the eIF4E mRNA were consistent with the changes of E7 mRNA. The lighter changes of the eIF4E mRNA in E7m group suggested that the E7m had a part of function in inducing eIF4E transcription. The results of real-time PCR were similar to that of RT-PCR.


HPV18 E7 induces the over-transcription of eIF4E gene in cervical cancer.

Pang T, Wang S, Gao M, Kang H, Zhao Y, Yao Y, Hu X - Iran J Basic Med Sci (2015)

Human papillomavirus e7 gene induced eukaryotic translation initiation factor 4e expression and promoted the proliferation and migration of C33A cells. Cells were divided into 4 groups: untreated C33A cell group (MOCK), p-EGFP blank plasmid group (NC); E7m, E7 mutant group; E7, E7 expression vector group. (A) ecto-E7 gene expression of C33A cells at 20 hr, detected by RT-PCR. (B) eIF4E gene expression of C33A cells at 20 hr, detected by RT-PCR. (C) Transfection of E7 gene promoted the proliferation of C33A cells, detected by CCK-8 assay. (D) Transfection of E7 gene promoted the migration of C33A cells, detected by the transwell migration assay. *: vs Mock, P <0.05; **: vs Mock, P<0.01
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4556762&req=5

Figure 1: Human papillomavirus e7 gene induced eukaryotic translation initiation factor 4e expression and promoted the proliferation and migration of C33A cells. Cells were divided into 4 groups: untreated C33A cell group (MOCK), p-EGFP blank plasmid group (NC); E7m, E7 mutant group; E7, E7 expression vector group. (A) ecto-E7 gene expression of C33A cells at 20 hr, detected by RT-PCR. (B) eIF4E gene expression of C33A cells at 20 hr, detected by RT-PCR. (C) Transfection of E7 gene promoted the proliferation of C33A cells, detected by CCK-8 assay. (D) Transfection of E7 gene promoted the migration of C33A cells, detected by the transwell migration assay. *: vs Mock, P <0.05; **: vs Mock, P<0.01
Mentions: The E7 expression vector was transfected into C33A cells. For RT-PCR results, two bands with expected sizes for E7 (271bp) (Figure 1A), E7 mutant (271bp) and GAPDH (224bp) were seen. The eIF4E mRNA detection showed two bands for eIF4E (132bp) and GAPDH (224bp) (Figure 1B). By Image J analysis, the relative eIF4E mRNA levels (eIF4E/GAPDH) were 0.682 (mock), 0.613 (NC), 0.808 (E7m group) and 0.762 (E7 group). The eIF4E bands were stronger in E7 and E7m groups than the mock and NC groups. The changes of the eIF4E mRNA were consistent with the changes of E7 mRNA. The lighter changes of the eIF4E mRNA in E7m group suggested that the E7m had a part of function in inducing eIF4E transcription. The results of real-time PCR were similar to that of RT-PCR.

Bottom Line: Knock down of E7 decreased markedly eIF4E mRNA by 73% (P<0.01) in HPV18 positive HeLa cells.HPV E7 induced eIF4E gene over transcription which might be a new marker for CC.The finding broadens the understanding of the CC carcinogenesis.

View Article: PubMed Central - PubMed

Affiliation: Obstetrics and Gynecology Department of the first Affiliated Hospital of Guangdong Medical College, No. 57 Avenue of the people, Zhanjiang, Guangdong Province 524023, PR China ; Cancer Institute of Guangdong Medical College, No. 1 Xincheng Road, Dongguan, Guangdong Province 523808, PR China.

ABSTRACT

Objectives: Eukaryotic translation initiation factor 4E (eIF4E) is overexpressed in cervical cancer (CC). However, the molecular mechanisms are unclear. This study aimed to investigate the molecular mechanism of eIF4E gene overexpression in CC.

Materials and methods: The human papillomavirus (HPV) type 18 E7 and eIF4E mRNAs were measured following knock down or overexpression of E7 gene by RT-PCR and real-time PCR. Cell counting kit-8 assay was used to determine the cell proliferation. Flow cytometry was used to analyze the cell cycle and apoptosis. Transwell system was employed to determine the cell migration.

Results: Overexpression of E7 gene increased eIF4E mRNA level by 24.3% (P<0.01) in HPV negative C33A cells. Knock down of E7 decreased markedly eIF4E mRNA by 73% (P<0.01) in HPV18 positive HeLa cells. Under the state of high expression of E7, 1) up-regulation of eIF4E drastically promoted the cell proliferation, cell cycle and cell migration, and inhibited the cell apoptosis. 2) down-regulation of eIF4E significantly inhibited the cell proliferation, cell cycle and the ability of cell migration, and also promoted the apoptosis of cervical cancer cells.

Conclusion: HPV E7 induced eIF4E gene over transcription which might be a new marker for CC. The finding broadens the understanding of the CC carcinogenesis.

No MeSH data available.


Related in: MedlinePlus